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The mycotoxin patulin is subjected to European Union statutory limits, and has action levels imposed in the United States of America in fruit products. Samples from soil and an orchard were analysed by primers of the isoepoxydon dehydrogenase gene of the patulin biosynthetic pathway as an HACCP procedure in an initial assessment. A non-orchard soil appeared to inhibit the reaction. However, the reaction was positive from orchard soils. Negative results from orchard samples appeared to be because of target DNA being absent. Bramley samples contained the gene frequently. The method may be useful in the determination of critical control points. ; Fundação para a Ciência e a Tecnologia (FCT); UK Food Standards Agency; CABI ...

International audience ; Patulin is a mycotoxin produced by species of Penicillium, Aspergillus and Byssochylamys. Several Scientific Committees classify patulin as mutagenic, embriotoxic and immunotoxic; it has been found as a natural contaminant of processed apple products and its presence may be indicative of the quality of fruit used in production. In this work a method for the analysis of patulin is described, this methodology is based on a simple liquid-liquid extraction with acetonitrile; patulin is analyzed using liquid chromatography with UV detection. Patulin identity was confirmed by GC-MS after its reaction with N-Methyl-N-(trimethylsilyl) trifluoroacetamide. Fifty-three apple-containing products were analyzed with the method described in this work. Patulin was detected in 14 of the samples in a range of 1.5-50.9 μg L-1; six of them were above the maximum permitted level by the European Union. Based on these results and juice consumption by the Spanish adult population, patulin estimated intake was 0.42 ng kg-1 body weight per day.

Interactions between fungi occur when they grow on the same host plant. This is the case of Botrytis cinerea and Penicillium expansum on grape. P. expansum is also responsible for production of the mycotoxin patulin. In this study, the influence of the interaction between both fungi on fungal growth parameters was studied as well as the effect on the accumulation of patulin by P. expansum. For that purpose, spores of B. cinerea and P. expansum were inoculated together (mixed inoculum), and the parameters growth rate, time for growth and patulin accumulation were assessed. The presence of P. expansum conidia shortened the time for growth of mixed inoculum colonies which, at the end of incubation, were B. cinerea-like. Although some P. expansum growth was observed in mixed inoculum colonies, very low levels of patulin were observed. In assays carried out in patulin-spiked medium, B. cinerea was capable to metabolize the mycotoxin. The capabilities of B. cinerea to shorten time for growth and prevent patulin accumulation are competing abilities that facilitate grape colonization. Significance and Impact of the Study: To our knowledge, this is the first study on the influence of fungal interactions between Botrytis cinerea and Penicillium expansum on growth parameters and patulin accumulation. The incidence of P. expansum in some wine regions is high, but the attack of this fungus to vineyards is rare, being B. cinerea the most common disease. In this assay, it was observed that, in vitro, the presence of P. expansum spores enhanced B. cinerea growth, while the latter avoided patulin accumulation. ; H. Morales is grateful to the Portuguese Government. (Ministerio da Ciencia, Tecnologia e Ensino Superior; FCT Fundacao para a Ciencia e a Tecnologia. Grant ref. ...

Wild yeasts isolated from the surface of apples were screened for antagonistic activity against Penicillium expansum, the main producer of the mycotoxin patulin. Three antagonistic yeasts (Y33, Y29 and Y24) from a total of 90 were found to inhibit P. expansum growth. Identification by ITS region sequence and characterization showed that three selected isolates of yeast should be different strains of Metschnikowia pulcherrima. Several concentrations of the selected yeasts were used to study their in vitro antifungal effectivity against P. expansum on Petri dishes (plates with 63.6 cm2 surface) whereas their potential activity on patulin reduction was studied in liquid medium. Finally, the BCA that had the best in vitro antifungal capacity against P. and the best patulin degradation capacity was selected to be assessed directly on apples. All the selected strains demonstrated antifungal activity in vitro but the most efficient was the strain Y29. Isolated strains were able to reduce patulin content in liquid medium, Y29 being the only strain that completely reduced patulin levels within 120 h. The application of Y29 as biocontrol agent on the surface of apples inoculated with P. expansum, inhibited fungal growth and patulin production during storage. Therefore, the results shown that this yeast strain could be used for the reduction of P. expansum and its mycotoxin in apples or apple-based products by adapting the procedure application. ; The authors acknowledge the financial support of the Agence Nationale de la Recherche (France) (PATRISK project: ANR-17-CE21-0008) and the financial support of the European Union (Erasmus grant attributed to Laura Settier-Ramírez). ; Peer reviewed

The European Union (EU) has introduced statutory limits for patulin in fruit products. Species definitions need to be unambiguous for this capability, especially to determine 'weak spots' in food, drink and feed production. Fungi were analysed for the isoepoxydon dehydrogenase (IDH) gene of the patulin metabolic pathway to indicate potential patulin production. In several cases inhibition of PCR product formation was indicated. Dilution as a means of overcoming PCR inhibition and to determine optimal concentrations was assessed and the ramifications for nucleic acid analysis in general are highlighted. The inhibitors may be secondary metabolites. However, inhibition was not involved obviously for Penicillium brevicompactum. The gene was detected frequently from Aspergillus section Clavati and Penicillium subgenus Penicillium species. Some strains within certain species were negative for the gene. Detection of the gene product in Byssochlamys is reported. ; UK Food Standards Agency - contract no. ...

WOS: 000322829700003 ; PubMed ID: 22988003 ; This study describes a capillary zone electrophoretic method for the determination of patulin. An optimum run buffer was found to be 25 mM of sodium tetraborate and 10% acetonitrile (v/v) at pH 10. Optimum conditions were determined to be: an applied voltage of 25 kV (normal polarity), temperature of 25 degrees C and injection time of 10 s at 50 mbar; the signals of patulin and phenobarbital as internal standard were detected at 276 nm. The method was highly reproducible, with relative standard deviations of 0.02-0.85 for intra-day and 0.04-0.42 for inter-day for standard patulin. Acceptable linearity [y = 0.0020C (mu g/L) - 0.0680 (r = 0.9999)] was obtained over a concentration range of 0.25 to 4.99 mu g/mL of patulin. The limits of detection and quantification were calculated to be 5.9 x 10(-3) and 1.79 x 10(-2) mu g/mL, respectively. Recovery was 68.0%. The proposed method was applied to 21 apple juice samples purchased from different Turkish markets, and two were found to contain higher than the limits of the European Union Directive for patulin. ; Eskisehir Osmangazi University Scientific Research Projects [200719037] ; The authors are grateful to Eskisehir Osmangazi University Scientific Research Projects (Project No. 200719037) for their financial support for this work, ChemAxon for access of Marvin Beans 5.7.0 program and Edward McQuaid for reviewing the final English draft.

This research aims to assess the natural occurrence of patulin (PAT) in selected citrus fruits from central cities of Punjab and Pakistan's northern cities. A total of 2970 fruit samples from 12 citrus cultivars were examined using liquid chromatography fitted with a UV detector. The detection limit (LOD) and quantification limit were 0.04 and 0.12 µg/kg, respectively. About 56% of samples of citrus fruits from Punjab's central cities, Pakistan, were found to be contaminated with PAT, with values ranging from 0.12 to 1150 µg/kg in samples from central Punjab cities. Furthermore, 31.7% of samples of citrus fruits from northern cities of Pakistan were contaminated with PAT, with values ranging from 0.12 to 320 µg/kg. About 22.1% of citrus fruit samples had PAT levels greater than the suggested limits established by the European Union (EU). The dietary intake levels of PAT ranged from 0.10 to 1.11 µg/kg bw/day in the central cities of Punjab, Pakistan, and 0.13 to 1.93 µg/kg bw/day in the northern cities of Pakistan.

Patulin is a water-soluble mycotoxin produced by several species of fungi. Governmental bodies have placed it under scrutiny for its potential negative health effects, and maximum residue limits are fixed in specific food matrices to protect consumers' health. Confirmatory analysis of patulin in complex food matrices can be a difficult task, and sample clean-up treatments are frequently necessary before instrumental analyses. With the aim of simplifying the clean-up step, we prepared a 256-member combinatorial polymeric library based on 16 functional monomers, four cross-linkers and four different porogenic solvents. The library was screened for the binding towards patulin in different media (acetonitrile and citrate buffer at pH 3.2), with the goal of identifying polymer formulations with good binding properties towards the target compound. As a proof of concept, a methacrylic acid-co-pentaerithrytole tetraacrylate polymer prepared in chloroform was successfully used as a solid-phase extraction material for the clean-up and extraction of patulin from apple juice. Clean chromatographic patterns and acceptable recoveries were obtained for juice spiked with patulin at concentration levels of 25 (64 ± 12%), 50 (83 ± 5.6%) and 100 μg L−1 (76 ± 4.5%). The within-day and between-day reproducibility evaluated at a concentration level of 25 μg L−1 were 5.6 and 7.6%, respectively.

Article ; Penicillium expansum infection of apples and mycotoxin patulin (PAT) production has previously been associated with many pre- and postharvest factors other than physiological disorders. In the current study, 'Antei' and 'Krameri tuviõun' apples with and without bitter pit (BP) symptoms and 'Talvenauding' apples with and without superficial scald (SS) symptoms were used in order to determine if the named physiological disorders may influence susceptibility to P. expansum infection and PAT production. Apples were inoculated with 10 μL P. expansum spore suspension with the concentration of 1×105 conidia mL-1 and stored at 24 °C with relative humidity (RH) 80%. After 7 and 11 days, lesion diameters were measured, and apples were pressed into juice. PAT content was determined in pasteurized juice. Two cultivars out of three showed that in fruit with physiological disorders, Penicilllium infection and PAT production proceeded significantly faster compared to apples, which did not have physiological disorders. SS increased the risk for PAT occurrence in juice more than BP: while the juice pressed from BP–affected apples with no visual signs of fungal diseases did not contain PAT, juice pressed from apples with SS contained PAT three times above legislative limits defined by the World Health Organization (50 μg L-1).