Contamination status and exposure risk of mycotoxins in Coicis Semen / 中国中药杂志

By investigating the contamination status and predicting the exposure risk of mycotoxin in Coicis Semen, we aim to provide guidance for the safety supervision of Chinese medicinal materials and the formulation(revision) of mycotoxin limit standards. The content of 14 mycotoxins in the 100 Coicis Semen samples collected from five major markets of Chinese medicinal materials in China was determined by UPLC-MS/MS. The probability evaluation model based on Monte Carlo simulation method was established after Chi-square test and One-way ANOVA of the sample contamination data. Health risk assessment was performed on the basis of margin of exposure(MOE) and margin of safety(MOS). The results showed that zearalenone(ZEN), aflatoxin B_1(AFB_1), deoxynivalenol(DON), sterigmatocystin(ST), and aflatoxin B_2(AFB_2) in the Coicis Semen samples had the detection rates of 84%, 75%, 36%, 19%, and 18%, and the mean contamination levels of 117.42, 4.78, 61.16, 6.61, and 2.13 μg·kg~(-1), respectively. According to the limit standards in the Chinese Pharmacopoeia(2020 edition), AFB_1, AFs and ZEN exceeded the standards to certain extents, with the over-standard rates of 12.0%, 9.0%, and 6.0%, respectively. The exposure risks of Coicis Semen to AFB_1, AFB2, ST, DON, and ZEN were low, while 86% of the samples were contaminated with two or more toxins, which needs more attention. It is suggested that the research on the combined toxicity of different mycotoxins should be strengthened to accelerate the cumulative exposure assessment of mixed contaminations and the formulation(revision) of toxin limit standards.

Determination of 10 mycotoxins in Hippophae Fructus medicinal and edible products by ultra-performance liquid chromatography- tandem mass spectrometry / 中国中药杂志

An analytical method for 10 mycotoxins in Hippophae Fructus medicinal and edible products was established in this study, and the contamination of their mycotoxins was analyzed. First of all, the mixed reference solution of ten mycotoxins such as aflatoxin, ochratoxin, zearalenone, and dexoynivalenol was selected as the control, and the Hippophae Fructus medicinal and edible products were prepared. Secondly, based on the ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) technology, 10 mycotoxins in Hippophae Fructus medicinal and edible products were quantitatively investigated and their content was determined. Finally, the contamination of mycotoxins was analyzed and evaluated. The optimal analysis conditions were determined, and the methodological inspection results showed that the 10 mycotoxins established a good linear relationship(r>0.99). The method had good repeatability, test sample specificity, stability, and instrument precision. The average recovery rates of 10 mycotoxins in Hippophae Fructus medicinal products, edible solids, and edible liquids were 90.31%-109.4%, 87.86%-107.8%, and 85.61%-109.1%, respectively. Relative standard deviation(RSD) values were 0.22%-10%, 0.75%-13%, and 0.84%-8.5%, repsectively. Based on UPLC-MS/MS technology, the simultaneous determination method for the limits of 10 mycotoxins established in this study has fast detection speed, less matrix interference, high sensitivity, and accurate results, which is suitable for the limit examination of 10 mycoto-xins in Hippophae Fructus medicinal and edible products.

Methods for synchronous detection of 14 mycotoxins in Pseudostellariae Radix and investigation on its contamination / 中国中药杂志

This study aimed to establish a method for synchronous detection of 14 mycotoxins in Pseudostellariae Radix and investigate its contamination with mycotoxins, so as to provide technical guidance for monitoring the quality of Chinese medicinal materials and medication safety. The sample was extracted with 80% acetonitrile in an oscillator for 1 h, purified using the modified QuEChERS purifying agent(0.1 g PSA + 0.3 g C_(18) + 0.3 g MgSO_4), and separated on a Waters HSS T3 chromatographic column(2.1 mm×100 mm, 1.8 μm). The gradient elution was carried out with 0.1% formic acid in water and acetonitrile, followed by the scanning in the multi-reaction monitoring(MRM) mode and the analysis of mycotoxin contamination in 26 Pseudostellariae Radix samples. The recovery rates of the established method were within the range of 82.17%-113.6%, with the RSD values less than 7% and the limits of quantification(LOQ) being 0.019-0.976 μg·kg~(-1). The detection rate of 14 mycotoxins in 26 batches of medicinal materials was 53.85%. The detection rate of sterigmatocystin(ST) was the highest, followed by those of zearalenone(ZEN), aflatoxin G_2(AFG_2), fumonisin B_1(FB_1), HT-2 toxin, and nivalenol(NIV). Their respective detection rates were 38.46%, 26.92%, 23.08%, 11.54%, 11.54%, and 7.69%, with the pollution ranges being 1.48-69.65, 0.11-31.05, 0.11-0.66, 0.28-0.83, 20.86-42.56, and 0.46-1.84 μg·kg~(-1), respectively. The established method for the detection of 14 mycotoxins is accurate, fast and reliable. The research results have very important practical significance for guiding the monitoring and prevention and control of exogenous fungal contamination of Chinese medicinal materials.

Investigation and research on fungal diversity of Platycladi Semen on Chinese market / 中国中药杂志

In the process of harvesting, production and processing, storage, and transportation, the traditional Chinese medicine Platycladi Semen is prone to mildew due to its own and environmental factors, which can nourish the production of toxic or pathogenic fungi, and even produce mycotoxins, which affects the safety of clinical medication. The 2020 edition of Chinese Pharmacopoeia limits the highest standard of aflatoxin content in Platycladi Semen. However, there are few studies on the fungal contamination of Platycladi Semen, and it is difficult to prevent and control it in a targeted manner. Therefore, based on the Illumina NovaSeq6000 platform, this article uses ITS sequence amplicon technology to analyze the distribution and diversity of fungi in 27 batches of commercially available Platycladi Semen in the Chinese market. A total of 10 phyla, 35 classes, 93 orders, 193 families, 336 genera, and 372 species of fungi were identified in China. Among them, Aspergillus, Alternaria spp. were dominant, 20 batches of samples were detected for A. flavus, 10 batches of samples were detected for A. nidulans, and all samples were detected for potential pathogenic fungi such as A. fumigatus and A. niger. According to diversity analysis, the diversity of the fungal communities in the samples from Gansu province was high, the samples in Shandong province contain the largest number of fungal species, and the samples in Guangxi province had the lo-west diversity and the least number of species. In most samples, pathogenic fungi such as A. fumigatus, A. niger, A. flavus, A. parasiticus were detected in varying degrees. This study systematically investigated the fungal contamination of Platycladi Semen from the markets in the last link of the its industrial chain, and clarified the distribution of Platycladi Semen fungi, especially toxin-producing fungi, and provided theoretical basis for the targeted prevention and control of fungal contamination in Platycladi Semen.

Influência da qualidade micotoxicológica e nutricional de híbridos de milho no custo da ração de frangos de corte / Influence of mycotoxicological and nutritional quality of maize hybrids on broiler chickens feed cost

Objetivou-se avaliar as variáveis micotoxicológicas e nutricionais de híbridos de milho com diferentes características que influenciam no custo da ração para frangos de corte. Foram avaliados 26 híbridos de milho geneticamente modificados nas safrinhas de 2016 e 2017, com diferentes germoplasmas, textura de endosperma e duração do ciclo. Nos híbridos, foram avaliados grãos avariados, fumonisinas (B1+B2) (FUM), aflatoxinas (B1+B2+G1+G2) (AFLA), zearalenona (ZEA), deoxinivalenol (DON), umidade, proteína bruta (PB), energia metabolizável aparente corrigida para balanço de nitrogênio (EMAn), aminoácidos digestíveis para aves (lisina, metionina, cistina e treonina) e o respectivo custo da ração inicial para frangos de corte, que foi calculada pelo custo mínimo. A prevalência de FUM, AFLA, ZEA e DON foi de 90, 17, 33 e 0%, com médias de 3067, 1, 38 e 0µg/kg nos dois anos, respectivamente. A média de EMAn e PB foi de 3264kcal/kg e 8,02%, respectivamente, e diferiu (P<0,05) nos dois anos. O custo da ração foi influenciado significativamente (P<0,05) por FUM, PB, EMAn nos dois anos. Híbridos com tecnologia Viptera apresentam menor concentração por FUM e menor custo da ração. Híbridos de ciclo precoce têm menor concentração de FUM, maiores percentuais de PB e de aminoácidos digestíveis e menor custo da ração.(AU)

The objective of this study was to evaluate the mycotoxicological and nutritional variables of maize hybrids with different characteristics that influence the broiler chicken's feed costs. In 2016 and 2017 winter crops, 26 genetically modified hybrids of maize with different germplasm, endosperm texture and cycle duration were evaluated. The analyzed variables were damaged grains, fumonisins (B 1 +B 2 ) (FUM), aflatoxins (B 1 +B 2 +G 1 +G 2 ) (AFLA), zearalenone (ZEA), deoxynivalenol (DON), moisture, crude protein (CP), apparent metabolizable energy corrected for nitrogen balance (AMEn), digestible amino acids for poultry (lysine, methionine, cystine and threonine) and the respective cost of the initial feed for broiler chickens calculated at the minimum cost. The prevalence of FUM, AFLA, ZEA and DON was 90, 17, 33 and 0%, with means of 3067, 1, 38 and 0µg/kg in the two years, respectively. The mean of AMEn and CP was 3264kcal/kg and 8.02%, respectively, and differed (P< 0.05) in the two years. The feed cost was significantly influenced (P<0.05) by FUM, PB, AMEn in two years. Hybrids with Viptera technology show lower concentration per FUM and lower feed cost. Early cycle hybrids have lower concentrations of FUM, higher percentages of CP and digestible amino acids, and lower feed costs.(AU)

Biogenic amines and mycotoxins concentrations in baled silage from organic and conventional farms / Concentrações de aminas biogênicas e micotoxinas em silagem embalada de fazendas orgânicas e convencionais

The aim of the current study was to investigate biogenic amines and mycotoxins concentrations in baled silage (mainly Poaceae family grasses) prepared in organic and conventional farms and to relate these parameters to fermentative parameters. The mean dry matter (DM) content was 364.10±93.31 and 424.70±95.93g/kg in the silage from organic and conventional farms respectively. The silage samples from organic farms had 17.00% higher (P≤ 0.05) tyramine (TY) than the silage from conventional farms. Conventional farm samples were characterized by 46.00% higher histamine (HIS) (P≤ 0.05), 9.80% higher putrescine (PUT) (P≤ 0.05), 17.30% higher cadaverine (CAD) (P≤ 0.05). Aflatoxins (AFL) (total) and zearalenone (ZEN), T-2/HT-2 concentrations were higher respectively 16.00% (P≤ 0.05) and 13.40% (P≤ 0.05), 1.80% (P≤ 0.05) in the silage prepared in organic farms. Deoxynivalenol (DON) concentration was higher 42.40% (P≤ 0.05) in silage from conventional farms. Volatile fatty acids (VFA), lactic acid, ethanol, pH and ammonia nitrogen showed that the silage samples from organic and conventional farms were of good quality. Our study suggests differences in biogenic amine formation or mycotoxins content in silage from organic and conventional farming, but, overall, the measured values are too low to be relevant for animal health. Furthermore, these differences might as well be due to the difference in dry matter content and plant maturity between the organic and conventional silage samples.(AU)

O objetivo do presente estudo foi investigar concentrações de aminas biogênicas micotoxinas em silagem embalada (principalmente gramíneas da família Poaceae) preparada em fazendas orgânicas e convencionais e relacionar esses parâmetros a parâmetros fermentativos. A massa seca média (MS) foi 364,10±93,31 e 424,70±95,93g/kg na silagem de fazendas orgânicas e convencionais, respectivamente. As amostras de silagem de fazendas orgânicas tinham 17% a mais de tyramina (TY) (p≤ 0,05) que as de fazendas convencionais. As amostras de fazendas convencionais foram caracterizadas por histamina (HIS) 46,00% mais alta (P≤ 0,05), 9,80% putrecina (PUT) mais alta (P≤ 0,05), 17,30% de cadaverina (CAD) mais alta (P≤ 0,05). Aflatoxnas (AFL) (total) e zearalenone (ZEN), T-2/HT-2 tinham concentrações mais altas em respectivamente 16,00% (P≤ 0,05) e 13,40% (P≤ 0,05), 1,80% (P≤ 0,05) na silagem preparada em fazendas orgânicas. Deoxinivalenol (DON) tinha concentração mais alta 42,40% (P≤ 0,05) na silagem de fazendas convencionais. Ácidos graxos voláteis (AGV), ácido lático, etanol, pH e nitrogênio de amônia mostraram que as amostras de silagem de fazendas orgânicas e convencionais tinham boa qualidade. Nosso estudo sugere diferenças na formação biogênica de amônia ou micotoxinas em silagem de fazendas orgânicas ou convencionais mas, em geral, os valores medidos foram muito baixos para serem relevantes à saúde animal. Ademais, essas diferenças podem ser devido à diferença na matéria sólida e maturidade da planta entre as amostras de silagem orgânica e convencional.(AU)

Efectos de la temperatura, la actividad de agua y el tiempo de incubación en el crecimiento fúngico y la producción de aflatoxina B1 por aislados toxicogénicos de Aspergillus flavus en sorgo / Effects of temperature, water activity and incubation time on fungal growth and aflatoxin B1 production by toxinogenic Aspergillus flavus isolates on sorghum seeds

Sorghum, which is consumed in Tunisia as human food, suffers from severe colonization by several toxigenic fungi and contamination by mycotoxins. The Tunisian climate is characterized by high temperature and humidity that stimulates mold proliferation and mycotoxin accumulation in foodstuffs. This study investigated the effects of temperature (15, 25 and 37 °C), water activity (a w, between 0.85 and 0.99) and incubation time (7, 14, 21 and 28 d) on fungal growth and aflatoxin B1 (AFB1) production by three Aspergillus flavus isolates (8, 10 and 14) inoculated on sorghum grains. The Baranyi model was applied to identify the limits of growth and mycotoxin production. Maximum diameter growth rates were observed at 0.99 a w at 37 °C for two of the isolates. The minimum a w needed for mycelial growth was 0.91 at 25 and 37 °C. At 15 °C, only isolate 8 grew at 0.99 a w. Aflatoxin B1 accumulation could be avoided by storing sorghum at low water activity levels (≤0.91 a w). Aflatoxin production was not observed at 15 °C. This is the first work on the effects of water activity and temperature on A. flavus growth and AFB1 production by A. flavus isolates on sorghum grains.

El sorgo, que se consume en Túnez como alimento humano, puede sufrir la colonización severa de varios hongos toxicogénicos, con la consiguiente bioacumulación de micotoxinas. Además, el clima de Túnez, caracterizado por las altas temperaturas y humedad, estimula el crecimiento fúngico y la acumulación de micotoxinas en los productos alimenticios. Este estudio investigó los efectos de la temperatura (15, 25 y 37 °C), la actividad de agua (a w) (entre 0,85 y 0,99) y el tiempo de incubación (7, 14, 21 y 28 días) sobre el crecimiento y la producción de aflatoxina B1 (AFB1) de 3 aislados de Aspergillus flavus (designados como 8, 10 y 14) que se inocularon sobre granos de sorgo. El modelo Baranyi se aplicó para identificar los límites del crecimiento y la producción de micotoxinas. Las tasas máximas de crecimiento para 2 de los aislados se observaron en la combinación 0,99 a w y 37 °C. La a w mínima necesaria para el crecimiento del micelio fue de 0,91 a 25 °C y 37 °C. A 15 °C, solo el aislado 8 creció a 0,99 a w, pero fue incapaz de producir la aflatoxina B1. Es posible evitar la acumulación de aflatoxina B1 en el sorgo almacenándolo a baja actividad de agua (≤ 0,91 a w). Este es el primer trabajo que ha estudiado el efecto de la actividad del agua y la temperatura sobre el crecimiento de aislados de A. flavus y su producción de aflatoxina B1 en granos de sorgo.

Presencia de Fusarium graminearum en muestras de trigo destinado al consumo humano / Fusarium graminearum presence in wheat samples for human consumption

La fusariosis es una de las enfermedades más importantes de los cereales, Fusarium graminearum es su principal agente etiológico. Este hongo posee la capacidad de producir distintos tipos y niveles de toxinas, en especial deoxinivalenol (DON). En la campaña 2012-2013 se dieron condiciones ambientales predisponentes para el desarrollo de esta enfermedad. El objetivo de este trabajo fue evaluar la presencia del hongo y el contenido de DON en 50 muestras de trigo. Los resultados demostraron la presencia de Fusarium graminearum en el 80 % de las muestras analizadas. El 24 % de las muestras presentó valores de DON ≥ 1µg/g, el 26 % varió entre 0,5 y 0,99µg/g, mientras que el 50 % restante mostró valores inferiores a 0,5µg/g. Se observó correlación entre la presencia de Fusarium graminearum y de DON. Es necesario establecer valores límites de DON en granos de trigo destinados al consumo humano

One of the most important diseases in cereal crops is Fusarium head blight, being Fusarium graminearum the main etiological agent. This fungus has the ability to produce a wide spectrum and quantity of toxins, especially deoxynivalenol (DON). During the last crop season (2012-2013) the climatic conditions favored Fusarium colonization. The objective of this work was to determine the presence of this fungus as well as the DON content in 50 wheat grain samples. Our results showed that 80% of the samples were contaminated with Fusarium graminearum. Twenty four percent (24%) of the samples contained ≥ 1µg/g DON, 26% ranged from 0,5 and 0,99µg/g, and the remaining 50% had values lower than 0,5µg/g. Correlation was found between the presence of Fusarium graminearum and DON. It is necessary to establish DON limit values in wheat grains for human consumption

Hongos tóxicos en la Ciudad de Buenos Aires y alrededores / Toxic fungi in Buenos Aires City and surroundings

En la Facultad de Ciencias Exactas y Naturales de la Universidad de Buenos Aires (FCEN-UBA) funciona el Servicio de Identificación de Hongos Tóxicos. Lo integran los investigadores del Programa de Plantas Medicinales y Programa de Hongos que Intervienen en la Degradación Biológica (PROPLAME-PRHIDEB, CONICET) y colabora con servicios médicos, estatales y privados, identificando los materiales remitidos en casos de intoxicaciones con hongos, permitiendo, en muchos casos, realizar el tratamiento adecuado. El presente trabajo da a conocer los casos atendidos por el servicio desde 1985 hasta 2012 inclusive, además de una tabla para reconocer las especies tóxicas más comunes de la región. Según esta información, el 47% de las consultas que se recibieron correspondieron a pacientes menores de 18 años de edad que ingirieron materiales fúngicos de forma accidental (o al menos se sospechaba que lo hubieran hecho). El 53% restante correspondió a adultos que afirmaron ser capaces de distinguir hongos comestibles de tóxicos. Se determinó que Chlorophyllum molybdites fue la principal especie causante de intoxicaciones, la cual es comúnmente confundida con el hongo comestible Macrolepiota procera. En segundo lugar Amanita phalloides, un hongo altamente tóxico, que se caracteriza por presentar inicio de síntomas en forma tardía (latencia de 6-10 horas), evolucionando a falla hepática con el consiguiente requerimiento de trasplante o la muerte, si no se realiza el tratamiento adecuado en forma oportuna.

In Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires there is a service called Servicio de Identificación de Hongos Tóxicos, directed by researchers of the Program of Medicinal Plants and Fungi Involved in Biological Degradation (PROPLAME-PRHIDEB, CONICET) that assist hospitals and other health establishments, identifying the different samples of fungi and providing information about their toxicity, so that patients can receive the correct treatment. The objective of the present study was to analyze all the cases received from 1985 to 2012. This analysis permitted the confection of a table identifying the most common toxic species. The information gathered revealed that 47% of the patients were under 18 years of age and had eaten basidiomes; the remaining 53% were adults who insisted that they were able to distinguish edible from toxic mushrooms. Chlorophyllum molybdites turned out to be the main cause of fungal intoxication in Buenos Aires, which is commonly confused with Macrolepiota procera, an edible mushroom. In the second place Amanita phalloides was registered, an agaric known to cause severe symptoms after a long period of latency (6-10 hours), and which can lead to hepatic failure even requiring a transplant to prevent severe internal injuries or even death, is not early and correctly treated.

Mycotoxin in medicinal/aromatic Herbs: a review / Micotoxinas en hierbas medicinales y/o aromaticas: Revisión

Owing to their conditions of production, and sometimes to poor storage and transportation conditions, herbal products are susceptible to fungal contamination and development. This can lead to the accumulation of mycotoxins in this kind of commodities. Consequently, herbal products can be contaminated with fungal toxins, such as aflatoxins, ochratoxins and other mycotoxins, which pose a serious risk to public health. This paper reviews the main aspects regarding mycotoxin contamination of medical/aromatic herbs, in the context of the importance of this product in a global market. Moreover, the effect of processing on final contamination of derived foods, as well as the analytical methodology commonly employed in fungal and mycotoxin analysis in this type of products was reviewed.

Debido a sus condiciones habituales de producción, así como a condiciones inadecuadas de almacenamiento y de transporte, los productos a base de plantas medicinales o aromáticas son susceptibles a la contaminación por mohos, lo que puede conducir a la acumulación de toxinas fúngicas tales como las aflatoxinas, ocratoxinas y otras micotoxinas, lo que supone un grave riesgo para la salud pública. Este artículo revisa los principales aspectos de la contaminación por micotoxinas de las hierbas medicinales y/o aromáticas, teniendo en cuenta el contexto de este producto en un mercado globalizado, así como el efecto del procesado sobre la contaminación final de estos productos. También se revisa la metodología más comúnmente empleada en el análisis de mohos y micotoxinas en este tipo de productos.

Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay

Monoclonal antibody (mAb, NVRQS-DON) against deoxynivalenol (DON) was prepared. DON-Ag coated enzyme linked immunosorbent assay (ELISA) and DON-Ab coated ELISA were prepared by coating the DON-BSA and DON mAb. Quantitative DON calculation ranged from 50 to 4,000 ng/mL for DON-Ab coated ELISA and from 25 to 500 ng/mL for DON-Ag coated ELISA. 50% of inhibitory concentration values of DON, HT-2, 15-acetyl-DON, and nivalenol were 23.44, 22,545, 5,518 and 5,976 ng/mL based on the DON-Ab coated ELISA. Cross-reactivity levels of the mAb to HT-2, 15-acetyl-DON, and nivalenol were 0.1, 0.42, and 0.40%. The intra- and interassay precision coefficient variation (CV) were both <10%. In the mAb-coated ELISA, mean DON recovery rates in animal feed (0 to 1,000 microg/kg) ranged from 68.34 to 95.49% (CV; 4.10 to 13.38%). DON in a buffer solution (250, 500 and 1,000 ng/mL) was isolated using 300 microg of NVRQS-DON and 3 mg of magnetic nanoparticles (MNPs). The mean recovery rates of DON using this mAb-MNP system were 75.2, 96.9, and 88.1% in a buffer solution spiked with DON (250, 500, and 1,000 ng/mL). Conclusively we developed competitive ELISAs for detecting DON in animal feed and created a new tool for DON extraction using mAb-coupled MNPs.

Occurrence of aflatoxin B1 in natural products

The media claims for the consumption of natural resource-based food have gradually increased in both developing and developed countries. The interest in the safety of these products is partially due to the possible presence of toxigenic fungi acting as mycotoxin producers, such as aflatoxins produced during the secondary metabolism of Aspergillus flavus, A. parasiticus and A. nomius. Aflatoxins, mainly aflatoxin B1, are directly associated with liver cancer in human beings. This paper is aimed at evaluating the presence of aflatoxin B1 in a few vegetable drugs, dried plant extracts and industrialized products traded in 2010 in the city of Belo Horizonte, State of Minas Gerais, Brazil. The method used for the quantification of aflatoxin B1 was based on extraction through acetone:water (85:15), immunoaffinity column purification followed by separation and detection in high efficiency liquid chromatography. Under the conditions of analysis, the Limits of Detection and Quantification were 0.6 µg kg-1 and 1.0 µg kg-1respectively. The complete sets of analyses were carried out in duplicate. Aflatoxin B1 was noticed in a single sample (< 1.0 µg kg-1). The results revealed low aflatoxin B1contamination in the products under analysis. However, it is required to establish a broad monitoring program in order to obtain additional data and check up on the actual extension of contamination.

Penicillium species present in Uruguayan salami / Especies de Penicillium presentes en salamines uruguayos

The surface coverage of certain dry fermented sausages such as Italian salami by some species of Penicillium provides their characteristic flavor and other beneficial properties. One of them is the protective effect by means of a uniform film of white mold against undesirable microorganisms. The aim of this work was to identify and to isolate the fungal species present in mature Italian type of salami and to evaluate if it is possible to obtain some of them as starters. In addition, the effects of temperature (14 °C and 25 °C), water activity (a w) (0.90, 0.95 and 0.995) and 2.5 % sodium chloride (NaCl) on fungal growth were determined. Similarly, the proteolytic and lipolytic activity and the ability to produce toxic secondary metabolites were evaluated in order to characterize some possible starter strain. All species found belong to the genus Penicillium, including a performing starter as Penicillium nalgiovense and some potentially toxicogenic species. All the strains showed a higher growth rate at 25 °C. The production of extracellular proteases and lipases was significantly higher at 25 °C than at 14 °C with and without sodium chloride. Only Penicillium expansum produced patulin. On the other hand, Penicillium griseofulvum was the only species that produced ciclopiazonic acid but none of the strains produced penicillin. The species present on salami, Penicillium nalgiovense, Penicillium minioluteum, Penicillium brevicompactum and Penicillium puberulum were unable to produce any of the evaluated toxins. These findings suggest that some fungal isolates from the surface of salami such as P. nalgiovense are potentially useful as starters in sausage manufacture.

La cobertura de la superficie de los embutidos fermentados secos -como el salamín tipo italiano- por algunas especies de Penicillium les proporciona un sabor característico y otras propiedades beneficiosas. Una de ellas es el efecto de protección contra microorganismos indeseables, al formarse una película blanca uniforme de mohos. El objetivo de este trabajo fue aislar e identificar los hongos filamentosos encontrados en la superficie de salamines tipo italiano y evaluar la posibilidad de obtener especies para utilizarse como cultivos iniciadores. Se determinó el efecto de la temperatura, la actividad de agua y del cloruro de sodio sobre el crecimiento fúngico. La actividad proteolítica y lipolítica y la capacidad de producir metabolitos secundarios tóxicos fueron evaluadas con el fin de caracterizar algunos posibles cultivos iniciadores. Todas las cepas fúngicas aisladas e identificadas correspondieron a especies del género Penicillium, algunas benéficas, como Penicillium nalgiovense, y otras potencialmente toxicogénicas. Estas cepas tuvieron diferentes tasas de crecimiento en respuesta a las diferentes condiciones de cultivo. Todas las cepas mostraron mayor crecimiento a 25 °C. La producción de proteasas y lipasas extracelulares fue significativamente mayor a 25 °C que a 14 °C. Penicillium expansum fue la única especie que produjo patulina y Penicillium griseofulvum fue la única que produjo ácido ciclopiazónico. Ninguna de las especies produjo penicilina. Penicillium nalgiovense, Penicillium minioluteum, Penicillium brevicompactum y Penicillium puberulum no produjeron ninguna de las toxinas evaluadas. Estos resultados sugieren que algunos aislamientos fúngicos, como P. nalgiovense, son potencialmente útiles como cultivos iniciadores en la fabricación de estos productos.

Evaluación de la flora fungica del ambiente del archivo general del Estado Sucre Venezuela / Evaluation of the fungic flora of the atmosphere of the general archives of the Sucre State Venezuela

Los agentes fúngicos son capaces de descomponer materia orgánica como madera, papel, pintura, suelo, polvo, alimentos. Pueden ocasionar deterioro de material bibliográfico , texto, mobiliario, pinturas, igualmente causan enfermedades de tipo alérgico. El Archivo General del Estado Sucre es un centro de consulta e investigación donde se encuentran documentos y otros textos que se han ido dañando a través del tiempo; las condiciones de almacenamiento de un área con deficiencias estructurales y ambientales, favorecen el desarrollo de hongos, causando el deterioro dl material bibliográfico. Se realizaó una investigación para evaluar la flora fúngica tanto del mobiliario como documentos y textos existentes, que se servirá de base para un futuro plan de conservación y restauración. Utilizando el método de sedimentación en placa conteniendo Agar Sabouraud Dextrosa, se expusieron 60 placas durante 15 minutos en diferentes secciones del área de estudio, incubándose 10 días a temperatura ambiente. Se aislaron 86 colonias que se caracterizaron macro y microcóspicamente. Los géneros aislados fueron: Aspergillus (38,46%), Fusarium (21,70%), Curcularia (16,76%), Cladosporium (12,76%), Penicillium (10,32%), los cuales han sido descritos como contaminantes habituales de libros o textos, mobiliarios, pinturas, además capaces de causar reacciones alérgicas de tipo respiratorios resultando un riesgo para el material bibliográfico, usuarios y personal de la institución.

The fungies agents are able to disturb organics material as wood, paper, paint, ground, drust, foods they can produce deterioration of bibliographycal material, texts, house-hold goods, paintings, and equally they cause disease of alergic type. The general archives of Sucre state is a consulting center and investigation where it is found historic documents and another texts which have been damaging throuth the time, the conditions of storing in an area with estructural deficiencis and environment improve the development of fungies causing the depreciation of bibliographycal material. An investigation was done to evaluate the fingis flora as the house-hold godos, as the existen documents and texts, that it would serve the base to desing the future plan of conservation and restauration. Using the method of sedimentation in plaque containing Agar Sabouraud Dextrosa, 60 plaques were exposed during 15 minutes in different sections of study area, incubating 10 days at atmosphere temperatura. 86 colonies were isolated that were characterized macro and microcospyly. The aislated fungic species were: Aspegillus (35,46%), Fusarium (21,70%), Curvalaria (16,76%), Cladosporium (12,76%), Penicillium (10,32%), which have been described as usual contaminating of book o texts, house-hold godos, paintings, in addiction to this they are able to cuse alergic rections of respiratory type, resulting a risk to the bibliographycal material, usuaries and personal of the institution.

Patulin accumulation in apples during storage by Penicillium expansum and Penicillium griseofulvum strains

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.

Determinación de zearalenona por cromatografía líquida de alta resolución: normalización de un nuevo método para raciones especiales para cerdos / Zearalenone determination in high performance liquid chromatography: a new standardization method for commercial swine rations

The aim of this work was to offer a new method of high performance liquid chronomatography (HPLC) to evaluate commercial swine rations (CSR) contaminated by zearalenone (ZEA). After ZEA extraction and purification from CSR, the samples were eluted with acetonitrile, methanol and water solvent system. The results indicated that the proposed method showed to be rapid and efficient for the detection and quantification of ZEA in CSR, since its recovery was 102.62 percent, it offered excellent precision with a coefficient of variation of 0.9992. Furthermore, it is also proposed a as a biocontrol assay for micotoxigenic fungi isolated and maintained in the laboratory. The test was performed with the killer yeast Trichosporum insectorum CBS 10422 against Fusarium sp and Aspergillus flavus, which demonstrated to be effective against the latter.

El propósito de este articulo es ofrecer un nuevo método de cromatografía líquida de alta resolución (CLAR) para evaluar las raciones especiales para cerdos (REC) contaminado con zearalenona (ZEA). Después de la extracción y purificación de ZEA, las muestras se eluyeron con acetonitrilo, metanol y agua del sistema disolvente. Los resultados indican que el método propuesto demostró ser rápido y eficaz para la detección y cuantificación de ZEA en REC, ya que sus indicadores se presentan capaces de recuperación de 102,62 por ciento, además de ofrece una excelente precision, con un çõefiCiênte de variación de 0,9992. Por otra parte, también se propone una prueba de control biológico de hongos micotoxige-nic aislados y mantenidos en el laboratorio. La prueba se realizó con la levadura killer Trichosporum insectorum CBS 10422 contra Fusarium sp y Aspergillus flavus, mostrando eficaces sólo contra Aspergillus.

Ação fungitóxica do óleo essencial de Tanaecium nocturnum (Barb. Rodr.) Bur. e K. Shum sobre o Aspergillus flavus isolado da castanha-do-Brasil (Bertholletia excelsa) / Fungitoxic action of the essential oil of Tanaecium nocturnum (Barb. Rodr.) Bur. and K. Shum on Aspergillus flavus isolated from the Brazil nut (Bertholletia excelsa)

Neste trabalho, avaliou-se a capacidade fungitóxica do óleo essencial de folhas frescas de Tanaecium nocturnum sobre o Aspergillus flavus isolado da castanha-do-brasil, por meio das técnicas de contato e fumigação. Pelos resultados dos bioensaios realizados até 10 dias de incubação, verificou-se que a inibição total do crescimento micelial ocorreu quando se utilizou o óleo essencial nas concentrações de 782 ppm (técnica de contato) e 1000 ppm (técnica de fumigação). Em ambas as técnicas, o óleo essencial inibiu a esporulação a partir da concentração de 500 ppm. Observou-se que nos cinco primeiros dias de incubação não houve diferença significativa nos resultados apresentados pelas duas técnicas estudadas, havendo a partir daí uma redução da atividade do óleo essencial nas concentrações inferiores a 1000 ppm pelo teste de fumigação. A ação fungitóxica do óleo essencial sobre o microrganismo estudado pode ser atribuída à presença do benzaldeído (composto majoritário do óleo essencial estudado), em associação com outros compostos também presentes nesse óleo essencial, tais como; álcool benzílico, benzoato de benzila e mandelonitrila.

The present work sought to evaluate the fungitoxic activity of the essential oil from fresh Tanaecium nocturnum fresh leaves on Aspergillus flavus isolated from Brazil nuts, using contact and fumigation techniques. The results of bioassays performed up to 10 days of incubation demonstrated that total inhibition of mycelial growth occurred when using the essential oil at concentrations of 782 ppm (contact technique) and 1000 ppm (fumigation technique). In both techniques, the essential oil inhibited the formation of spores at the concentration of 500 ppm. No significant difference in the results presented by the two techniques was observed in the first five days of incubation. After this period, the essential oil showed a reduction in activity at concentrations lower than 1000 ppm in the fumigation test. The fungitoxic activity of the essential oil on the organism studied can be attributed to the presence of benzaldehyde (major component of the essential oil), in combination with other compounds also present in this oil, such as, benzyl alcohol, benzyl benzoate and mandelonitrila.

Parâmetros metabólicos de ovelhas submetidas a dietas contendo aflatoxina e zearalenona com adição de glucomanano modificado / Metabolic parameters of ewes receiving diets containing aflatoxin and zearalenone with addition of modified glucomannan

As micotoxicoses são grandes causadoras de perdas produtivas em ruminantes, sendo aflatoxina (AFLA) e zearalenona (ZEA) as principais micotoxinas encontradas em alimentos conservados. Estas micotoxinas apresentam efeito sobre o metabolismo animal, através da ação anabólica de metabólitos da ZEA, bem como pelas lesões hepáticas causadas pela AFLA. O objetivo deste estudo foi determinar a influência do adsorvente glucomanano modificado sobre parâmetros metabólicos de ovelhas submetidas à dietas contendo AFLA e ZEA. Foram utilizadas 34 fêmeas divididas em 6 grupos (ZEA; ZEA+ADS; AFLA; AFLA+ADS; CONTROLE+ADS; CONTROLE), recebendo 1,0 mg/kg de ZEA, 1,5 mg/kg de AFLA e/ou 2kg/ton de adsorvente. A ZEA diminuiu os níveis séricos de glicose, em relação ao CONTROLE (p<0,05), porém, o adsorvente não influenciou os níveis de glicose, não havendo diferença entre os grupos ZEA e ZEA+ADS. A ZEA aumentou os níveis de AST e GGT em relação ao grupo CONTROLE (p<0,05), sendo que os níveis de AST foram superiores no grupo ZEA (p<0,05), quando comparado ao grupo ZEA+ADS. Ainda, a aflatoxina causou uma redução nos níveis de albumina, em relação aos valores fisiológicos de ovinos. Assim, a partir destes resultados pode-se concluir que a ZEA causou alterações metabólicas em ovinos, bem como o glucomanano modificado foi eficiente em reduzir a possível agressão hepática causada por esta micotoxina, demonstrada pela diminuição nos níveis de AST.

The micotoxicoses are causing great losses of production in ruminants, being aflatoxin (AFLA) and zearalenone (ZEA) the major mycotoxins found in foods preserved. These mycotoxins have effect on the metabolism animal through the anabolic action of metabolites of the ZEA, and the liver injury caused by AFLA. The purpose of this study was to determine the influence of the sorbent modified glucomannan on metabolic parameters of sheep submitted to diets containing AFLA and ZEA. Were used 34 females divided into 6 groups (ZEA; ZEA+ADS; AFLA; AFLA+ADS; CONTROL+ADS; CONTROL), receiving 1.0 mg/kg of ZEA, 1.5 mg/kg of AFLA and/or 2kg/ton of sorbent. The ZEA decreased serum levels of glucose, for the CONTROL (p <0.05), however, the sorbent not influence the levels of glucose, with no difference between groups ZEA and ZEA+ADS. The ZEA increased levels of AST and GGT compared to group CONTROL (p <0.05), whereas the levels of AST were higher in the group ZEA (p <0.05) when compared to the group ZEA+ADS. Still, the aflatoxin caused a reduction in the levels of albumin, for physiological values of sheep. Thus, from these results it was concluded that ZEA caused metabolic alterations in sheep, and the modified glucomannan was effective in reducing the possible liver aggression caused by this mycotoxin, shown by the decrease in the levels of AST.

Mechanism of sterigmatocystin biosynthesis regulation by pH in Aspergillus nidulans

External pH constitutes one of the most important environmental factors that control growth, metabolism and differentiation in microorganisms, including fungi. We have analyzed the effect of external pH on sterigmatocystin biosynthesis in Aspergillus nidulans. It was observed in repeated experiments that alkaline pH, in opposition to acid pH, increased sterigmatocystin production and the transcript levels of aflR, the master gene that regulates expression of the sterigmatocystin cluster in A. nidulans. It is known that pH effects in fungi operate mostly through the Pal/Pac signaling pathway, originally described in Aspergillus nidulans. Accordingly, we studied the role of this signaling pathway in ST biosynthesis. It was observed that aflR transcript levels were increased in the "alkalinity mimicking" mutant pacCc14 and were minimal in the "acidity mimicking" mutant palA1. No sterigmatocystin was produced by palA1 or pacC- mutants at neither acid or alkaline pH of incubation. Finally, fluG and flbA, genes known to regulate both conidiation and sterigmatocystin synthesis upstream in the regulatory cascade, were up-regulated at alkaline pH.

Detection of fumonisin producing Fusarium verticillioides in paddy (Oryza sativa L. ) using polymerase chain reaction (PCR) / Detecção de Fusarium verticillioides produtor de fumonisina em arroz empregando reação de polimerase em cadeia

The study reports the occurrence of fumonisin producing Fusarium verticillioides in 90 samples of stored paddy (Oryza sativa L.)collected from different geographical regions ofKarnataka, India. Fumonisin producing F. verticillioides was identified based on micromorphological characteristics and PCR using two sets of primers. One set of primers was F. verticillioides species specific, which selectively amplified the intergenic space region of rDNA. The other set of primers was specific to fumonisin producing F. verticillioides. Eight paddy samples were positive for F. verticillioides. Eleven isolates obtained from these samples were capable of producing fumonisin.

O estudo relata a ocorrência de Fusarium verticillioides produtor de fumonisina em 90 amostras de arroz armazenado, coletado de diferentes regiões geográficas de Karnataka, Índia. F. verticillioides produtor de fumonisina foi identificado baseado em características micromorfológicas e PCR empregando dois sets de primers. Um dos sets era F. verticillioides especie-específico, que amplificava seletivamente a região do espaço intergênico do rDNA. O outro set de primers era especifico para F. verticillioides produtor de fumonisina. Oito amostras de arroz foram positivas para F. verticillioides. Onze isolados obtidos dessas amostras foram produtores de fumonisina.