ABSTRACT
Clitoria ternatea or Commonly known blue pea, is a perennial climber crop native to Asian countries. The current study was aimed to evaluate the antimicrobial activity C. ternatea extract on food borne microorganisms and its antifungal effect on Penicillium expansum. The extract showed significant antimicrobial activity against 3 Gram positive bacteria, 2 Gram negative bacteria and 1 filamentous fungus on disc diffusion assay. The extract also showed good biocidal effect on all Gram positive bacteria tested and P. expansum. However, the kill curve analysis revealed that the fungicidal activity of the extract against P. expansum conidia was depend on the concentration of the extract and the time of exposure of the conidia to the extract. The scanning electron micrograph of the extract treated P. expansum culture showed alterations in the morphology of fungal hyphae. The germination of P. expansum conidia was completely inhibited and conidial development was totally suppressed by the extract, suggesting the possible mode of action of anthocyanin. Besides, the extract also exhibited 5.0-log suppression of microbial growth relative to control in the rice model. The results indicate the potential use of the C. ternatea anthocyanin as food biopreservative.
Subject(s)
Humans , Anthocyanins , Asian People , Clitoria , Diffusion , Fungi , Germination , Gram-Negative Bacteria , Gram-Positive Bacteria , Hyphae , Peas , Penicillium , Spores, FungalABSTRACT
Bacteria and molds may spoil and/or contaminate apple juice either by direct microbial action or indirectly by the uptake of metabolites as off-flavours and toxins. Some of these microorganisms and/or metabolites may remain in the food even after extensive procedures. This study aim to identify the presence of molds (including heat resistant species) and Alicyclobacillus spp., during concentrated apple juice processing. Molds were isolated at different steps and then identified by their macroscopic and microscopic characteristics after cultivation on standard media at 5, 25 and 37ºC, during 7 days. Among the 19 isolated found, 63% were identified as Penicillium with 50% belonging to the P. expansum specie. With regards to heat resistant molds, the species Neosartorya fischeri, Byssochlamys fulva and also the genus Eupenicillium sp., Talaromyces sp. and Eurotium sp. were isolated. The thermoacidophilic spore-forming bacteria were identified as A. acidoterrestris by a further investigation based on 16S rRNA sequence similarity. The large contamination found indicates the need for methods to eliminate or prevent the presence of these microorganisms in the processing plants in order to avoid both spoilage of apple juice and toxin production.
Subject(s)
Alicyclobacillus/isolation & purification , Beverages/microbiology , Food Handling , Fungi/classification , Fungi/isolation & purification , Bacterial Load , Colony Count, Microbial , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Malus , Microscopy , /genetics , Sequence Analysis, DNAABSTRACT
Ten Penicillium sp. were screened for lectin activity for occurrence of lectins. Mycelial extracts from submerged cultures of P. corylophilum, P. expansum and P. purpurogenum showed agglutination against human (A, B, AB and O), goat, sheep, pig and rabbit erythrocytes. Neuraminidase treatment to human blood type O erythrocytes substantially increased their agglutinability by all the lectins as compared to untreated erythrocytes. Modification of erythrocyte surfaces by protease increased the lectin titre only of P. corylophilum with no effect on other two lectins. P. corylophilum and P. expansum displayed relatively lower titres in mycelial extracts prepared from agar plate cultures as compared to broth cultures. A panel of sugars was tested for inhibition of lectin activity. All the lectins were found to be specific for asialofetuin, bovine submaxillary mucin, porcine stomach mucin, chondroitin-6-sulphate, D-sucrose and D-glucose. P. corylophilum lectin was expressed (Titre 8) by 5 day old cultures, reaching its maximum level (Titre 32) upon 8 days of cultivation, thereafter declin in lectin activity was observed. P. purpurogenum lectin was expressed by 7-10 days old cultures, while in P. expansum maximum lectin activity was elaborated by 5-8 days old cultures. Lectin extracts from all the three species were found to possess antimicrobial activities. Lectin extracts from the three Penicillium species displayed antifungal activity and antibacterial activity against Gram-negative and Gram-positive bacterial strains.
Subject(s)
Anti-Infective Agents/pharmacology , Hemagglutination Tests , Lectins/pharmacology , Microbial Sensitivity Tests , Penicillium/chemistry , Penicillium/classification , Species SpecificityABSTRACT
O bolor azul, causado por Penicillium expansum, é responsável por grandes perdas nos frutos de maçã armazenados. Buscando medidas alternativas aos tratamentos convencionais, avaliou-se o efeito de ácidos fenólicos (cinâmico, ferúlico, gálico, salicílico e vanílico) sobre Penicillium expansum e sobre a podridão ocasionada nos frutos. Em lâminas de microscopia escavadas, foram avaliados a porcentagem de germinação e o comprimento do tubo germinativo do fungo, empregando-se os fenólicos a 1, 2,5 e 5 mM. In vivo, avaliou-se a intensidade da podridão de Penicillium a partir da imersão dos frutos em soluções fenólicas a 2,5, 5 e 10 mM. Em todos os experimentos, o delineamento utilizado foi o inteiramente casualizado com 4 repetições, sendo a parcela experimental representada por uma gota em lâmina escavada (bioensaios in vitro) ou 4 frutos no interior de uma bandeja plástica (bioensaios in vivo). Dentre os fenólicos, os ácidos cinâmico e salicílico inibiram completamente a germinação do fungo a 2,5 mM, mas somente o ácido salicílico controlou a doença causada por Penicillium expansum quando misturado à suspensão de esporos, reduzindo a incidência do bolor azul em 80%. Desta forma o ácido salicílico apresenta potencial para o tratamento fitossanitário de frutos de maçã em pós-colheita.
The blue mold, caused by Penicillium expansum, is responsible for great losses in stored apple fruit. Searching alternatives measures to conventional treatment, the effect of phenolic acids (cinnamic, ferulic, gallic, vanillic and salicylic acid) on Penicillium expansum and on blue mold intensity were evaluated. In vitro, the percentage of germination and the germ tube length of the fungus were assessed, using the phenolics at 1, 2,5 and 5 mM. In vivo, we evaluated the severity of Penicillium decay in fruits immersed on phenolic solutions at 2,5, 5 and 10 mM. In all experiments, the experimental design was completely randomized with four replications, and the experimental plot was represented by a drop on a microscope slide cavity or by 4 fruits inside a plastic tray. Cinnamic and salicylic acids completely inhibited the germination of the fungus at 2,5 mM, but only salicylic acid controlled the disease caused by Penicillium expansum when it was mixed with a spore suspension, reducing the incidence of blue mold by 80%. Thus salicylic acid has potential to control post-harvest decays.
Subject(s)
Penicillium , Salicylic Acid , Phenolic Compounds , Fruit , FungiABSTRACT
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 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.