ABSTRACT
OBJECTIVE To investigate the toxicological effect of patulin(PAT)on the growth of human normal liver cells L-02 and its possible mechanisms. METHODS After cells were treated with PAT 1.25, 2.5,5,10 and 20μmol·L-1 for 24 or 48 h,cell viability was examined using MTT assay. L-02 cells were treated with PAT 5 and 10 μmol · L- 1 for 24 h ,respectively. Cytomorphology and mitochondrial membrane potential (MMP) were observed under a fluorescence microscope. Apoptosis,MMP and reactive oxygen species (ROS)were analyzed by flow cytometry. Mitochondria apoptosis pathways were detected by Western blotting. RESULTS PAT exhibited a strong inhibitory effect on L-02 in a concentration-dependent and time-dependent manner. IC50 of PAT treatment for 24 or 48 h was 6.61 and 2.78 μmol · L-1,respectively. MMP was decreased,while the percentage of low MMP cells increased from(9.2±2.3)%in controls to(23.4±4.5)%( PAT 5μmol·L-1)and(47.1±5.5)%(PAT 10μmol·L-1), respectively. Compared to untreated cells,the early apoptosis population increased from(3.8±1.1)%to(29.8±4.5)%( PAT 5μmol·L-1)and (24.1±6.2)%(PAT 10μmol·L-1)(P<0.01),respectively. Further?more,the accumulation of ROS was also observed. The effect of PAT on ROS and cell viabilities could be attenuated by glutathione. CONCLUSION PAT can significantly inhibit the growth of L-02 and induce apoptosis via ROS-dependent mitochondria pathways.
ABSTRACT
OBJECTIVE: To characterize the antibacterial agents from the culture broth of Penicillium sp. I09F 484. METHODS: The compounds were purified by a combination of chromatographic methods, and their structures were identified by IR, MS, CD, 1D and 2D-NMR spectroscopic data analysis. The antimicrobial activities of the isolates were evaluated by the micro-broth dilution method and the antiviral activities were also assessed. RESULTS: Eight compounds were identified from the cultures of strain I09F 484. Their structures were elucidated as patulin(1), gentisyl alcohol(2), 3-hydroxybenzyl alcohol(3), 2-methylhydroquinone(4), (45, 55)-4, 5-dihydroxy-2-methylcyclohex-2-enone(5), epiepoformin(6), thymidine(7), cyclo(L-Pro-L-Tyr) (8). Compound 1 displayed moderate antibacterial activities with MICs ranging 64-128 μg · mL-1 against a number of pathogenic strains, while 4 was active against Staphylococcus aureus and Staphylococcus epidermidis. Compound 2 displayed antiviral activity against influenza A strains A/FM/1/47 (H1N1) nd A/Hanfang/359/95( H3N2) viruses, with IC50 values of 5.0 and 5.9 μmol · L-1 Respectively. Compound 4 inhibited Coxsackie virus B3 replication, with an IC50 value of 149.4 μmol · L-1. CONCLUSION: Compounds 1-5 are obtained from the strain for the first time. The antibacterial activity of the crude extracts is mainly due to the presence of the known patulin (1). Compounds 2 and 4 show moderate influenza and Coxsackie virus B3 inhibitory activity and the antiviral activities are first assessed.
ABSTRACT
Objective:To establish an HPLC method for the determination of patulin in health foods containing hawthorn to im-prove the quality control of patulin in related health foods. Methods:An Agilent TC-C18(2)(250 mm ×4.6 mm, 5 μm) column was used with the temperature of 30℃. The mobile phase consisted of 0. 8% tetrahydrofuran and the flow rate was 1. 0 ml·min-1 . The detection wavelength was set at 276nm. Results:The calibration curve of patulin was linear within the range of 1-20ng(r=1. 000 0), the average recovery was 92. 1% and RSD was 2. 2%(n=6). Conclusion:The method is simple, reliable and accurate, which can be used in the content determination of patulin in health foods containing hawthorn.
ABSTRACT
A part of apples destined to juice production is generally of poor quality. Apples from cold storage or recently harvest (ground harvested or low quality apples) are stored under ambient conditions until they are processed. Since Penicillium expansum and P. griseofulvum are the principal fungal species isolated from stored apples in Brazil, the objective of this study was to investigate the ability of these strains to produce patulin in apples and report the consequences of this type of storage in loss of quality. The toxin was quantified using thin layer chromatography and charge-coupled device camera (TLC-CCD). The rate and quantities that P. expansum and P. griseofulvum can grow and produce patulin are highly dependent on the fungal strain and time. Lesion diameter resulted to be independent of the strain considered. The maximum period of time which apples were kept at cold storage (4 ºC) without patulin accumulation was 27 days. When these apples were kept at 25 ºC during 3 days, both factors lesion diameter and patulin production increased significantly. These results confirm that time in which apples are taken out from cold storage room before juice production is critical in order to prevent patulin accumulation.
Subject(s)
Food Preservation , Food Storage , Malus , Mycotoxins/analysis , Patulin/analysis , Penicillium/isolation & purification , Chromatography, Thin Layer , Food Samples , Carbonated Beverages , MethodsABSTRACT
A patulina é uma micotoxina produzida por várias espécies de Penicillium, Aspergillus e Byssochlamys. Em experimentos com animais, ela demonstrou ter atividade mutagênica, carcinogênica e teratogênica. Tem sido freqüentemente encontrada em maçãs e derivados. A patulina é facilmente transferida da maçã para o suco durante o processamento devido a sua alta solubilidade em água. Essa micotoxina é muito estável ao aquecimento em meio ácido, como no suco de maçã. Assim, a presença de patulina em suco de maçã é um indicador da qualidade das maçãs utilizadas no processamento. Muitos métodos têm sido desenvolvidos para a determinação da patulina, principalmente baseados na extração líquido-líquido com acetato de etila e determinação por CLAE. É importante evidenciar a necessidade de legislação que regulamente limites dessa micotoxina em alimentos no Brasil. Esta revisão bibliográfica tem como objetivos descrever as principais características da patulina, a ocorrência, os aspectos toxicológicose os métodos desenvolvidos para sua detecção e controle durante os estágios da produção da maçã e suco.
Patulin is a mycotoxin produced by several Penicillium, Aspergillus and Byssochlamys species. Patulin is a highly toxic compound which has shown to be mutagenic, carcinogenic and teratogenic in experiments with animals. It has often been found in apples and apple products. Patulin is easily transfered into apple juice during processing due to its high solubility in water. This mycotoxin is very stable to heat in acidic medium as in apple juice. Thus, patulin content of apple juice is an indicator of the quality of the apples used to juice production. Many methods have been developed for the patulin determination mainly based on liquid-liquid extraction with ethyl acetate and use of HPLC for detection. It is important to show the need of legislation that imposes patulin limits in foods in Brazil. The objectives of this review are to describe the main patulin characteristics, occurrence, toxicological aspects, methods developed for patulin detection and control during the stages of apple and juice production.
ABSTRACT
A podridão ácida é uma patologia das videiras, de alta freqüência nos períodos quentes e chuvosos, em conseqüência do desenvolvimento de fungos e bactérias em seus frutos. Um dos gêneros de fungos responsáveis por esta doença é o Penicillium spp., que poderá levar à produção de micotoxinas, como a patulina. Neste trabalho, foi realizada uma investigação sobre a podridão ácida e a ocorrência de patulina em uvas Itália (Vitis vinifera L. cv. Rubi) cultivadas no município de Marialva, Brasil. Foram avaliados os seguintes tratamentos: (I) testemunha sem sintomas de podridão ácida; (II) amostras com sintomas, sem a remoção das bagas infectadas; (III) amostras com sintomas, com a remoção das bagas infectadas, (IV) amostras com sintomas da doença, com a remoção das bagas infectadas e desinfecção com hipoclorito. A cromatografia em camada delgada foi utilizada para a determinação de patulina. Nas amostras analisadas, não foi detectada contaminação por esta micotoxina.
Sour rout is a pathology of bunch grapes occurring in hot and rainy periods. Several fungi and bacteria are associated with this disease, including Penicilium spp. In the present study it was carried out an investigation to determine the incidence of sour rout and patulin in grapes (Vitis vinifera L. cv. Rubi) cultivated in Marialva, Brazil. The treatments studied were the following: (I) samples without symptoms of sour rout, (II) samples with symptoms and without cleaning stage, (III) samples with symptoms and with cleaning stage and (IV) samples with symptoms, cleaning stage and washing with hypochlorite. The thin-layer chromatography was utilized to determine the presence and quantification of patulin. Patulin was not detected in any samples.
ABSTRACT
A Região Sul do Brasil é grande produtora de maçã, sendo 80 por cento destinada ao consumo in natura. As micotoxinas são metabólitos secundários de fungos presentes na cadeia alimentar como contaminantes, causando diversos efeitos toxicológicos e imunológicos. Considerando que a patulina seja uma micotoxina produzida por Penicillium expansum, principal contaminante da maçã propôs-se investigar a ação de extratos aquoso e oleoso de Nim (Azadirachta indica) em maçã artificialmente contaminada. Foram testados dois tipos de extratos: o extrato aquoso obtido de maceração de folhas de Nim nas concentrações de 5, 10, 20 e 30 por cento, e o extrato oleoso comercial (DalNeem®), obtido de sementes de Nim, nas concentrações de 0,125, 0,25, 0,5, 1, 2 e 5 por cento. Os extratos aquosos não alteram a produção da patulina em maçãs contaminadas artificialmente por P. expansum, mas o extrato oleoso obtido das sementes da planta diminuiu acentuadamente a produção de patulina, inclusive em concentrações inferiores a 0,5 por cento.
The South region of Brazil is a major apple producer, where 80 percent is destined to the "in natura" consumption. The mycotoxins are fungal secondary metabolites present as contaminants in the food chain, causing several toxicological and immunological effect. Patulin is a mycotoxin mainly produced by Penicillium expansum and well known as the main contaminant in apples. Due to this contamination, the aim of this work was to access the effect of Neem extract (Azadirachta indica) on artificially contaminated apples. Two types of Neem extracts were tested: the aqueous extracts of Neem leafs at 5, 10, 20 and 30 percent concentrations, and commercial seed oil of Neem (DalNeem®) at 0.125, 0.25, 0.5, 1, 2 and 5 percent in water. Although the addition of Neem aqueous extracts in artificially contaminated apples with P. expansum was unable to affect the patulin production, the Neem seed oil extract at concentrations as lower as 0.5 percent, caused a pronounced diminution over patulin production.
ABSTRACT
Taking into account the preliminary antagonistic/biodegradation property showed by Pichia membranifaciens and Sporobolomyces roseus, which decreased the initial patulin concentration of 588.4 to 290.0 mug/mL, ability of P. ohmeri 158 in biocontrol against Penicillium expansum and patulin decrease in vitro was performed. The culture supernatant of P. ohmeri 158 was effective against 66.17 percent micelial growth, indicating antibiosis related with the killer phenomenon. The initial patulin concentration of 223 mug in the presence of P. ohmeri 158 cells was decreased over 83 percent of the original concentration, when incubated at 25°C/2 days and > 99 percent after 5 days incubation time, with undetectable patulin level after 15 days. The initial pH 4.0 decreased to pH 3.3 along 15 days experiment, suggesting that patulin decrease was an active process and a consequence of yeast metabolism. The results suggested that P. ohmeri 158 could be a promising alternative for the inhibition of P. expansum growth and patulin degradation.
Considerando o antagonismo e degradação de patulina detectados em Pichia membranifaciens e Sporobolomyces roseus no estudo preliminar, este trabalho avaliou o efeito antagônico de Pichia ohmeri 158 no desenvolvimento de Penicillium expansum e a degradação de patulina "in vitro". O sobrenadante do cultivo de P. ohmeri 158 inibiu 66,17 por cento do desenvolvimento micelial, indicando antibiose relacionada ao fator killer. A concentração inicial de patulina (223 mug) na presença de células íntegras de P. ohmeri foi reduzida em mais de 83 por cento após dois dias de incubação a 25°C e superior a 99 por cento após 5 dias, com níveis indetectáveis no 15° dia. O decréscimo do pH 4,0 inicial para pH 3,3 sugeriu que a eliminação de patulina é um processo ativo e uma conseqüência do metabolismo da levedura. Os resultados obtidos concluem que P. ohmeri 158 é uma alternativa promissora na inibição do desenvolvimento de P. expansum e na degradação de patulina.
Subject(s)
Antibiosis , Food Technology , Patulin , Pest Control, Biological , PichiaABSTRACT
Patulina é uma micotoxina produzida por várias espécies de fungos (Penicillium, Aspergillus e Biossochlamys), principalmente pelo Peniccillium expansum, fungo presente nas maçäs podres. É um composto tóxico, que em experimentos com animais tem demonstrado ter atividade mutagênica, carcinogênica. O conteúdo de patulina em suco de maçä é um bom indicador da qualidade das maçäs utlizadas no processamento. Treze marcas comercializadas em Belo Horizonte/Minas Gerais foram coletadas entre março e setembro de 1999. A metodologia utilizada foi a cromatografia líquida de alta eficiência (CLAE) em fase reversa. Patulina foi extraída com acetato de etila e o extrato foi purificado por participaçäo com soluçäo de carbonato de sódio. A separaçäo e quantificaçäo foi realizada em um cromatógrafo líquido por meio de uma coluna de fase reversa C18(3um) e detetor de ultravioleta a 278nm. Patulina e 5-hidroximetilfurfural (5-HMF) foram completamente separados utilizando como fase móvel 0,8(por cento) tetrahidrofurano em água contendo 0,02(por cento) de azida de sódio em uma vazäo de 0,5mL/minuto. O limite de detecçäo foi <5ug/L. Foram obtidos recuperaçöes média de patulina superiores a 73(por cento) e coeficientes de variaçäo menores que 7(por cento). Patulina foi detectada em apenas uma amostra (10ug/L). Apesar da baixa incidência de patulina em suco de maçä neste estudo, autoridades brasileiras de saúde devem continuar a monitorar estes produtos. (AU)
Patulin is a mycotoxin produced by several species of moulds (Penicillium, Aspergillus andBissochlamys), but mainly by the apple-rotting fungus Penicillium expansum. Patulin is a highly toxiccompound that has been shown to be mutagenic, carcinogenic and teratogenic in experiments with animals.The patulin content of apple juice is a good indicator of the quality of the apples used in the process.Thirteen brands were collected from retail markets in the city of Belo Horizonte, state of Minas Gerais,Brazil, between March and September 1999. A rapid reversed-phase liquid chromatographic determinationof patulin was used. Patulin was extracted with ethyl acetate and the extract was cleaned up by partitionwith sodium carbonate solution. The identification and quantification of patulin was determined by C18reversed phase column (3 µm) and a variable-wavelength UV-VIS detector set at 278 nm. Patulin and 5-hydroxymethylfurfural were completely resolved by using 0,8% tetrahydrofuran in water containing 0,02%sodium azide as the mobile phase at a flow rate of 0,5 mL/min. The detection limit was < 5 µg/L. Averagerecoveries greater than 73% of patulin and coefficients of variation of less than 7% were obtained. Patulinwas detected in only one sample (10 µg/L). Although the incidence of patulin in apple juice in the presentstudy was low, Brazilian health authorities should continue monitoring these products. (AU)