Abstract The phenolic extracts obtained from rice bran cultivated with Rhizopus oryzae were evaluated about their abitily to inhibit the production of trichotecenes and emerging mycotoxins by Fusarium strains on sweet corn cobs (Zea mays L.). The rice bran was cultured with R. oryzae for 48 h at 30 °C. The phenolic compounds were extracted, clarified, lyophilized and ressuspend as water solution with different concentrations. The phenolic acid extracts were applied on sweet corn inoculated by F. cerealis, F. graminearum and F. poae incubated by 15 days at 25 °C and afterward the fermented corn was autoclaved and frozen. The mycotoxins were quantified by LC-MS/MS. The mycotoxin production by all Fusarium strains evaluated was reduced in relation to the control. F. poae was the most sensitive, because its production of the toxins T-2 and HT-2 and BEA was inhibited by phenolic extract 0,5% (p/v), while extract 1% (p/v) inhibited 96% of the DON production. DON, 3ADON and 15ADON production by F. graminearum and F. cerealis was also reduced. Therefore, the extract is a promising antimicotoxinogenic against Fusarium toxins production on sweet corn cobs and should be a good alternative to reuse wastes from agroindustry.

Abstract The potential use of microalgae in health has been the aim of different studies in the last years. This study investigated anti-inflammatory and antioxidant properties of three different extracts of green microalga Pediastrum boryanum in an acute inflammation model in rats. Rats were treated intraperitoneally with lyophilized biomass, the phenolic compounds and the extracellular extracts of P. boryanum before the induction of paw edema by the intraplantar injection of carrageenan. The edema and the levels of interleukin-1β and tumor necrosis factor-α were determined in the hind paw. Oxidative stress markers were analyzed in the liver and hepatic toxicity and genetic damage was evaluated in the blood. The results demonstrated that the three extracts of P. boryanum exhibited pronounced anti-oedematous property and decreased the levels of cytokines. The best results were provided by the phenolic compounds extract, that contains gallic, chlorogenic, protocathecuic and vanillic acid. A reduction in lipid peroxidation was observed after the treatment with lyophilized biomass and the extracellular extract improved the total antioxidant capacity of the liver. Moreover, no DNA damage and hepatic toxicity were observed after administration of P. boryanum extracts. In conclusion, these results suggest that P. boryanum can be an important source of anti-inflammatory compounds.

Abstract This study evaluated the role of dry fruits as a source of fungal contaminants or functional compounds. Dry apple, apricot, candied fruits, prune, and white and black raisin were evaluated regarding patulin and ergosterol content, centesimal composition, pH, acidity, pectin, and phenolic profile. The ergosterol was detected in 38% of the samples and patulin (PAT) in 89%, the apricot samples stood out most by contaminant level. Apricot, candied fruit, and prunes presented moisture values beyond the other dry fruits. The pH ranged from 3.8 to 4.6; acidity from 0.1 to 1.0%; sugar content from 1.7 to 5.5%; and levels of soluble and insoluble pectin from 1.6 to 5.1% and 10.9% to 27.3%, respectively. The apples showed a higher content of free phenolic compounds (6.6 mg of gallic acid/g) with a predominance of gallic and protocatechuic acids. Pearson's correlation showed a significant positive correlation between patulin and ergosterol content (0.99) and soluble pectin (0.78); a significant negative relation between total sugars (-0.66) and free phenolic compounds (-0.63). Fungal contamination and the occurrence of PAT show that this mycotoxin is present and, therefore, the evaluation of fungal toxins in dry fruits is recommended before claiming them to be a source of functional compounds.

Abstract In several countries halophytes are commercially cultivated in low saline or even irrigated with seawater, as well as with saline aquaculture effluent, like a sea asparagus Sarcocornia ambigua, that show a biotechnological potential for bioactive compounds production. However, their recovery from matrix is sometimes inefficient because the lignocellulosic materials difficult the solvent action when drastic conditions are not applied. The ultrasound-assisted extraction (UAE) was optimized by a central composite rotational design for recovery free phenolic compounds (FPC) from the sea asparagus S. ambigua. Optimum conditions were validated and compared with orbital shaker extraction for S. ambigua, other Brazilian halophytes (Apium graveolens, Myrsine parvifolia, Paspalum vaginatum, and Schinus terebinthifolius), soybean and rice bran. Except for P. vaginatum, soybean and rice bran, UAE yielded 18-29% higher FPC than that of the orbital shaker. Besides this analytical performance UAE method optimized is faster than the orbital shaker, providing shorter exposure of the analyst to the extractor solvent and applicable in matrices with different compositions. It was also demonstrated that halophytes species showed to be good natural sources of FPC in a better way as soybean and rice bran. This work was the first to report FPC in M. parvifolia and P. vaginatum.

ABSTRACT: Mycotoxin levels in Fusarium head blight (FHB) infections can be difficult to quantify. The relationship between mycotoxin and disease is not consistent and it is not clear if wheatpathogen interaction is of significance in regions where more than one Fusarium species with distinct trichothecene production ability co-exists. This study aimed to investigate whether a set of eight Brazilian wheat genotypes, varying in resistance according to classification by the breeder, exhibit a common or differential resistance to Fusarium graminearum (Fgra) (deoxynivalenolproducing) and Fusarium meridionale (nivalenol-producing) (Fmer) using full-spike and central spikelet inoculation (type II resistance). Fgra was generally more aggressive than Fmer based on the percentage of diseased spikes (99 and 84 %, respectively) and number of diseased spikelets (mean 2.8 and 2.0, respectively) below the central spikelet. The genotype-pathogen species interaction was not significant, but there were differences between the genotypes, with BRS 194 and BRS 327 being the least and most resistant, respectively, based on severity ratings. The incidence of Fusarium-damaged kernel (FDK, %) was not affected by species, but two genotypes (BRS Parrudo and BRS 327) showed a lower incidence of FDK. There were substantial variations in the accumulation of deoxynivalenol and nivalenol among the genotypes, reaching maxima of 691.2 µg g−1 and 355.2 µg g−1, respectively, suggesting that Fgra is a more potent producer of trichothecene. Our data confirm prior resistance classifications by the breeders and suggest that the use of a single highly aggressive Fgra isolate may be sufficient for effective screening for FHB resistance. However, further studies are needed to elucidate the accumulation of resistance to trichothecene.

Abstract The objective of this work was to determine aflatoxin M1 in fresh milk from fifty-two small farms in the city of Concórdia - SC, Brazil. Samples from the cooling tanks of each property were collected from November 2014 to January 2015. The QuEChERS method was used for the extraction of aflatoxin M1, and quantification was performed in UHPLC-FL. 40.4% of the analyzed samples (eg, 21 samples) showed contamination levels by aflatoxin M1 above the maximum limit allowed by the Brazilian regulation, which is 0.5 μg L-1. These results suggest the importance of implementing Good Practices in obtaining feed for dairy cows, since the contamination of milk by aflatoxin M1 occurs through the biotransformation of aflatoxin B1, after the ingestion of feed or silage contaminated by the animals, posing risk to the animals themselves, as well as to consumers of milk and dairy products.

This study established the conditions for the extraction of the enzyme peroxidase (PO) from soybean meal (SBM). An experimental design methodology was carried out in order to evaluate the effects of stirring rate time, pH and extracting solvent volume on the enzyme extraction. By using 5 g SBM and 50 mL phosphate buffer 10 mmol L-1 pH 4.7, 60 min stirring rate at 100 rpm, an enzyme with specific activity of 100 U mg-1 for SBM was obtained. Two techniques of purification were tested and compared for purification of peroxidase from soybean meal: triphasic partition (TPP) and molecular exclusion chromatography. TPP showcased a greater efficiency with 50% recuperation and a purification factor of 13.6. Peroxidase in crude and pure forms was characterized for kinetics, thermodynamics and biochemistry. The parameter of thermal inactivation indicates high stability to exposure time and temperature increase, showing that enzyme activity is not altered by the presence of constituents of the reaction medium. Peroxidase in crude form represented a greater upkeep in activity, keeping 50% activity for 114 days at 0 °C. Peroxidase in pure form had greater affinity for substrate and reduced Deoxynivalenol levels by 80%, 20% more than the crude form.

Embora Fumonisina B1 (FB1) seja uma micotoxina pouco estudada em trigo, os fungos do gênero Fusarium são frequentemente detectados neste cereal, principalmente em regiões subtropicais, como o Sul do Brasil, o que justifica a necessidade de avaliar sua ocorrência. Neste estudo, foram avaliadas a micobiota, atividade de água e a presença de FB1 em 11 cultivares de trigo cultivados nas regiões nordeste e noroeste do Rio Grande do Sul. A micobiota foi identificada por técnicas morfológicas e moleculares e a FB1 foi determinada por cromatografia líquida e detecção por fluorescência (HPLC-FL). Nas amostras analisadas, foi verificada uma contagem inferior a 103UFC, e os fungos dos gêneros Phoma, Cladosporium e Fusarium foram os mais frequentes. Apesar disso, 54% das amostras estavam contaminadas com FB1 em níveis variando de 958 a 4906µg de FB1 kg-1.

Although fumonisin B1 (FB1) is a mycotoxin poorly studied in wheat, the Fusarium genus fungi have been frequently detected in this cereal, especially in subtropical regions such as southern Brazil, thus studies on their occurrence are required. This study evaluated the mycobiota, water activity and the presence of FB1 in 11 wheat cultivars grown in the northeastern and northwestern of Rio Grande do Sul. The mycobiota was identified by morphological and molecular techniques and FB1 was determined by liquid chromatography with fluorescence detection (HPLC-FL). Microbial counts lower than 103CFU were found in the samples, and the fungi genera Phoma, Cladosporium and Fusarium were the most frequent. Despite this, 54% of the samples were contaminated with FB1 at levels ranging from 958 to 4,906µg FB1 kg-1.

The present study aimed to evaluate the lipid content and fatty acid profiles of 9 marine species (Nannochloropsis oculata, Thalassiosira pseudonana, Phaeodactylum tricornutum, Isochrysis galbana, Tetraselmis suecica, Tetraselmis chuii, Chaetoceros muelleri, Thalassiosira fluviatilis and Isochrysis sp.) and 1 freshwater microalga species (Chlorella vulgaris) that were cultivated in Erlenmeyer flasks with 800 mL of culture medium under stationary autotrophic conditions while exposed to a continuous photon flux density of approximately 175 umol m-2 s-1, a temperature of 25±2 ºC and constant airflow. N. oculata and T. pseudonana exhibited the highest lipid content, although they had a high proportion of monounsaturated fatty acids relative to polyunsaturated fatty acids. C. vulgaris and Isochrysis sp. exhibited high levels of polyunsaturated fatty acids, whereas P. tricornutum, N. oculata and T. suecica exhibited the highest eicosapentaenoic acid levels, and I. galbana and C. muelleri had the highest docosahexaenoic fatty acid levels. The freshwater species C. vulgaris had the highest levels of linoleic and linolenic acids, followed by the marine species T. chuii.

El presente estudio pretende como objetivo evaluar el perfil de ácidos grasos y contenido de lípidos de nueve microalgas marinas (Nannochloropsis oculata, Thalassiosira pseudonana, Phaeodactylum tricornutum, Isochrysis galbana, Tetraselmis suecica, Tetraselmis chuii, Chaetoceros muelleri, Thalassiosira fluviatilis e Isochrysis sp.) y una de agua dulce (Chlorella vulgaris) y la cultura autotrófica estacionaria, usando frascos que contienen 800 ml de medio de cultivo, proporcionando constante la densidad del flujo de fotones de iluminación en los cultivos de alrededor de 175 mol m-2 s-1, temperatura 25 ± 2 °C y aireación. La especie N. oculata y T. pseudonana mostraron los niveles más altos de lípidos, pero con una alta proporción de ácidos grasos monoinsaturados en relación con poliinsaturados. La especie C. vulgaris e Isochrysis sp. mostraron altos niveles de ácidos grasos poliinsaturados; P. tricornutum, N. oculata y T. suecica revelaron los niveles más altos de ácido eicosapentaenoico y C. muelleri e I. galbana alcanzaron los más altos niveles del ácido graso docosahexaenoico. C. vulgaris reveló un mayor contenido de ácidos linoleico y linolénico, seguido de las especies marinas T. chuii.

Four fungicide trials were conducted in northern RS state during 2009. Six treatments, from the combination of one (flowering = F) or two applications (F + 10 days) of metconazole sprayed alone or in mixture with pyraclostrobin were tested. Fusarium head blight incidence (INC) and severity (SEV) were assessed at the pre-harvest period and Fusarium-damaged kernel (FDK), kernel infection (KI) and deoxynivalenol (DON) and nivalenol (NIV) trichothecenes were quantified on harvested kernels. Disease incidence ranged from 43.8% to 90% and from 7.3% to 31% respectively in the unsprayed check. Both INC and SEV were reduced in all fungicide treatments compared to the check across trials. Levels of FDK and KI were not affected by fungicides where susceptible and moderately resistant cultivars were used, respectively. The fungicide x cultivar susceptibility interaction was significant with the highest severity reduction (>70%) susceptible cultivars. Overall yield increased (+400 kg/ha) in fungicide-treated plots compared to the check; the mixture of fungicides led to higher increase (+275kg/ha) in yield. Yield increased modestly (+150 kg/ha) for two sprays compared to a single spray. Both DON and NIV were found in variable levels across trials but non significant effect of fungicides was found on mycotoxin levels.

A aplicação de substâncias naturais com efeito antifúngico é motivada pela necessidade de alternativas aos métodos existentes que nem sempre são aplicáveis, eficientes ou sem risco de danos ao consumidor ou meio ambiente. Além disso, estudos para elucidar o comportamento de espécies toxigênicas mediante fungicidas naturais tornam-se necessárias, contribuindo dessa forma com a segurança alimentar. Neste trabalho, extrato fenólico de Spirulina foi utilizado para avaliar a atividade antifúngica sobre 12 cepas toxigênicas de Fusarium graminearum, isoladas de cevada e trigo. As rotas metabólicas que poderiam ser afetadas foram avaliadas através da determinação de compostos estruturais (glicosamina e ergosterol) e da atividade de enzimas do metabolismo primário dos micro-organismos. Os resultados indicaram que os extratos fenólicos reduziram a velocidade de crescimento das espécies toxigênicas estudadas. O IC50 foi obtido na concentração de 3% a 8% (p/p) de extrato fenólico. A utilização desse fungicida natural foi promissora para a inibição da multiplicação fúngica, em especial pela inativação dos sistemas enzimáticos (amilase e protease) de Fusarium graminearum.

The application of natural antifungal substances is motivated by the need for alternatives to existing methods that are not always applicable, efficient, or that do not pose risk to consumers or the environment. Furthermore, studies on the behaviour of toxigenic species in the presence of natural fungicides have enabled their safe application in the food chain In this study, Spirulina LEB-18 phenolic extract was assessed for its antifungal activity on 12 toxigenic strains of Fusarium graminearum isolated from barley and wheat. The susceptible metabolic pathways were assessed through the determination of structural compounds (glucosamine and ergosterol) and enzyme activity of the microorganisms' primary metabolism. The results indicate that phenolic extracts reduced the growth rate of the toxigenic species investigated. The IC50 was obtained by applying 3 to 8% (p/p) of phenolic compounds in relation to the culture medium. The use of this natural fungicide proved promising for the inhibition of fungal multiplication, especially in terms of the inactivation of enzymatic systems (amylase and protease) of Fusarium graminearum.

γ-orizanol, um fitoquímico, tem atividade antioxidante e benefícios potenciais para saúde. Esse estudo objetivou desenvolver um método para extração de alta quantidade de γ-orizanol de farelo de arroz e verificar sua estabilidade em diferentes solventes. Dentre os cinco métodos estudados, o que teve maior rendimento em γ-orizanol foi melhorado, usando um delineamento composto central rotacional (DCCR - 2(4), incluindo 8 experimentos no ponto axial e 4 no ponto central, totalizando 28 experimentos). As variáveis testadas foram temperatura, tempo, volume e proporção de hexano: isopropanol usados na extração. Elevado conteúdo de γ-orizanol (13,98 mg g-1) foi obtido quando as condições de extração foram 40° C, 40 min e 75 mL de hexano:isopropanol (1:3). Os limites de detecção e quantificação do método foram 0,9 µg g-1 e 31,0 µg g-1, respectivamente. A precisão instrumental foi 0,004%, a repetibilidade (CVr) 9,4% e a recuperação 111,7 ± 17,7%. γ-orizanol é mais estável em isopropanol do que em hexano, onde 100% da concentração foi mantida após refrigeração por 72 dias.

γ-oryzanol, a phytochemical, has antioxidant activities and potential health benefits. In this study, we aimed to develop a method for extracting high amounts of γ-oryzanol from rice bran and for verifying its stability in different solvents. Of the 5 methods under investigation, the one that yielded the highest γ-oryzanol content was further improved by using a central composite rotational design (DCCR - 2(4), including 8 tests in axial conditions and 4 central points, totaling 28 trials). The parameters under study were temperature, time, volume, and the ratio of hexane:isopropanol. High γ-oryzanol content (13.98 mg.g-1) was obtained by using the following extraction conditions: 40° C, 40 min, and 75 mL hexane:isopropanol (1:3). The limits of detection and quantification of the method were 0.9 µg g-1 and 31 µg g-1, respectively. The instrumental precision was 0.004%, the repeatability (CVr) was 9.4%, and the recovery was 111.7 ± 17.7%. γ-oryzanol is more stable in isopropanol than in hexane, in which 100% concentration could be maintained after refrigeration for 72 days.

The objective of this work was to evaluate the solid-state fermentation with Rhizopus oryzae CCT 7560 of rice bran for the enrichment of proteins and the antioxidant compounds in the fermented biomass. Fermentation was performed in tray bioreactors at 30ºC for 120 h. Protein extraction was done at alkaline pH, followed by precipitation with acetone. Phenolic compounds were extracted with cold methanol. The maximum protein was recovered from after 120 h (26.6%). The content of total phenolic compounds increased during the fermentation and was maximum after 96 h, which inhibited the DPPH radical by 87%. The promising characteristics of the protein and phenolic extracts of the biomass suggested the application in the coating composition for vegetal tissues preservation.

Este estudo comparou quatro procedimentos de extração para a determinação simultânea das micotoxinas citrinina e ocratoxina A em amostras de arroz. Os procedimentos de Soares e Rodriguez-Amaya, Tanaka e os métodos de extração QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) e por ultrassom foram comparados em termos de geração de resíduos, limite de detecção, limite de quantificação e recuperação simultânea de micotoxinas. O procedimento de Soares e Rodriguez-Amaya e a extração por ultrassom apresentaram as menores recuperações, 83 e 55% para a ocratoxina A (OTA) e 55 e 48% para citrinina (CIT), respectivamente. O procedimento de Tanaka e a extração QuEChERS tiveram recuperações de 98 e 105% para OTA e 64 e 78% para CIT, respectivamente. Este último permitiu a extração simultânea das duas micotoxinas com uma redução de até 25 vezes na quantidade de solventes.

This study compared four extraction procedures for the simultaneous determination of mycotoxins ochratoxin A and citrinin in samples of rice. Soares and Rodriguez-Amaya and Tanaka procedures and the extraction methods QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) and by ultrasound were compared in terms of residue generation, limit of detection, limit of quantification and simultaneous recovery of mycotoxins. Soares and Rodriguez-Amaya and the method by ultrasound presented the lowest recoveries, 83 and 55% for ochratoxin A (OTA) and 48 and 55% for citrinin (CIT), respectively. Tanaka procedure and QuEChERS method presented the highest recoveries, 98 and 105% for OTA and 64 and 78% for CIT, respectively. The latter enabled the simultaneous extraction of the two mycotoxins, with a reduction of up to 25 times of the amount of the involved solvent.

The present study was carried out to establish the optimal conditions for performing ochratoxin A (OTA) and citrinin (CIT) extraction using the QuEChERS method in rice. Employing the factorial experimental design, variables that significantly influenced the extraction stages were determined. The following variables were analyzed: addition of water, acidification of acetonitrile with glacial acetic acid, as well as amounts of magnesium sulfate, sodium acetate, sodium citrate and diatomaceous earth. The best combining procedure resulted in a predictive model using more water and less diatomaceous earth. Recoveries of CIT and OTA were 78-105%.