Abstract The aim of this study was to evaluate the capacity of two strains of lactic acid bacteria (LAB), Lactobacillus rhamnosus and Lactococcus lactis, and a yeast strain, Saccharomyces cerevisiae, inactivated by heat (121 °C, 10 min), from binding to aflatoxin M1 (AFM1), as well as the interaction between these microorganisms, aflatoxin M1 and the Minas Frescal cheese matrix after 2 and 30 days of storage. The ability of LABs and S. cerevisiae to bind AFM1 to Minas Frescal cheese was evaluated by high performance liquid chromatography (HPLC) composed of a fluorescence detector. The interaction between these microorganisms and AFM1 was evaluated using a scanning electron microscope composed of a backscattered electron detector with a voltage of 15 kV and magnifications of 1000 ×, 5000 × and 8000 ×. The use of microorganisms as a biological method is efficient in reducing AFM1 in Minas Frescal cheese and does not affect the microbiological parameters. AFM1 reduction varied according to the microorganism used in the treatments. S cerevisiae showed greater capacity to bind AFM1 over time, compared to LABs. Scanning electron microscopy was especially useful, confirming that lactic acid bacteria and S. cerevisae were able to bind AFM1 particles in Minas Frescal cheese.

Abstract Mycotoxins are products of the secondary metabolism of fungi, which can be present in food as contaminants. According to Food and Agriculture Organization (FAO), these substances, making them relevant to global health, contaminate approximately 25% of all food worldwide. The occurrence of exposure to these mycotoxins is more common in developing countries, where their effects are more harmful to health due to the high rate of malnutrition in these places. The damage caused by them can manifest acutely or chronically, and among them stand out hepatotoxicity, nephrotoxicity, immunogenicity, carcinogenesis, mutagenesis, and teratogenesis and are associated with particularly dangerous nutritional disorders in children from poorer regions. This review focuses on aflatoxins, fumonisins, zearalenone, deoxynivalenol and ochratoxins, with special attention to their impacts on human and animal health, based on experimental studies and case reports. The biomarkers most used in the detection of these substances based on their metabolism are also discussed.

Abstract Dairy cattle when fed on aflatoxin B1 may excrete aflatoxin M1 in milk as a consequence of dietary exposure. Once AFM1 is excreted in milk, it is present in dairy products such as cheese, yogurt, among others. This mycotoxin is quite resistant to temperature therefore heat treatments like pasteurization and ultra-pasteurization are not enough to inactivate it. In this context, this article provides an overview of the biological decontamination methods on milk and cheese of the last decade, as a contribution to evaluate the evolution of this strategy as well as its efficiency according to their authors. Relevant studies published between January 2009 and May 2019 were selected after a systematic search of the literature in PubMed, Science direct and Google Scholar databases. According with our research in the last decade few studies have been published on these methods and unfortunately none of the published studies tested such methods in cheese. Throughout the research many studies on decontamination methods were found, however, in phosphate buffered saline solution or in culture medium. Further studies on biological decontamination on milk and mainly on cheese are necessary for this technique to be better developed and applied to a large scale in the industries.

Abstract Studies conducted in the last decades have demonstrated the probiotic effects of several yeast species. Among the probiotic yeast evaluated, Saccharomyces boulardii is the most common and widely studied species with significant probiotic properties, such as survival capacity in the acidic environment of the gastrointestinal tract and improvements in the intestinal microbiota. In recent years, new evidence indicated that the incorporation of probiotic yeasts is also a promising approach in food development, thus leading to new perspectives for a more integrated strategy for a potential application of S. boulardii in the development of innovative functional food products. In this review, an overview of recent published data on the application of probiotic yeast in foods is presented, especially on the use of S. boulardii as a trend to develop several functional foods, such as non-alcoholic, fermented alcoholic, vegetable, and bakery products.

RESUMO: O objetivo do presente trabalho foi avaliar os efeitos da suplementação com solução de sulfato de cobre (SSC) e arginina em pó (Arg) sobre o desempenho, hormônios tireoidianos e bioquímica sanguínea de frangos de corte alimentados com dietas à base de canola DC. O desenho experimental foi completamente casualizado com fatorial 3 × 3 e nove tratamentos correspondentes a três níveis de inclusão de SSC (0, 125 e 250 mg/kg) e três níveis de Arg (0, 0,1 e 0,2%) (n = 45 para cada tratamento). As rações foram oferecidas ad libitum por 21 dias, de 22 até 42 dias de idade. A eficiência alimentar foi significativamente afetada pela adição de 250 mg/kg de SSC e 0,2% de Arg, assim como pela interação SSC × Arg. A suplementação da DC com SSC melhorou os parâmetros da glândula tireoide e aumentou os níveis plasmáticos de triiodotironina e tiroxina. As aves alimentadas com dietas suplementadas com 0,2% de Arg apresentaram menor nível de glicose sanguínea do que as dos demais tratamentos. A adição de 250 mg/kg de SSC e 0,2% de Arg reduz o estresse causado pelo rápido crescimento dos frangos, além de melhorar as condições gerais de bem estar das aves.

ABSTRACT: The present evaluated the effects of copper sulfate solution (CSS) and arginine powder (Arg) supplements on performance, thyroid hormones and blood biochemistry of broiler chickens fed with canola meal (CM)-based diets. The experimental design was completely randomized with a 3 × 3 factorial and 9 treatments, corresponding to 3 levels of CSS (0, 125 and 250 mg/kg) and 3 levels of Arg (0, 0.1 and 0.2%) (n = 45 per treatment). Feeds were offered ad libitum for 21 days, from 22 to 42 days of age. Feed efficiency was significantly affected by the dietary addition of 250 mg/kg CSS and 0.2% Arg, and by the CSS × Arg interaction. CM supplemented with CSS improved the thyroid gland status and increased the plasma levels of triiodothyronine and thyroxine. Birds fed diets supplemented with 0.2% Arg had lower blood glucose level than the other treatments. The addition of 250 mg/kg CSS and 0.2% Arg reduced the stress caused by the rapid growth of broilers, also increasing the overall bird welfare.

Abstract In this trial, 60 samples of raw cow’s milk collected during summer and winter in five collective centers from Khorasan Razavi Province of Iran were analyzed for penicillin residues, somatic cell counts (SCC), mesophilic counts (TMC) and adulteration materials. The mean penicillin level (0.57 ± 0.20 μg/L) was much lower than the maximum permitted level (MPL) recommended by the Codex Alimentarius Commission (4 μg/L). However, the mean SCC and TMC (Log 5.71 ± 0.08 cells/mL and Log 6.18 ± 0.11 colony forming units (CFU)/mL, respectively) exceeded their respective MPLs adopted in Iran for grade C milk (Log 5.69 cells/mL and Log 6.0 CFU/mL, respectively), with higher counts (P < 0.05) in the summer. High frequencies of samples containing salt (53-60%), detergents (16-20%) and formalin (7-10%) were observed in milk samples, stressing the need for stringent control measures to improve the quality of milk delivered in the evaluated collection centers.

Abstract Silver nanoparticles (AgNPs) are antimicrobial agents that have a wide spectrum of action, including against pathogenic bacteria and spoilage fungi. However, their mechanism of action is not completely clarified. Nowadays, scientific interest on biological synthesis of AgNPs is growing, with emphasis in their extracellular biosynthesis by microbial cells, as it is the most reliable and ecologically correct method for production, yielding no toxic residues. AgNPs may be incorporated to biodegradable and non-biodegradable polymers for the production of food packages with antimicrobial properties, leading to greater safety and longer shelf life. However, it is important to carry out migration tests for new food packages incorporated with AgNPs, based on the effective levels for their inclusion in the packaging materials.

RESUMO: O objetivo do presente trabalho foi verificar a ocorrência de aflatoxinas (AFs) e fumonisinas (FBs) em ingredientes (milho e farelo de soja) e na ração de abate sobre a produtividade de uma empresa integradora de frangos de corte, bem como avaliar seus efeitos sobre produtividade de 20 lotes de frangos produzidos pela empresa e a histologia dos fígados dos frangos após o abate. As amostras de milho apresentaram as maiores frequências de AFs e FBs, em concentrações médias de 29,1 e 2.100µg/kg, respectivamente. As amostras de farelo de soja apresentaram níveis médios de 1,5 e 70µg/kg para AFs e FBs, respectivamente. Os lotes de aves que receberam ração contendo níveis de FBs maiores que 1,000µg/kg apresentaram menor ganho de peso e maior percentual de mortalidade, enquanto que as que receberam ração com AFs iguais ou superiores ao limite de quantificação (LQ) do método analítico apresentaram maior grau de alteração histopatológica no fígado. Houve efeito de diluição de AFs e FBs dos ingredientes, especialmente o milho, à ração no processo de fabricação, porém não suficiente para evitar perdas na produtividade. Os resultados do estudo reforçam a necessidade do controle estrito de micotoxinas no milho destinado à alimentação de frangos de corte.

ABSTRACT: The aim of the present study was to assess the occurrence of aflatoxins (AFs) and fumonisins (FBs) in feed ingredients (corn and soybean meal) and finishing feed in a broiler operation system, as well was to evaluate their effect on the productivity of 20 batches of broilers produced and the histology status of broilers’ liver after slaughter. Corn samples presented the highest frequencies of AFs and FBs, at mean levels of 29.1 and 2,100µg/kg, respectively. Soybean samples presented mean levels of 1.5 and 70µg/kg for AFs and FBs, respectively. Batches of broilers receiving feed containing FB levels higher than 1,000µg/kg had lower weight gain and higher mortality rates, while those fed rations with AFs equal or above the limit of quantification (LOQ) of the analytical method presented higher scores of histological changes in the liver. A dilution effect was observed for AFs and FBs from ingredients, especially corn, to feed during manufacture, whilst not enough to prevent losses in productivity. Results of this trial highlighted the need for strict control of mycotoxins in corn intended for broilers.

Resumo Folhas de oliveira contêm oleuropeína, um composto fenólico conhecido por seu potencial antioxidante, antimicrobiano, anti-inflamatório, entre outros. Este trabalho teve como objetivo estudar a extração da oleuropeína, utilizando solvente hidroalcoólico. Folhas de oliveira (FO) foram maceradas a 25 °C com um solvente (S), formado por uma mistura de etanol e água (70:30 v/v), nas proporções FO:S iguais a 1:8, 1:6 e 1:3, em massa, com ou sem a presença de ácido acético. Observou-se que o maior teor de oleuropeína no extrato liofilizado (18 g/100 g, aproximadamente) foi obtido no procedimento utilizando a razão FO:S = 1:3, com ácido acético. Por fim, foi realizado um teste para verificar o efeito do extrato liofilizado sobre o tempo de indução em amostras de azeite de oliva extravirgem e refinado, verificando-se um aumento de três e duas horas, respectivamente. Concluiu-se que é possível obter extratos de folhas de oliveira utilizando solventes renováveis e que a oleuropeína pode atuar como um antioxidante natural no azeite, melhorando sua estabilidade oxidativa.

Abstract Olive leaves contain oleuropein, a substance known for its antioxidant, antimicrobial and antiinflammatory potentials, amongst others. The objective of this work was to study the extraction of oleuropein using a hydroalcoholic solvent. Olive leaves (OL) were macerated at 25 °C with a solvent (S) consisting of a mixture of ethanol and water (70:30 v/v), with OL:S mass ratios equal to 1:8, 1:6 and 1:3, with or without the presence of acetic acid. The highest oleuropein content found in the freeze dried extract (approximately 18 g/100 g) was obtained from the procedure using an OL:S ratio = 1: 3, in the presence of acetic acid. Finally, a test was carried out to verify the effect of the freeze dried extract on the induction time in samples of extra-virgin and refined olive oil, and increases of 3 and 2 hours, respectively, were observed. It was concluded that it is possible to obtain olive leaf extracts using renewable solvents, and that oleuropein can act as a natural antioxidant in olive oil, improving its oxidative stability.

ABSTRACT This study aimed to investigate the effect of peracetic acid (PAA, 0.5%) on adherent cells of three strains of Listeria monocytogenes strains belonging to serotypes 4b and 1/2b that had been previously isolated from the environment of a Brazilian cheese plant. The assays were conducted using polystyrene microplates and stainless steel coupons and the adhered cells were treated with PAA for 60, 120 and 180 s. On stainless steel, biofilms were partially inactivated by PAA after 60 s and almost 100% of the cells were damaged within 180 s using epifluorescence microscopy with LIVE/DEAD® staining. On polystyrene microplates, PAA decreased (P<0.05) biofilm biomass produced by the three L. monocytogenes isolates at 60 s, when compared with controls (no PAA treatment). However, PAA did not completely eliminate L. monocytogenes cells on polystyrene microplates (decreasing 1.8-2.5 log cycles after treatment with PAA for 180 s). The correct concentration and contact time of PAA is critical for eliminating biofilms formed by L. monocytogenes on stainless steel surfaces, although further studies are needed for defining efficient PAA treatments to remove adherent cells of this pathogen on plastic polymers.

RESUMO: Neste trabalho foram determinados os níveis de ácido fólico e de fumonisina B1 (FB1) em farinha de milho consumida por 24 voluntários residentes em um campus universitário no estado de São Paulo, bem como sua relação com as concentrações de ácido fólico sérico nos indivíduos. As análises de ácido fólico e de FB1 em farinha de milho foram realizadas por cromatografia líquida de alta eficiência (CLAE), enquanto a determinação de ácido fólico sérico foi feita por kit de imunoensaio. Detectou-se a FB1 em 100% das amostras de farinha de milho, em níveis que variaram de 142 a 3.037 µg kg-1 (média: 738 ± 591 µg kg-1). As concentrações de ácido fólico nas amostras de farinha de milho ficaram entre < 0,3 µg kg-1 (limite de quantificação) e 1.705 µg kg-1, com média de 713 ± 435 µg kg-1, o que representa 47% do limite mínimo exigido pela Agência Nacional de Vigilância Sanitária (ANVISA) para farinhas de milho comercialmente disponíveis. Nas amostras de soro humano, os níveis de ácido fólico variaram de 6,7 a 24,0 ng mL-1 (média: 13,4 ± 5,4 ng mL-1). Não houve correlação (p < 0,05) entre os níveis de ácido fólico no soro dos indivíduos e as concentrações de FB1 ou ácido fólico nas amostras de farinha de milho. Outros estudos são necessários para estimar a ingestão total de FB1 por meio da dieta para averiguar os efeitos das fumonisinas sobre a absorção de ácido fólico nos indivíduos avaliados.

ABSTRACT: In the present study, folic acid and fumonisin B1 (FB1) levels were determined in corn flour consumed by 24 volunteers, residents in a university campus in São Paulo State, as well as its relationship with folic acid in serum of individuals. Analyses of folic acid and FB1 in corn flour were performed by high-performance liquid chromatography (HPLC), while the determination of folic acid in serum was accomplished using an immunoassay kit. FB1 was detected in 100% of corn samples, at levels ranging from 142 to 3,037 µg kg-1 (which means: 738 ± 591 µg kg-1). The concentrations of folic acid in corn flour samples ranged from < 0.3 µg kg-1 (limit of quantification) to 1,705 µg kg-1, with a mean of 713 ± 435 µg kg-1, which represents 47% of the minimum required by National Agency of Health Surveillance (ANVISA) for corn flour commercially available. The levels of folic acid in human serum samples ranged from 6.7 to 24.0 ng mL-1 (meaning: 13.4 ± 5.4 ng mL-1). No correlations were observed (p < 0.05) between the folic acid levels in serum of individuals and the concentrations of FB1 or folic acid in corn flour samples. Further studies are needed to estimate the total intake of FB1 in the diet to assess the effects fumonisins on the absorption of folic acid in the individuals evaluated.

This study aimed to verify the in vitro ability of beer fermentation residue (BFR) containing Saccharomyces cerevisiae cells and five commercial products that differed in the viability and integrity of S. cerevisiae cells to remove aflatoxin B1 (AFB1) from a citrate-phosphate buffer solution (CPBS). BFR was collected at a microbrewery and prepared by drying and milling. The commercial yeast-based products were as follows: inactive intact yeast cells from beer alcoholic fermentation, inactive intact yeast cells from sugarcane alcoholic fermentation, hydrolyzed yeast cells, yeast cell walls and active yeast cells. Adsorption assays were performed in CPBS spiked with 1.0 μg AFB1/mL at pH 3.0 and 6.0 for a contact time of 60 min at room temperature. Analysis of AFB1 in the samples was performed by high performance liquid chromatography. AFB1 adsorption by the products ranged from 45.5% to 69.4% at pH 3.0 and from 24.0% to 63.8% at pH 6.0. The higher percentages (p < 0.05) of AFB1 binding at both pH values were achieved with products containing hydrolyzed yeast cells or yeast cell walls rather than intact cells. The AFB1 binding percentages of BFR were 55.0 ± 5.0% at pH 3.0 and 49.2 ± 4.5% at pH 6.0, which was not significantly different (p > 0.05) from commercial products containing inactive intact yeast cells. The results of this trial indicate that the yeast-based products tested, especially the BFR, have potential applications in animal feeds as a suitable biological method for reducing the adverse effects of aflatoxins.

This study aimed to compare Lactobacillus rhamnosus growth in MRS (de Man, Rogosa and Sharpe) broth and a culture medium containing milk whey (MMW) and to evaluate aflatoxin B1 (AFB1) adsorption capacity by bacterial cells produced in both culture media. L. rhamnosus cells were cultivated in MRS broth and MMW (37 °C, 24 hours), and bacterial cell concentration was determined spectrophotometrically at 600 nm. AFB1 (1 µg/ml) adsorption assays were conducted using 1 x 10(10) non-viable L. rhamnosus cells (121 °C, 15 minutes) at pHs 3.0 and 6.0 and contact time of 60 minutes. AFB1 quantification was performed by High Performance Liquid Chromatography. Bacterial cell concentration in MMW was higher (9.84 log CFU/ml) than that in MRS broth (9.63 log CFU/ml). There were no significant differences between AFB1 binding results at the same pH value (3.0 or 6.0) for the cells cultivated in MRS broth (46.0% and 35.8%, respectively) and in MMW (43.7% and 25.8%, respectively), showing that MMW can adequately replace the MRS broth. Therefore, it can be concluded that the use of L. rhamnosus cells cultivated in MMW offers advantages such as reduction in large scale production costs, improvement of environmental sustainability, and being a practicable alternative for decontamination of food products susceptible to aflatoxin contamination.

As aflatoxinas são responsáveis por grandes prejuízos à saúde de aves comerciais e à produção animal, essencialmente por perda financeira decorrente de diminuição de ganho de peso dos animais. A detoxificação da ração ou de suas matérias-primas, bem como tentativas de diminuir a contaminação fúngica que leva à produção das toxinas, além de técnicas nutricionais, são métodos que podem ser adotados para tentar diminuir os efeitos deletérios sobre os frangos de corte. Sendo assim, o objetivo desta revisão é mostrar o que a ciência fornece como meios de detoxificação e/ou redução de contaminação por fungos; e algumas substâncias naturais que auxiliam a produção avícola na redução de micotoxicoses .

Aflatoxins are answerable for large losses for the health of commercial birds and for the animal production, essentially by financial losses due to animal body weight diminution. Detoxification of feed or its raw materials, well as attempts of diminution of fungal contamination that leads to toxins production, beyond of nutritional techniques, are methods that can be adopted trying to diminish deleterious effects among broiler chickens. This way, the objective of this review is to show what science supplies like ways of detoxification and/or reduction of contamination by fungi, and some natural substances that help poultry production on reduce mycotoxicosis.

Este estudo teve como objetivo correlacionar a atividade da plasmina e as concentrações de frações de caseína no leite longa vida (UAT) produzido em usina de beneficiamento do Estado de São Paulo. O leite cru foi submetido à pasteurização (72-76 ºC, 15-20 segundos), seguida da esterilização por processo de injeção direta de vapor (132-136 ºC, 34 segundos), sendo homogeneizado e envasado assepticamente em embalagens cartonadas. Quatro lotes de UAT foram analisados para atividade de plasmina e frações de caseína após 10, 30, 60, 90 e 120 dias de armazenamento. A atividade de plasmina no leite longa vida aumentou durante o armazenamento, correlacionando-se negativamente com a concentração de αs1-caseína. A atividade da plasmina apresenta resistência à esterilização do leite pelo processo UAT e aumenta a proteólise do leite longa vida como consequência da degradação da αs1-caseína.

The objective of this study was to correlate the plasmin activity and the concentrations of casein fractions in ultra-high-temperature (UHT) milk manufactured in a dairy plant in the State of Sao Paulo, Brazil. Raw milk was pasteurized (72-76 ºC, 15-20 seconds), followed by direct steam injection into the milk (132-136 ºC, 34 seconds), then homogenized and aseptically packaged into cartons. Four replicates of the UHT milk were analyzed for the plasmin activity and casein fractions after 10, 30, 60, 90 and 120 days of storage. Plasmin activity significantly increased in the UHT milk during storage and was negatively correlated with the concentration of αs1-casein. Plasmin is resistant to milk sterilization by the UHT process, and increases proteolysis of the UHT milk as a consequence of αs1-casein degradation.