A Three-Year Study on the Nutritional Composition and Occurrence of Mycotoxins of Corn Varieties with Different Transgenic Events Focusing on Poultry Nutrition.

Corn is one of the most produced cereals in the world and plays a major role in poultry nutrition. As there is limited scientific information regarding the impact of transgenic technology on the quality and nutrient composition of the grains, this study investigated the effect of three major transgenic corn varieties-VT PRO3®, PowerCore® ULTRA, and Agrisure® Viptera 3-on the field traits, nutrient composition, and mycotoxin contamination of corn grains cultivated in southern Brazil during three consecutive harvests. VT PRO3®, while demonstrating superior crop yield, showed susceptibility to mycotoxins, particularly fumonisins. In contrast, PowerCore® ULTRA, with the lowest yield, consistently exhibited lower levels of fumonisins. VT PRO3® had higher AMEn than the other varieties, while PowerCore® ULTRA had the highest total and digestible amino acid contents over the three years. The study's comprehensive analysis reveals the distinct impact of transgenic corn technologies on both productivity and nutritional levels. Balancing the crops yield, mycotoxin resistance, and nutritional content of corn is crucial to meet the demands of the poultry feed industry. Such insights are essential for decision-making, ensuring sustainability and efficiency in agricultural production as well as meeting the demands of the poultry industry.

Adsorption of aflatoxin B1 by different antimycotoxin additives: bentonite, clinoptilolite, and beta-glucans extracted from yeast cell wall.

The present study aims to evaluate and compare antimycotoxin additives (AMAs) composed of bentonite (AMA 1), clinoptilolite (AMA 2), and beta-glucans extracted from yeast cell wall (AMA 3), with respect to their ability to bind Aflatoxin B1 (AFB1) using the isothermal models of Freundlich, Langmuir, and BET. The additives were submitted to an in vitro adsorption experiment with AFB1 (0.05-4 mg L-1), using solutions of pH 3 and pH 6, with an inclusion rate of 0.5%, and analyzed by HPLC-MS/MS. At pH 3, for the seven concentrations evaluated, AMA 1 obtained adsorption rates (99.69 to 99.98%) higher (p < 0. 05) than the other AMAs, which were from 82.97 to 88.72% (AMA 2) and from 79.43 to 89.32% (AMA 3). At pH 6, in concentrations of 1, 2, and 4 mg L-1 of AFB1, AMA 1 obtained higher (p < 0.05) adsorption results (97.86 to 99.86%) than AMA 2 (91.98 to 96.12%) and AMA 3 (87.56 to 93.50%). The Freundlich model best fitted the AMA 1 adsorption data. For the other additives, the Langmuir model obtained the best fit, demonstrating qm of 8.6 mg g-1 at pH 3 and 2.3 mg g-1 at pH 6 for AMA 2; and for AMA 3, with qm of 3.4 mg g-1 at pH 3 and 2.3 mg g-1 at pH 6. The isotherm models work as an effective tool to describe the adsorption process whereas the AMA adsorption capacity varies as a function of product composition, pH, and mycotoxin content.

Monitoring of carbon dioxide and equilibrium moisture content for early detection of physicochemical and morphological changes in soybeans stored in vertical silos.

In the context of grain storage, impurities and soybeans defects in soybeans can significantly impact the equilibrium moisture content. This, cause moisture migration and heating of the stored product, leading to increased respiratory activity. Furthermore, temperature measurements within stored grain mass do not provide sufficient information for effective grain quality monitoring, primarily due to the grains excellent thermal insulating properties. To address this issue, we propose a different approach: monitoring the equilibrium moisture content and CO2 concentration as indicators of soybean respiration within the intergranular spaces of the stored grain mass. This study propose monitoring the CO2 concentration in the intergranular air along with environmental variables for early detection of physicochemical and morphological changes in soybeans stored in vertical silos using near infrared spectroscopy, X-ray diffraction and scanning electron microscopy. Thermogravimetry and spectrometry analyses revealed that the interrelationships among variables had a direct impact on soybean quality attributes. Specifically, the presence of soybeans with 5.2 % impurities led to an increased in respiration rates, resulting in a CO2 concentration of up to 5000 ppm and the consumption of up to 3.6 % of dry matter. Consequently, there were changes in the percentage of ash, proteins, fibers, and oils compositions. These findings highlight the potential for indirect assessments, enabling the prediction of physicochemical quality and contamination of soybeans stored in vertical silos through continuous monitoring of CO2 concentration and equilibrium moisture content.

Replacing Animal Fat with Gels of Psyllium Fiber and Combined Linseed Oil-Psyllium Fiber in Salamis: Impacts on Technological, Nutritional, Oxidative, and Sensory Properties.

This study produced two gels: one solely using psyllium fiber (GP) and another combining this fiber with linseed oil (GL+P). Both gels replaced 15% and 30% of the animal fat content of salamis. The objective was to evaluate the impact of this lipid reformulation on the technological, nutritional, oxidative, and sensory properties of the salamis. The lipid reformulation did not alter the evolution of pH and lactic acid bacteria during processing. The addition of GL+P did not interfere with the product's drying process. However, replacing 30% of animal fat with the GP resulted in greater weight loss and a lower final Aw value. The lipid reformulation minimally affected the color of the salamis but significantly enhanced their nutritional profile. This improvement was marked by a decrease in fat content and an increase in protein. Specifically, in the samples with GL+P, there was a rise in linolenic acid content and a reduction in the n-6/n-3 PUFA ratio. Adding GP did not affect the salamis' oxidative stability and sensory profile. However, substituting 30% of the animal fat with GL+P increased the TBARS values, and volatile compounds derived from lipid oxidation hampered the products' sensory profiles. A reduction in these negative effects was observed when replacing 15% of the fat with GL+P, suggesting this to be the ideal dosage for balancing the nutritional benefits with maintaining the product's oxidative stability.

A two-year study on the occurrence and concentration of mycotoxins in corn varieties with different endosperm textures.

BACKGROUND: Mycotoxin monitoring in cereal grains has great importance in the food and feed industries. This study evaluated mycotoxin contamination in corns with different endosperm textures in 2 years of cultivation. Samples of dent, semi-dent, flint and semi-flint corns from field experiments were analyzed by high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). RESULTS: Occurrences of fumonisins B1 (FB1 ) and B2 (FB2 ) in 2020 were 45.72% (mean 270 µg kg-1 ) and 35.89% (94.97 µg kg-1 ), respectively, and 68.98% (446 µg kg-1 ) and 45.83% (152 µg kg-1 ) in 2021. Occurrence of aflatoxin B1 was 11.96% (0.16 µg kg-1 ) in 2020 and 11.11% (0.13 µg kg-1 ) in 2021. In 2020, deoxynivalenol (DON) and zearalenona (ZEA) presented occurrences of 1.28% and 1.70%, with means of 4.08 and 2.45 µg kg-1 , respectively. In 2021, results were 8.33% (31.00 µg kg-1 ) for DON and 8.79% (4.38 µg kg-1 ) for ZEA. Citrinin, diacetoxyscirpenol and fusarenon-X did not occur in 2020 but presented 1.66%, 0.83%, and 2.50% positive rates in 2021, respectively. In 2020, flint corn presented the lowest concentration of FB1 whereas dent corn presented the highest concentration of FB1 and FB2 (P < 0.05). In 2021, dent corn presented the highest means of FB1 , FB2 and diacetoxyscirpenol (P < 0.05). Dent and semi-dent presented the highest concentration of nivalenol (P < 0.05). CONCLUSION: The endosperm texture influenced mycotoxin contamination in corn grains, especially FB1 and FB2 , which had the highest concentration in dent corn in the 2 years of this study. © 2023 Society of Chemical Industry.

Growth performance of broiler chickens fed on feeds with varying mixing homogeneity.

In feed manufacturing, the need for an effective combination of ingredients to produce a complete feed for animals is well known. However, limited quantitative data illustrate the effect of low homogeneity on the performance of animals. This study was conducted to determine the coefficient of variation (CV) of feed mixtures produced with different mixing times and the effects of these feeds with varying homogeneity consumed by 1-40-day-old broilers on their feed intake, body weight gain, feed conversion, mortality, and body weight uniformity. The mixing times used were 30, 60, 90, and 120 s, and the CV obtained from the feeds ranged from 49.5% to 5.4%. A linear decrease was observed in the feed CV with increasing mixing time, but sometimes, a longer mixing time did not present the lowest CV. The influence of the lack of feed homogeneity on the performance of broilers was mainly observed in body weight gain and, in some cases, in feed conversion. The effects on feed intake were more consistent for broilers older than 22 days, increasing body weight gain without affecting feed conversion. Our results suggest that feeds with CVs up to 22.6% had no adverse effects on the performance of broilers older than 12 days, and the lack of uniformity in broiler body weight was not influenced by feed homogeneity.

Prediction of fumonisins B1 and B2 in corn distiller's dried grains with solubles through near-infrared reflectance spectroscopy.

BACKGROUND: Distiller's dried grains with solubles (DDGS) are coproducts of the biofuel industries that use corn as raw material. This cereal is commonly contaminated by mycotoxins, including fumonisins (FBs), which can pose a serious health threat to humans and animals. Corn DDGS are typically used as a protein-rich animal feed. As mycotoxins from the original cereal grains become concentrated in DDGS, mycotoxicological monitoring is highly required before their use as ingredient in the industry. RESULTS: This work aimed to develop a methodology for predicting fumonisins B1 (FB1 ) and B2 (FB2 ) in corn DDGS using near-infrared reflectance spectroscopy (NIRS) technology associated with chemometric methods. One hundred and ninety corn DDGS samples originating from Brazilian ethanol plants and feed mills were included in this assessment. Two datasets were created: one for calibration (132 samples) and another for external validation (58 samples). Partial least squares regression and a cross-validation approach were applied to build the models. Liquid chromatography coupled to tandem mass spectrometry was used as the reference methodology. Calibration results of correlation coefficient and residual prediction deviation for FB1 and FB2 were, respectively, 0.90 and 0.88; and 2.16 and 2.06. CONCLUSION: Values of the external validation dataset were compared and no statistical difference was found between groups, indicating a satisfactory predictive ability and confirming the potential of NIRS to predict fumonisins in corn DDGS. © 2022 Society of Chemical Industry.

Development and validation of an extraction method using liquid chromatography-tandem mass spectrometry to determine patulin in apple juice.

Patulin (PAT) is a potent mycotoxin commonly found in apples and apple-based products such as juice, thus affecting global food safety. Therefore, development of fast and simple analytical methods to effectively control its contamination is of great importance. This study developed and validated a technique for the analysis of PAT in samples of industrialized apple juice based on liquid-liquid extraction and using acetonitrile as the extraction solvent. Detection via mass spectrometry was performed after Atmospheric Pressure Chemical Ionization (APCI). Mean recoveries of 97.5, 92.49 and 96.92% were reached for 4, 8 and 20 µg/L of PAT, respectively. The analyte was monitored with an APCI source in negative ion mode to identify its fragments. The 24 analyzed samples presented PAT levels below the limit of quantification. It may be concluded that the method fulfilled all of the validation criteria, thus being appropriate for routine surveillance of PAT in apple juice.

Near-infrared spectroscopy as a tool for rapid screening of deoxynivalenol in wheat flour and its applicability in the industry.

This study aimed to evaluate the applicability and efficiency of Near-Infrared Spectroscopy (NIR) by using dispersive NIR and Fourier Transform NIR to analyse 267 samples of Brazilian wheat flour contaminated with deoxynivalenol (DON). For this, Partial Least-squares Discriminant Analysis (PLS-DA) and Principal Component Analysis-Linear Discriminant Analysis (PC-LDA) were used as discriminatory methods. Next, the samples were classified according to the maximum tolerated limits (MTL) for DON in Brazil, 750 µg kg-1, and two groups were established for the calibration set: category A (≤450 µg kg-1), non-contaminated or below the MTL; and category B (>450 µg kg-1), contaminated or above the MTL. Validation samples through PLS-DA showed correct classification rates in the range of 85-87.5% and presented a 10-15% error; for PC-LDA, the hit rate was over 85% with an error of 10-15%. The present findings demonstrate that NIR is an excellent alternative method to classify wheat flour samples according to DON content.

Impact of deoxynivalenol on intestinal explants of broiler chickens: An ex vivo model to assess antimycotoxins additives.

The impact of deoxynivalenol (DON) upon intestinal tissue of broilers was assessed by using jejunal explants in Ussing chambers and analyzing histopathological and immunohistochemical parameters; this system was also applied to evaluate the efficacy of an antimycotoxins additive (AMA). The explants were subjected to the following treatments within each experiment for 120 min: Experiment 1) T1 (control) - buffer solution, and T2 - 10 mg/L DON; and Experiment 2) T1 (control) - buffer solution, T2 - 10 mg/L DON, T3 - AMA (0.5%), and T4 - 10 mg/L DON + 0.5% AMA. In Experiment 1, DON triggered a reduction in the size of enterocytes as well as of their nuclei, an increase in cytoplasmic vacuolization and apical denudation of villi. Apoptotic cells count was also greater in DON-exposed explants. In Experiment 2, the AMA mitigated DON harmful effects; cytoplasmic vacuolization of enterocytes was reduced and the size of their nuclei was preserved. The additive also promoted a partial decrease in microvillus integrity, in size of enterocytes and in apoptotic cells count. The tested ex vivo model demonstrated the impact of DON upon the intestine as well as the efficacy of the AMA against its damaging effects.

Multivariate method for prediction of fumonisins B1 and B2 and zearalenone in Brazilian maize using Near Infrared Spectroscopy (NIR).

Fumonisins (FBs) and zearalenone (ZEN) are mycotoxins which occur naturally in grains and cereals, especially maize, causing negative effects on animals and humans. Along with the need for constant monitoring, there is a growing demand for rapid, non-destructive methods. Among these, Near Infrared Spectroscopy (NIR) has made great headway for being an easy-to-use technology. NIR was applied in the present research to quantify the contamination level of total FBs, i.e., fumonisin B1+fumonisin B2 (FB1+FB2), and ZEN in Brazilian maize. From a total of six hundred and seventy-six samples, 236 were analyzed for FBs and 440 for ZEN. Three regression models were defined: one with 18 principal components (PCs) for FB1, one with 10 PCs for FB2, and one with 7 PCs for ZEN. Partial least square regression algorithm with full cross-validation was applied as internal validation. External validation was performed with 200 unknown samples (100 for FBs and 100 for ZEN). Correlation coefficient (R), determination coefficient (R2), root mean square error of prediction (RMSEP), standard error of prediction (SEP) and residual prediction deviation (RPD) for FBs and ZEN were, respectively: 0.809 and 0.991; 0.899 and 0.984; 659 and 69.4; 682 and 69.8; and 3.33 and 2.71. No significant difference was observed between predicted values using NIR and reference values obtained by Liquid Chromatography Coupled to Tandem Mass Spectrometry (LC-MS/MS), thus indicating the suitability of NIR to rapidly analyze a large numbers of maize samples for FBs and ZEN contamination. The external validation confirmed a fair potential of the model in predicting FB1+FB2 and ZEN concentration. This is the first study providing scientific knowledge on the determination of FBs and ZEN in Brazilian maize samples using NIR, which is confirmed as a reliable alternative methodology for the analysis of such toxins.

Contaminated water confirmed as source of infection by bioassay in an outbreak of toxoplasmosis in South Brazil.

The protozoan Toxoplasma gondii is a causative agent of toxoplasmosis, an important and widespread zoonotic disease. The transmission of this disease in humans includes ingestion of sporulated oocysts present in contaminated water or food. T. gondii oocysts are widely distributed and toxoplasmosis is considered a major food- and waterborne pathogen worldwide, making drinking water containing sporulated T. gondii oocysts a major source of contamination for people. In the first half of 2018, an unprecedented outbreak of toxoplasmosis was reported in the city of Santa Maria, southern Brazil. The temporal and spatial distribution of the cases strongly suggested a waterborne infection. Thus, the aim of this study was to investigate a possible involvement of treated water as a source of the outbreak. For this, piglets received potentially contaminated water ad libitum for 21 days and the infection was monitored by serology through IFAT and investigation of T. gondii DNA in tissues by PCR amplification of a 529 bp followed by mouse bioassays. All piglets receiving test water ad libitum for 21 days as well as positive controls seroconverted to T. gondii. T. gondii DNA was detected in 62.5% of the piglets that received test water. All mice inoculated with tissues from each positive piglet were PCR-positive. These results strongly indicated the presence of viable oocysts in the test water administered to the animals during the study.

Ochratoxin A production by Aspergillus westerdijkiae in Italian-type salami.

The occurrence of ochratoxin A (OTA) in matured meat products can be attributed to mycotoxin coming from raw materials and/or molds that develop on the product surface during ripening. This work aimed to evaluate OTA production by Aspergillus westerdijkiae inoculated on the surface of sausages and its diffusion into the product throughout ripening, study the effect of relative humidity (RH) on the production of this mycotoxin, and investigate the presence of OTA in dry fermented sausages that naturally present intense contamination by A. westerdijkiae. Italian-type sausages were surface inoculated with A. westerdijkiae and two distinct experiments were performed. In the first, the sausages were matured in a chamber with decreasing RH (from 95 to 75%) for 35 days. In the other, the sausages were incubated under different RH (79, 85 or 95%) for 21 days. Samples were taken at the beginning of the experiments and every 7 days, subdividing into casing, outer border, and core for analyses. Sausage samples naturally spoiled by ochratoxigenic fungi were collected during sanitary inspection. Even in the presence of A. westerdijkiae mycelia, no OTA was detected for up to 7 days of sausage maturation. On the other hand, this study demonstrated that the growth of A. westerdijkiae on salami surface produces high amounts of OTA on the casing and allows its diffusion through the casing with contamination to the outer border of sausages. In the same way, it shows that under similar water activity values of substrate, RH influences the amount of OTA produced. Conversely, OTA was restricted to the casing in the naturally contaminated sausages.

Liver transplantation and the use of KAVA: Case report.

BACKGROUND: Self-medication and the belief that herbal products are free of health risks are common in Brazil. The kava (Piper methysticum), known for its anxiolytic action, has a widespread popular use. Hepatotoxicity of kava is reported, including cases of liver transplantation and death. The kava had its use prohibited or restricted in countries like Germany, France, among others. Toxicity may be related to overdosage; however, factors such as botanical characteristics of the plant, the harvesting, storage, and production process may be associated with the development of hepatotoxic substances, such as triggering idiosyncratic reactions. HYPOTHESIS: In this case, there is a suspicion that the toxicide is intrinsic to the drug; however, the possibility of adulterants and contaminants must be ruled out. STUDY DESIGN: This study reports the case of a patient who, after using the herbal kava for 52 days, evolved into acute liver failure and liver transplantation. METHODS: The data were collected directly with the patient and compared with their clinical records. Causality was determined through the RUCAM algorithm. In addition, a phytochemical analysis of the drug used was performed. RESULTS: According to the patient's report, there is no evidence of overdosage. Results from RUCAM algorithm infer causality between liver damage and the use of kava. The analysis chemical constituents did not find any possible contaminants and major changes in the active compounds. Seven months after transplantation, the patient is well and continues to be followed up by a medical team. CONCLUSION: Our investigation indicates that there was kava-induced hepatotoxicity at standard dosages. In Brazil, self-medication by herbal medicines is frequent and many patients and health professionals do not know the risks associated with their use. Diagnosing and notifying cases in which plants and herbal medicine induce liver damage is of paramount importance to increase the knowledge about DILI and to prevent or treat similar cases quickly.

Simultaneous determination of mycotoxins in peruvian purple maize / Determinação simultânea de micotoxinas em milho roxo peruano

ABSTRACT: Purple maize is an important foodstuff for the Peruvian people. Its unique nutritional and antioxidant characteristics makes it widely exported to other countries. However, when contaminated by fungi, it can trigger numerous health problems in the consumers. This study aimed to evaluate the presence of 27 mycotoxins in 63 samples of purple maize collected in Peru. Frequency of occurrence and mean concentration of the following mycotoxins were determined: alternariolmetileter (AME), alternariol (AOH), tentoxin, neosolaniol, nivalenol, wortmannin, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, zearalenone, aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, fumonisin B1, fumonisin B2, fumonisin B3, ochratoxin A, ochratoxin α, T-2 toxin, HT-2 toxin, fusarenon x, cyclopiazonic acid, gliotoxin, agroclavin and citreoviridin. The main mycotoxins reported in purple maize were AME and AOH, with a frequency of occurrence of 14.3 and 7.9%, and mean concentration of 23.3% and 1.8%, respectively. AME and AOH do not have guidance levels in the Brazilian legislation. Contrastingly, levels of mycotoxins which are within the standards of the country's regulations were below the limit of quantification. The present results suggested that purple maize is a raw material with a great potential for the production and industrialization of special products.

RESUMO: O milho roxo é um alimento importante para o povo peruano. Suas características nutricionais e antioxidantes únicas fazem com que seja amplamente exportado para outros países. No entanto, quando contaminado por fungos, pode desencadear inúmeros problemas de saúde nos consumidores. Este estudo teve como objetivo avaliar a presença de 27 micotoxinas em 63 amostras de milho roxo coletadas no Peru. A frequência de ocorrência e a concentração média das seguintes micotoxinas foram determinadas: alternariolmetileter (AME), alternariol (AOH), tentoxina, neosolaniol, nivalenol, wortmanina, desoxinivalenol, 3-acetil desoxinivalenol, 15-acetil desoxinivalenol, zearalenona, aflatoxina B1, aflatoxina B2 , aflatoxina G1, aflatoxina G2, fumonisina B1, fumonisina B2, fumonisina B3, ocratoxina A, ocratoxina, toxina T-2, toxina HT-2, fusarenona x, ácido ciclopiazonico, gliotoxina, agroclavina e citreoviridina. As principais micotoxinas encontradas no milho roxo foram AME e AOH, com frequência de ocorrência de 14,3% e 7,9% e concentração média de 23,3% e 1,8%, respectivamente. AME e AOH não possuem níveis de orientação na legislação brasileira. Contrastantemente, os níveis de micotoxinas que estão dentro dos padrões das regulamentações do país estavam abaixo do limite de quantificação. Os presentes resultados sugerem que o milho roxo é uma matéria-prima com grande potencial para a produção e industrialização de produtos especiais.

Mycological quality of pecan nuts from Brazil: absence of aflatoxigenic fungi and aflatoxins / Qualidade micológica de nozes pecan do Brasil: ausência de fungos aflatoxigênicos e aflatoxinas

ABSTRACT: In recent years, Brazil has encouraged the cultivation of pecans to meet both demands of domestic and international market of nuts. New genetic varieties of pecans have been selected in recent years, but available scientific information on the occurrence of fungi and aflatoxins in the international literature is out of date. Therefore, the present study aimed to quantify and identify fungal microbiota and the presence of aflatoxins in pecan nuts cultivated in southern Brazil. Fifty-two pecan nut lots (Barton variety) were obtained from producers from 19 cities of Rio Grande do Sul State and analyzed by direct plating in Agar Dichloran Glycerol 18% (DG18) and Aspergillus Flavus and Parasiticus Agar (AFPA), following incubation at 25 °C for 7 days. Aflatoxins analyses were carried out using HPLC coupled with a mass spectrometer. Results revealed at least 10 different genera of fungi. Aspergillus, Penicillium, Fusarium, and Cladosporium were predominant. Xerophilic species of Aspergillus (A. wentii, A. ruber, A. pseudoglaucus, and A. chevalieri) were commonly reported in the samples. No potential aflatoxin-producing species was isolated and no aflatoxins were detected (LOQ=1 μg/kg and LOD=0.1 μg/kg for AFB1 and AFB2, and 0.3 μg/kg for AFG1 and AFG2) in the evaluated samples. The absence of this carcinogenic group of mycotoxins is highly positive and could boost the investments in the sector, as well as stimulate the commercialization and consumption of this variety of nut.

RESUMO: Nos últimos anos, o Brasil tem incentivado o cultivo de pecãs para atender a demanda do mercado nacional e internacional de nozes. Novas variedades genéticas de pecãs foram selecionadas nos últimos anos, mas informações científicas disponíveis sobre a ocorrência de fungos e aflatoxinas na literatura internacional estão desatualizadas. Portanto, o presente estudo objetivou quantificar e identificar a microbiota fúngica e a presença de aflatoxinas em nozes cultivadas no sul do Brasil. Cinquenta e dois lotes de nozes (variedade Barton) foram obtidos de produtores de 19 municípios do Estado do Rio Grande do Sul e analisados ​​por meio de Ágar Dicloran Glicerol 18% (DG18) e Aspergillus Flavus e Parasiticus Agar (AFPA), após incubação em 25 °C durante 7 dias. Análises de aflatoxinas foram realizadas usando HPLC acoplado a um espectrômetro de massa. Os resultados revelaram pelo menos 10 gêneros diferentes de fungos. Aspergillus, Penicillium, Fusarium e Cladosporium foram predominantes. Espécies xerofílicas de Aspergillus (A. wentii, A. ruber, A. pseudoglaucus e A. chevalieri) foram comumente encontradas nas amostras. Nenhuma espécie potencial produtora de aflatoxinas foi isolada e nenhuma aflatoxina foi detectada (LOQ=1 μg/kg e LOD=0,1 μg/kg para AFB1 e AFB2; e 0,3 μg/kg para AFG1 e AFG2) nas amostras avaliadas. A ausência desse grupo carcinogênico de micotoxinas é altamente positiva e pode impulsionar os investimentos no setor, além de estimular a comercialização e o consumo dessa variedade de nozes.

Aflatoxigenic and ochratoxigenic fungi and their mycotoxins in spices marketed in Brazil.

During their processing, spices usually remain close to the ground for drying, a fact that disposes to fungal contamination, as well as moisture transferred from the tropical environment can allow their multiplication and synthesis of mycotoxins. The objective of this study was to evaluate the presence of potentially toxigenic fungi and mycotoxins in spices marketed in Brazil. The fungal contamination was evaluated by direct plating for samples of clove, black and white peppers. Spread plate was used for the samples of rosemary, cinnamon, fennel, pepperoni pepper and oregano. Analyses were performed in triplicate in DG18 media with incubation at 25°C for 7days. The isolation and identification of fungi followed specific recommendations of culture media and incubation period for each genus. The presence of mycotoxins in spices was verified by high-performance liquid chromatography (HPLC) coupled to fluorescence. The frequency of species potentially toxigenic was high in white and black peppers with presence of both aflatoxigenic and ochratoxigenic fungi. Only rosemary and fennel showed contamination with aflatoxin B1 and there was a positive correlation (ρ<0.01) between the rosemary contamination with the presence of AFB1 and A. flavus. Even in the presence of ochratoxigenic fungi, ochratoxin A was not detected in the samples. The presence of natural components with antimicrobial activity could justify the low presence of mycotoxins, even in the presence of known toxigenic fungi in the samples. Mycotoxins were not detected in spices covered by Brazilian regulation of mycotoxins. On the other hand, these contaminants were present in other spices consumed by population and not mentioned in the regulation, which could be considered a cause to concern.

Effect of Bacillus spp. on Aspergillus westerdijkiae growth, sporulation and ochratoxin A production in green-coffee medium

Abstract Aspergillus westerdijkiae is one of the most important spoilage and toxigenic fungi contaminating coffee beans and may produce ochratoxin A (OTA), a mycotoxin that characterize a health risk to the coffee consumers. Biological control strategies can be used for prevention of fungal invasion and decrease mycotoxin exposure. The aims of this study were to evaluate the in vitro effect of three Bacillus sp. biocontrol candidates on A. westerdijkiae mycelial growth, spore counts and OTA production. A green-coffee based medium was inoculated with A. westerdijkiae and Bacillus spp. (B. safensis RF69, B. amyloliquefaciens RP103 and B. subtilis RP242) and after incubation, the fungal growth, sporulation and mycotoxin production was evaluated. Mycelial growth rate was reduced in a range between 76-95% and conidial production was also significantly decreased. All isolates were capable of reducing OTA production in a range between 62-96%. The results showed that the biocontrol candidates may be an effective control method for A. westerdijkiae and OTA in coffee.

Deoxynivalenol in wheat and wheat products from a harvest affected by fusarium head blight

Abstract Fusarium head blight is an important disease occurring in wheat, caused mainly by the fungus Fusarium graminearum. In addition to direct damage to crops, reduced quality and yield losses, the infected grains can accumulate mycotoxins (toxic metabolites originating from prior fungal growth), especially deoxynivalenol (DON). Wheat crops harvested in 2014/2015 in southern Brazil were affected by high levels of Fusarium head blight. In this context, the aim of this study was evaluate the mycotoxicological quality of Brazilian wheat grains and wheat products (wheat flour and wheat bran for DON. DON contamination was evaluated in 1,504 wheat and wheat product samples produced in Brazil during 2014. It was determined by high performance liquid chromatograph fitted to a mass spectrometer (LC-MS / MS). The results showed that 1,000 (66.5%) out of the total samples tested were positive for DON. The mean level of sample contamination was 1047 µg.kg-1, but only 242 samples (16.1%) had contamination levels above the maximum permissible levels (MPL) - the maximum content allowed by current Brazilian regulation. As of 2017, MPL will be stricter. Thus, research should be conducted on DON contamination of wheat and wheat products, since wheat is a raw material widely used in the food industry, and DON can cause serious harm to public health.

Anesthetic activity of the essential oil of Ocimum americanum in Rhamdia quelen (Quoy & Gaimard, 1824) and its effects on stress parameters

The aim of this study was to evaluate the anesthetic activity of the essential oil (EO) of Ocimum americanum L. in silver catfish (Rhamdia quelen ). In the first experiment, the depressor effects and chemical composition of the leaf EO (LEO) and inflorescence EO (IEO) were compared. Juveniles (n = 10) were placed in aquaria containing different concentrations of EO (25 - 500 mg L-1) to determine the point at which anesthesia was induced and the length of the recovery period. In the following experiment, the effects of 300 and 500 mg L-1 LEO exposure on stress parameters (plasma cortisol, glucose and sodium levels) after air exposure for 1 min were assayed. Fish (n = 10 per sampling time) were sampled immediately or transferred to anesthetic-free aquaria until sampling (15, 30, 60 or 240 min). LEO was composed mainly of β-linalool and 1,8-cineole in similar proportions, whereas IEO showed β-linalool as major compound. Anesthesia was obtained in silver catfish with 200-500 mg L-1 between 4-8 min for LEO and 6-16 min for IEO. Lower EO concentrations did not reach anesthetic stage up to 30 min. LEO used as anesthetic prevented the cortisol increase and sodium loss induced by aerial exposure. Glucose levels were raised in catfish exposed to LEO compared to basal group (not air exposed) in almost all observation times. EO of O. americanum obtained from leaves was considered suitable to anesthetic procedures due to its fast induction and handling-induced stress prevention.

O objetivo deste trabalho foi avaliar a atividade anestésica do óleo essencial (EO) de Ocimum americanum L. em jundiás (Rhamdia quelen ). No primeiro experimento, os efeitos depressores e a composição química dos óleos obtidos a partir das folhas (LEO) e das inflorescências (IEO) desta espécie vegetal foram comparados. Para isto, juvenis de jundiás (n = 10) foram transferidos para aquários contendo diferentes concentrações de EO (25 - 500 mg L-1) a fim de determinar os tempos de indução aos estágios de anestesia e de recuperação. No experimento seguinte foram determinados os efeitos da exposição a 300 e 500 mg L-1 de LEO em parâmetros de estresse (cortisol, glicose e níveis de sódio plasmáticos) após os animais serem expostos durante 1 min ao ar. Os peixes (n = 10 por tempo de amostragem) foram coletados imediatamente ou transferidos para aquários livres de anestésico até amostragem (15, 30, 60 ou 240 min). LEO demonstrou ser composto majoritariamente por β-linalol e 1,8-cineol em proporções similares, enquanto que IEO apresentou β-linalol como constituinte majoritário. O estágio de anestesia foi verificado em jundiás com 200-500 mg L-1 entre 4-8 min para LEO e 6-16 min para IEO. Menores concentrações de ambos os EO não promoveram anesthesia até 30 min. O uso de LEO como anestésico preveniu o aumento de cortisol e a perda de sódio induzida pela exposição aérea. Já a glicemia encontrava-se aumentada em peixes expostos ao LEO em comparação ao grupo basal (não exposto ao ar) na maioria dos tempos de observação. Assim, EO de O. americanum obtido a partir das folhas foi considerado adequado para procedimentos anestésicos devido a sua rápida indução e capacidade de prevenir o estresse decorrente do manuseio.