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.

Rapid detection of zearalenone in Coicis Semen based on ELISA / 中国中药杂志

Zearalenone(ZEN) is a toxic metabolite produced by Fusarium culmorum, F. graminearum, F. tricinctum, and other fungi, with estrogenic characteristics. Exposure to or ingestion of ZEN during pregnancy can cause reproductive dysfunction, miscarriage, stillbirth, and malformation, and seriously endanger human life and health. The detection methods for ZEN in the Chinese Pharmacopoeia(2020 edition) are liquid chromatography(LC) and liquid chromatography-mass spectrometry(LC-MS), and it is stipulated that ZEN should not exceed 500 μg in 1 000 g of Coicis Semen. Although these detection methods by instruments can achieve the qualitative and quantitative analysis of ZEN in Coicis Semen, their high detection cost and long periods hinder the rapid screening of a large number of samples in the field. In this study, the synthesized ZEN hapten was conjugated with bovine serum albumin(BSA) and ovalbumin(OVA) to obtain the complete ZEN antigen. By virtue of antibody preparation techniques, ZEN monoclonal antibody 4F6 was prepared, which showed 177.5%, 137.1%, and 109.7% cross-reactivity with ZEN structural analogs zearalanol, zearalenone, and α-zearalenol, respectively, and no cross-reactivity with other fungal toxins such as aflatoxin. Direct competitive enzyme-linked immunosorbent assay(dcELISA) based on ZEN monoclonal antibody 4F6 was developed for the determination of ZEN in Coicis Semen with an IC_(50) of 1.3 μg·L~(-1) and a detection range of 0.22-21.92 μg·L~(-1). The recoveries were 83.91%-105.3% and the RSD was 4.4%-8.0%. The established dcELISA method was used to determine the ZEN residuals in nine batches of Coicis Semen samples, and the results were validated by LC-MS. The correlation between the two detection methods was found to be 0.993 9, indicating that the established dcELISA could be used for the rapid qualitative and quantitative detection of ZEN residuals in Coicis Semen.

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.

Molecular characterization of mycotoxin-producing Aspergillus parasiticus and sensitivity pattern to different disinfectants

Aims@#The study was aimed to isolate and characterize the mycotoxin-producing filamentous Aspergillus parasiticus from the feed samples. The sensitivity pattern of the isolates was assessed against different disinfectants.@*Methodology and results@#Fifty different feed samples were screened for A. parasiticus isolation. Isolates were subjected to macroscopic and microscopic characterization. Polymerase chain reaction was performed to confirm the isolates at the genomic level. Mycotoxin producing potential of the isolates was assessed by thin-layer chromatography (TLC). To quantify the toxins, high performance liquid (HPLC) was employed. The antifungal potential of disinfectants was determined by the well diffusion method followed by minimum inhibitory concentration (MIC) calculation. Out of twenty isolates of A. parasiticus, 11(55%) isolates were observed positive for toxin production. Three toxigenic isolates (AspP2, AspP4 and AspP8) were selected to evaluate their susceptibility against disinfectants by well diffusion method. AspP2 produced maximum (5.90 ng/mL) toxin, followed by AspP4 (3.11 ng/mL) and AspP8 (18.47 ng/mL). Terralin showed maximum fungicidal activity with 29.66 ± 8.08 mm zone of inhibition at 0.42 μg/mL MIC. Hypochlorite and Instru Star showed 99% disinfection with 30, 60 and 90 min contact time (6 mean log reduction) for all A. parasiticus isolates. Alpha Guard inhibited growth after 15 min contact time for all the isolates.@*Conclusion, significance and impact of study@#This study provides data indicating the contamination of feed samples with mycotoxin-producing A. parasiticus isolates and their sensitivity against commercially available disinfectants. Use of these disinfectants in appropriate concentrations and time could help prevent the contamination of food, feed and healthcare settings with the fungal species.

The use of histológical parameters to assess intestinal and liver health on broilers challenged isolatedly and simultaneously with cyclopiazonic acid and aflatoxin / B1 El uso de parámetros histológicos para evaluar la salud intestinal y hepática en pollos de engorde desafiados de forma aislada y simultánea con ácido ciclopiazónico y aflatoxina B1

Abstract Mycotoxins contaminate agricultural commodities, which contaminates animals. These toxins can damage vital organs, such as the liver, as well as the epithelial tissue. Among these mycotoxins are aflatoxin B1 (AFB1) and cyclopiazonic acid (CPA), which can occur simultaneously in food. In broilers, mycotoxicosis has an economic impact due to several factors, such as low feed conversion rate, incidence of other diseases, and interference with reproductive capacity, all of which may lead to a public health problem. The aim of the present study was to histologically assess, through the I See Inside (ISI) method, harmful effects on broiler liver, duodenum, jejunum, and ileum in the presence of AFB1 and CPA isolatedly and simultaneously. Groups challenged with mycotoxins showed significant damage to both gut and liver fragments. All challenged-groups in all fragments impaired the parameters analyzed for intestinal epithelium. In the liver, AFB1 was predominantly harmful when the parameters were analyzed separately, but when analyzing the total ISI score, CPA was also found to be harmful to this organ. The other point analyzed was the great variation between the weights of the birds contaminated by mycotoxin while the negative control group presents a lesser variation.

Resumen Las micotoxinas contaminan los productos agrícolas, que a su vez contaminan a los animales. Estas toxinas pueden dañar órganos vitales, como el hígado y el tejido epitelial. Entre estas micotoxinas se encuentran la aflatoxina B1 (AFB1) y el ácido ciclopiazónico (CPA), que pueden hallarse simultáneamente en los alimentos. En los pollos de engorde, la micotoxicosis tiene un impacto económico debido a varios factores, como la baja tasa de conversión alimenticia, la incidencia de otras enfermedades y la interferencia de la capacidad reproductiva, que pueden llevar a un problema de salud pública. El objetivo de la presente investigación es la de evaluar histológicamente, a través del método "I See Inside" (ISI), los efectos nocivos sobre el hígado, duodeno, yeyuno e íleon de pollos de engorde en presencia de AFB1 y CPA de forma aislada y simultánea. Los grupos desafiados con micotoxinas presentaron un daño significativo tanto en el intestino como en los fragmentos del hígado. Todos los grupos tratados tuvieron alteraciones en los parámetros analizados para el epitelio intestinal. En el hígado, AFB1 fue predominantemente dañino cuando los parámetros se analizaron por separado, pero al examinar la puntuación ISI total, también se encontró que el CPA era perjudicial para este órgano. Otra cuestión que fue investigada fue la gran variación entre los pesos de las aves contaminadas por micotoxinas mientras el grupo de control negativo presentó una variación menor.

Micotoxinas en el análisis de los alimentos: Un estudio con indicadores bibliométricos y mapas de visualización / Mycotoxins in food analysis: A study with bibliometric indicators and visualization maps

El presente estudio tuvo como objetivo identificar el desarrollo de la producción científica según año y revistas; determinar la estructura temática e identificar la red de coautoría de la literatura sobre micotoxinas en el análisis de los alimentos. Se realizó un estudio descriptivo con indicadores bibliométricos y mapas de visualización, en el que se usó la base de datos Scopus de los documentos incluidos desde el año 2011 al 2020. Se consideraron 453 artículos que cumplieron con los criterios de inclusión, 382 artículos de investigación y 71 revisiones. El promedio de citas por artículo varió anualmente entre 6 (2019) y 40,5 (2012). La revista más productiva fue Mycotoxin Research, de la editorial Springer Nature. En el mapa de coocurrencias se obtuvieron 5 clústeres de descriptores. En el mapa de coautoría se hallaron 21 clústeres, 14 de ellos correspondieron a autores que investigaron en coautoría. El desarrollo científico tuvo una producción anual no uniforme, con crecimiento lento. La estructura temática según la red de coocurrencia indica que el término más frecuente es micotoxinas, correspondiente al enfoque micotoxinas que influyen en la salud pública del primer clúster. La red de coautoría indica que el 98,2 por ciento de los artículos fueron elaborados en coautoría y el autor más productivo fue Krska Rudolf, de la Queen's University Belfast, con 24 artículos(AU)

The purpose of the study was to identify the development of scientific production by year and journal, determine the thematic structure and identify the co-authorship network of the literature about mycotoxins in food analysis. A descriptive study was conducted of documents published in the database Scopus from 2011 to 2020, using bibliometric indicators and visualization maps. A total 453 articles were included which met the inclusion criteria. Of these, 382 were research articles and 71 were reviews. The average annual number of citations per article ranged from 6 in 2019 to 40.5 in 2012. The most productive journal was Mycotoxin Research from Springer Nature publishers. The co-occurrence map displayed five descriptor clusters, whereas the co-authorship map displayed 21 clusters, 14 of which corresponded to authors researching in co-authorship. Scientific development had a non-uniform, slowly growing annual output. The thematic structure according to the co-occurrence network shows that the most frequent term is mycotoxins, corresponding to mycotoxins influencing public health from the first cluster. The co-authorship network shows that 98.2percent of the articles were co-authored, and the most productive author was Krska Rudolf, from Queen's University, Belfast, with 24 articles(AU)

Identification and Production of Beauvericin by Fusarium subglutinansand F. sacchari from Sugarcane

Abstract Fusarium is producing several important mycotoxins including beauvericin (BEA). Two species of Fusarium viz. F. subglutinans and F. sacchari cause the Pokkahboeng disease of sugarcane. The studies on the occurrence and toxicity of BEA are scarce. Therefore, this study aimed to identify the isolates of Fusarium and detect their ability to produce BEA. The toxicity of BEA was also tested on brine shrimp Artemia salina. Many isolates of Fusarium were isolated from the infected plants of sugarcane in Malaysia. We identified the species of Fusarium according to their morphological characteristics. The capability of Fusarium isolates for producing the BEA was estimated by using a thin layer chromatography. The toxicity bioassay of BEA was conducted on the brine shrimp larvae. The results were identified on F. subglutinans and F. sacchari in 55 isolates of Fusarium. All isolates demonstrated the ability to produce BEA. Interestingly, BEA exhibited variation in toxicity between low toxic to very higher toxicity 100%. F. subglutinans and F. sacchari were able to produce BEA and possibly BEA may be causing toxicity in the host tissue and may be acting as a potential pathogenicity factor. Therefore, we consider BEA as an interesting factor in determining the virulence of fusarium isolate.

Clinical findings of equine leukoencephalomalacia / Achados clínicos de leucoencefalomalácia equina

Equine leukoencephalomalacia (LEM) is a disease caused by the ingestion of food, especially corn, contaminated by fumonisin, a Fusarium verticillioides (synonymous with F. moniliforme) metabolite. The clinical signs of brain injuries have an acute onset and rapid evolution. This study aimed to describe the clinical findings in 11 animals diagnosed with LEM, including cerebrospinal fluid (CSF) analysis. Of these animals, 91% (10/11) were horses, and only 9% (1/11) were asinine. The clinical localization of the lesions was 64% (7/10) cerebral, manifested mainly by altered mental state and behavioral disturbance, and 36% (4/11) were brainstem lesions, manifested by incoordination, head tilt, nystagmus, facial hypoalgesia, difficulty in apprehension, chewing, and swallowing food. Postmortem findings revealed that 82% (9/11) of the lesions were in the cerebrum and 18% (2/11) in the brainstem. CSF findings, such as xanthochromia (43%, 3/7), hyperproteinorrachia (50%, 3/6), and pleocytosis (43%, 3/7) were observed. The affected animals showed neurological signs that were compatible with cerebral and/or brainstem injuries. The CSF from animals with LEM may present with xanthochromia, hyperproteinorrachia, and pleocytosis, reinforcing the fact that this disease should be included in the differential diagnosis of encephalomyelopathies.(AU)

A leucoencefalomalácia (LEM) é uma enfermidade que acomete equídeos causada pela ingestão de milho e seus derivados e feno contaminados pela micotoxina fumonisina, um metabólito do fungo Fusarium verticillioides (sinônimo para F. moniliforme). Os sinais clínicos apresentam início agudo e evolução rápida e são decorrentes de lesões encefálicas. O objetivo deste estudo é descrever os achados clínicos de 11 equídeos diagnosticados com LEM, incluindo a análise do líquido cefalorraquidiano (LCR). 91% dos animais afetados eram equinos e somente 9% (1/11) era asinino. A localização clínica das lesões era 64% (7/10) cerebrais, manifestadas por alterações no estado mental e comportamento e 36% (4/10) no tronco encefálico, manifestadas por incoordenação, desvio lateral de cabeça, nistagmo, hipoalgesia da face e dificuldade de apreensão, mastigação e deglutição de alimentos. Comparativamente, os achados post mortem revelaram que 82% (9/11) das lesões eram no cérebro e 18% (2/11) no tronco encefálico. Alterações no LCR, tais como xantocromia (43%, 3/7), hiperproteinorraquia (50%, 3/6) e pleocitose (43%, 3/7), foram observadas. Os animais afetados apresentaram sinais clínicos compatíveis com lesões encefálicas e/ou de tronco cerebral. O LCR de animais com LEM pode apresentar xantocromia, hiperproteinorraquia, e pleocitose, reforçando que esta doença deve ser incluída como diagnóstico diferencial de encefalomielites.(AU)

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.

Isolation of Fusarium and identification of its toxins from tuberous root of Pseudostellaria heterophylla / 中国中药杂志

Fusarium is the major pathogen of root rot of Pseudostellaria heterophylla. This study aims to explain the possible distribution of Fusarium species and the contamination of its toxin-chemotypes in tuberous root of P. heterophylla. A total of 89 strains of fungi were isolated from the tuberous root of P. heterophylla. Among them, 29 strains were identified as Fusarium by ITS2 sequence, accounting for 32.5%. They were identified as five species of F. avenaceum, F. tricinctum, F. fujikuroi, F. oxysporum, and F. graminearum based on β-Tubulin and EF-1α genes. LC-MS/MS detected 18, 1, and 5 strains able to produce ZEN, DON, and T2, which accounted for 62.1%, 3.4%, and 17.2%, respectively. Strain JK3-3 can produce ZEN, DON, and T2, while strains BH1-4-1, BH6-5, and BH16-2 can produce ZEN and T2. PCR detected six key synthase genes of Tri1, Tri7, Tri8, Tri13, PKS14, and PKS13 in strain JK3-3, which synthesized three toxins of ZEN, DON, and T2. Four key synthase genes of Tri8, Tri13, PKS14, and PKS13 were detected in strains BH1-4-1, BH6-5, and BH16-2, which were responsible for the synthesis of ZEN and T2. The results showed that the key genes of toxin biosynthesis were highly correlated with the toxins produced by Fusarium, and the biosynthesis of toxin was strictly controlled by the genetic information of the strain. This study provides a data basis for the targeted prevention and control of exo-genous mycotoxins in P. heterophylla and a possibility for the development of PCR for rapid detection of toxin contamination.

Screening of zearalenone-degrading bacteria and analysis of degradation conditions / 中国中药杂志

Zearalenone(ZEN) is a mycotoxin produced by Fusarium, possessing estrogen-like effects, carcinogenicity, and multiple toxicities. To seek more efficient and practical agents for biological detoxification and broaden their application, this study isolated 194 bacterial strains from the moldy tuberous root of Pseudostellaria heterophylla, which were co-cultured with ZEN. An efficient ZEN-degrading strain H4-3-C1 was screened out by HPLC and identified as Acinetobacter calcoaceticus by morphological observation and molecular identification. The effects of culture medium, inoculation dose, culture time, pH, and temperature on the degradation of ZEN by H4-3-C1 strain were investigated. The mechanism of ZEN degradation and the degrading effect in Coicis Semen were discussed. The degradation rate of 5 μg·mL~(-1) ZEN by H4-3-C1 strain was 85.77% in the LB medium(pH 6) at 28 ℃/180 r·min~(-1) for 24 h with the inoculation dose of 1%. The degradation rate of ZEN in the supernatant of strain culture was higher than that in the intracellular fluid and thalli. The strain was inferred to secret extracellular enzymes to degrade ZEN. In addition, the H4-3-C1 strain could also degrade ZEN in Coicis Semen. If the initial content of ZEN in Coicis Semen was reduced from 90 μg·g~(-1) to 40.68 μg·g~(-1), the degradation rate could reach 54.80%. This study is expected to provide a new strain and application technology for the biological detoxification of ZEN in food processing products and Chinese medicinal materials.

Detección de aflatoxina M1 en muestras de leche cruda de vacas en tanques de enfriamiento en Boyacá (Colombia) / Detection of aflatoxin M1 in raw cow milk samples in cooling tanks in Boyacá (Colombia)

RESUMEN Una de las principales formas de contaminación de la leche con micotoxinas es el consumo de alimentos fermentados que se encuentran contaminados con mohos principalmente de Aspergillus spp., los cuales producen toxinas que pueden llegar a constituirse como un problema para la salud publica debido a su estabilidad térmica y química. El objetivo del presente trabajo fue detectar las concentraciones de aflatoxina M1 en muestras de leche de vacas en tanques de enfriamiento en cuatro municipios del departamento de Boyacá durante un año, determinando las variaciones de acuerdo con la temporada. Se realizó un estudio de corte longitudinal, descriptivo cuantitativo. Se seleccionaron aleatoriamente cuatro tanques de enfriamiento de cuatro municipios distintos del departamento; cada uno se muestreó dos veces al mes durante todo el período de estudio y se procesaron mediante metodología Charm Ez Lite . Se realizó un ANDEVA para determinar las diferencias estadísticas entre las concentraciones de la aflatoxina M1 por cada trimestre. Se determinaron diferencias estadísticas entre cada uno de los trimestres del estudio encontrando un porcentaje de positividad de 74,06% del total de muestras positivas en los trimestres de verano. 28,12% (108) de las muestras tomadas durante todo el estudio fueron positivas, con concentraciones de la toxina que oscilaron entre 0,5 y 2,0 μg/Kg de leche. Se determinó por primera vez en el departamento de Boyacá las concentraciones y variaciones estacionales de aflatoxina M1 en muestras de tanques de enfriamiento de leche, encontrando las mayores concentraciones y número de casos positivos de aflatoxina M1 en los meses de verano.

ABSTRACT One of the main forms of contamination of milk with mycotoxins is the consumption of fermented foods that are contaminated with mold, mainly Aspergillus spp, which produce toxins that can become a public health problem due to their thermal and chemical stability. The objective of the present work was to detect aflatoxin M1 concentrations in cows' milk samples in cooling tanks in four municipalities of the department of Boyacá for one year, determining the variations according to the season. A longitudinal, quantitative descriptive study was carried out, four cooling tanks from four different municipalities in the department were randomly selected, each tank, in each municipality, was sampled twice a month throughout the study period and processed using Charm methodology Ez Lite®, an ANDEVA was performed to determine the statistical differences between aflatoxin M1 concentrations for each quarter. Statistical differences were determined between each of the quarters of the study, finding a positivity percentage of 74.06% of the total positive samples in the summer quarters. 28.12% (108) of the samples taken throughout the study were positive, with toxin concentrations ranging between 0.5 and 2.0 μg/Kg of milk. Seasonal concentrations and variations of aflatoxin M1 in milk cooling tank samples were determined for the first time in the department of Boyacá, finding the highest concentrations and number of positive cases of aflatoxin M1 in the summer months.

Toxigenic fungal species and natural occurrence of mycotoxins in crops harvested in Argentina / Especies fúngicas toxigénicas y ocurrencia natural de micotoxinas en cultivos y productos cosechados en Argentina

Abstract Mycotoxins are secondary metabolites produced by fungal species that mainly belong to Aspergillus, Fusarium, Penicillium and Alternaria, which can grow in a variety of crops including cereals, oilseeds and fruits. Consequently, their prevalence in foods and by-products not only affects human and animal health but also causes important losses in both domestic and international markets. This review provides data about toxigenic fungal species and mycotoxin occurrence in different crops commonly grown in Argentina. This information will be relevant to establish adequate management strategies to reduce the impact of mycotoxins on human food and animal feed chains and to implement future legislation on the maximum permitted levels of these fungal metabolites.

Resumen Las micotoxinas son metabolitos secundarios producidos por diferentes especies fúngicas pertenecientes, principalmente, a los géneros Aspergillus, Fusarium, Penicillium y Alternaria. Dichos microorganismos pueden crecer en una gran variedad de cultivos, entre los que se incluyen cereales, oleaginosas y frutas. La presencia de micotoxinas en alimentos y subproductos no sólo afecta la salud humana y animal, sino que también causa pérdidas importantes en los mercados nacionales e internacionales. Esta revisión proporciona datos sobre la prevalencia de especies fúngicas toxigénicas y de micotoxinas en diferentes cultivos y productos cosechados en Argentina. Dicha información será relevante para establecer estrategias de manejo adecuadas para reducir la entrada de las micotoxinas en las cadenas alimentarias del hombre y de los animales, así como para establecer futuras legislaciones sobre los niveles máximos permitidos de dichos metabolitos.

Incidência de aflatoxinas B1, B2, G1 e G2 em amendoins / Incidence of aflatoxins B1, B2, G1 and G2 in peanuts

Micotoxinas são substâncias tóxicas produzidas por fungos e encontradas nos alimentos. As micotoxinas mais tóxicas são as aflatoxinas, produzidas, principalmente por Aspergillus flavus. Estudos realizados no país demonstraram alta incidência dessas micotoxinas em produtos de amendoim, que representa risco à saúde da população. O objetivo do estudo foi avaliar a incidência de aflatoxinas B1, B2, G1 e G2 em amostras de amendoins comercializados na região Nordeste do Estado de São Paulo nos períodos de 1994-2001 e 2016-2017. O método utilizado para analisar as amostras no primeiro período foi extração líquido-líquido e cromatografia em camada delgada e no segundo foi utilizando colunas de imunoafinidade, cromatografia líquida com derivatização pós- coluna e detector por fluorescência. No levantamento de 1994-2001 das 82 amostras, 39% tiveram contaminação de aflatoxinas variando de 11 a 1556 µg/kg com 37% das amostras contendo níveis maiores que 20 µg/kg, enquanto na pesquisa de 2016-17, das 56 amostras, 38% apresentaram contaminação destas toxinas variando de 0,09 a 60,40 µg/kg com 13% das amostras contendo níveis maiores que 20 µg/kg. Os resultados dos dois períodos estudados indicam que houve uma diminuição na incidência e nível das aflatoxinas estudadas, embora esta contaminação em amendoim permaneça um problema de saúde pública. (AU)

Mycotoxins are toxic compounds produced by fungi found in food. The most toxic mycotoxins are the aflatoxins produced mainly by Aspergillus flavus . Studies carried out in Brazil showed a high incidence of these mycotoxins in peanut products, a fact that represents public health problems. The aim of the study was to evaluate aflatoxins B1, B2, G1, and G2 in samples of peanuts sold in cities of the Northeast of the State of São Paulo in the period from 1994 to 2001 and from 2016 to 2017. The samples of the first period were analyzed using liquid-liquid extraction and thin-layer chromatography and the second using immunoaffinity columns, post-column derivative liquid chromatography and fluorescence detector. In the 1994-2001 survey, among 82 samples, 39% presented aflatoxins contamination ranging from 11 to 1556 µg/kg with 37% with levels greater than 20 µg/kg whereas, in the 2016-17 survey, 38% of the 56 samples presented contamination of aflatoxins ranging from 0.09 to 60.40 µg/kg and 7 samples 13% containing aflatoxins levels higher than 20 µg/kg. The results indicated there was a decrease in the incidence and level of aflatoxins, but the contamination of aflatoxins in peanuts remains a public health problem. (AU)

Incidência de aflatoxinas B1, B2, G1 e G2 em amendoins / Incidence of aflatoxins B1, B2, G1 and G2 in peanuts

Micotoxinas são substâncias tóxicas produzidas por fungos e encontradas nos alimentos. As micotoxinas mais tóxicas são as aflatoxinas, produzidas, principalmente por Aspergillus flavus. Estudos realizados no país demonstraram alta incidência dessas micotoxinas em produtos de amendoim, que representa risco à saúde da população. O objetivo do estudo foi avaliar a incidência de aflatoxinas B1, B2, G1 e G2 em amostras de amendoins comercializados na região Nordeste do Estado de São Paulo nos períodos de 1994-2001 e 2016-2017. O método utilizado para analisar as amostras no primeiro período foi extração líquido-líquido e cromatografia em camada delgada e no segundo foi utilizando colunas de imunoafinidade, cromatografia líquida com derivatização pós-coluna e detector por fluorescência. No levantamento de 1994-2001 das 82 amostras, 39% tiveram contaminação de aflatoxinas variando de 11 a 1556 μg/kg com 37% das amostras contendo níveis maiores que 20 μg/kg, enquanto na pesquisa de 2016-17, das 56 amostras, 38% apresentaram contaminação destas toxinas variando de 0,09 a 60,40 μg/kg com 13% das amostras contendo níveis maiores que 20 μg/kg. Os resultados dos dois períodos estudados indicam que houve uma diminuição na incidência e nível das aflatoxinas estudadas, embora esta contaminação em amendoim permaneça um problema de saúde pública.

Mycotoxins are toxic compounds produced by fungi found in food. The most toxic mycotoxins are the aflatoxins produced mainly by Aspergillus flavus. Studies carried out in Brazil showed a high incidence of these mycotoxins in peanut products, a fact that represents public health problems. The aim of the study was to evaluate aflatoxins B1, B2, G1, and G2 in samples of peanuts sold in cities of the Northeast of the State of São Paulo in the period from 1994 to 2001 and from 2016 to 2017. The samples of the first period were analyzed using liquid-liquid extraction and thin-layer chromatography and the second using immunoaffinity columns, post-column derivative liquid chromatography and fluorescence detector. In the 1994-2001 survey, among 82 samples, 39% presented aflatoxins contamination ranging from 11 to 1556 μg/kg with 37% with levels greater than 20 μg/kg whereas, in the 2016-17 survey, 38% of the 56 samples presented contamination of aflatoxins ranging from 0.09 to 60.40 μg/kg and 7 samples 13% containing aflatoxins levels higher than 20 μg/kg. The results indicated there was a decrease in the incidence and level of aflatoxins, but the contamination of aflatoxins in peanuts remains a public health problem.

Micotoxinas e seus efeitos à saúde humana: revisão de literatura / Mycotoxins and their effects on human health: literature review

Fungos são microrganismos que se desenvolvem em ambientes úmidos, produzindo, como metabolito secundário, micotoxinas, que contaminam alimentos e trazem danos à saúde. As micotoxinas são metabólicos produzidos por determinados gêneros de fungos filamentosos que acometem os produtos alimentícios e que, quando ingeridos, podem causar distúrbios ao organismo humano. Fungos se desenvolvem nos alimentos em qualquer fase do processo tendo condições favoráveis a eles, e uma vez que o alimento seja acometido pelo fungo as micotoxinas podem permanecer no alimento após a morte do fungo que as produz. Logo se faz pensar nos produtos ofertados. Este estudo constitui-se de uma revisão da literatura especializada, enfatizando os danos que os alimentos com micotoxinas podem causar ao organismo do ser humano. Para a coleta utilizaram-se o banco de dados Medline e os sites National Libray of Medicine (PubMed) e Scientific Eletronic Libray Online (SciELO), por meio das palavras-chave: fungos filamentosos; micotoxinas; produtos alimentícios. Sabe se que há um vasto número de micotoxinas, mas visando não só a perda econômica, mas também a saúde, pesquisas vêm se concentrando em algumas micotoxinas por apresentarem danos significativos já comprovados à saúde humana e animal, sem se descartar a possibilidade de outras apresentarem risco a saúde humana.

Fungi are microorganisms that grow in humid environments, producing, as a secondary metabolite, mycotoxins that contaminate food and bring harm to health. Metabolic mycotoxins are produced by certain genera of filamentous fungi that affect food products and which when ingested can cause disturbances to the human organism. Fungi develop in foods at any stage of the process having favorable conditions for them, and once the food is affected by the fungus the mycotoxins may remain in the food after the death of the fungus that produces them. One soon thinks about the products offered. This study is a review of the specialized literature, emphasizing the damage that foods with mycotoxins can cause to the human body. For collection it was used the Medline database and the National Libray of Medicine (Pubmed) and Scientific Electronic Libray Online (SciELO) websites, through the keywords: filamentous fungi; mycotoxins; food products. It is known that there is a large number of mycotoxins, but aiming not only economic loss, but also health, research has focused on some mycotoxins due to already proven significant damage to human and animal health, without ruling out the possibility of others presenting risk to human health.

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)

The sanitary quality of Tifton 85 bermudagrass and Wrangler grass hay stored under different environmental conditions / Variações da temperatura e qualidade sanitária de f eno de capim tifton 85 e vaquero grass sob diferentes condições de armazenament

This study evaluated the variations in temperature and dry matter content in Tifton 85 bermudagrass and Wrangler grass hay stored under different environmental conditions. The fungal and mycotoxin populations in the hay production area were also evaluated. Fifty samples (30 hay bales, ten samples of organic matter [OM], and ten samples of grass [G] before baling) were used in a completely randomized design and factorial scheme with ten treatments (a combination of two classes of G: Tifton 85 bermudagrass and Wrangler grass) and three storage systems: a covered shed without side walls, covered with a clear tarp; an open environment covered with a double-sided tarp; and a ventilated closed shed. Four additional treatments, five replications, and one sample per experimental unit completed the study design. The environment influenced the temperature of the Tifton 85 bermudagrass and Wrangler grass bales, and the dry matter content of the bales during the evaluated period was above the recommended limit (800 g kg-1). The genus Penicillium occurred with the highest frequency in the Wrangler grass stored under a double-sided tarp (6.13 log CFU g-1). In the additional treatment, the OM deposited in the soil of the Wrangler grass production area exhibited a greater occurrence of the fungus Fusarium (4.66 log CFU g-1) and of the mycotoxin zearalenone (ZEA; 594.2 µg kg-1). The hay stored in the shed with a clear tarp exhibited more aflatoxin (AFLA) and deoxynivalenol (DON). The concentration of mycotoxins varied across the storage environments and between the stored species; however, the shed storage system used in hay production might also be a source of hay contamination via the incorporation of decomposing OM with mycotoxins in the hay.

Este estudo teve como objetivo avaliar as variações na temperatura e no teor de matéria seca do feno de capim Tifton 85 e Vaquero armazenados em diferentes condições ambientais. As populações de fungos e micotoxinas na área de produção de feno também foram avaliadas. Cinquenta amostras (30 fardos de feno, dez amostras de matéria orgânica [OM] e dez amostras de gramíneas [G] antes do enfardamento) foram utilizadas em delineamento inteiramente casualizado e esquema fatorial com dez tratamentos (uma combinação de duas classes de G: Tifton 85 e capim Vaquero) e três sistemas de armazenamento: galpão coberto sem paredes laterais, feno coberto com uma lona transparente; ambiente aberto e o feno coberto com uma lona dupla face; e galpão fechado ventilado. Quatro tratamentos adicionais, cinco repetições e uma amostra por unidade experimental completaram o desenho experimental. O ambiente influenciou a temperatura dos fardos de Tifton 85 e de capim vaquero, e o teor de matéria seca dos fardos durante o período avaliado foi acima do limite recomendado (800 g kg-1). O gênero Penicillium ocorreu com maior freqüência no capim Vaquero armazenado sob uma lona de dupla face (6,13 log CFU g-1). No tratamento adicional, a MO depositada no soloda área de produção de capim Vaquero apresentou maior ocorrência do fungo Fusarium (4,66 log CFU g-1) e da micotoxina zearalenona (ZEA; 594,2 µg kg-1). O feno armazenado no galpão e coberto com lona transparente apresentou maiores teores de aflatoxina (AFLA) e desoxinivalenol (DON). A concentração de micotoxinas variou entre os ambientes de armazenamento e entre as espécies armazenadas; no entanto, a contaminação do feno pode se dar pela incorporação de MO com micotoxinas presentes no campo.