Assessment of mycotoxin exposure and risk characterization using occurrence data in foods and urinary biomarkers in Brazil.

This study aimed to assess the exposure of Brazilian residents (N = 86) from rural areas to multiple mycotoxins and characterize the associated risk in two sampling periods (SP) (April-May and December/2016). Mycotoxins in food and urine samples were determined by liquid chromatography coupled to tandem mass spectrometry. Mean probable daily intake (PDI) values based on occurrence data in foods in both SP varied from 0.007 to 0.013, 0.069 to 1.002, 0.119 to 0.321 and 0.013-0.156 µg kg-1 body weight (bw) day-1 for aflatoxins (AFs), deoxynivalenol (DON), fumonisins (FBs) and zearalenone (ZEN), respectively. Mean PDI values based on urinary biomarkers were 0.001, 84.914, 0.031, 0.377 and 0.002 µg kg-1 bw day-1 for AFB1, DON, ochratoxin A (OTA), FB1 and ZEN, respectively. Hazard quotient (HQ) calculated using food data revealed a potential health concern for ZEN in 2nd SP. HQ > 1 based on urinary biomarkers were observed for DON in the two SP. Although OTA was not detected in any food sample, the HQ based on urinary OTA levels was >1 in the 1st SP. Margin of exposure values for AF from food and urine data in the 1st SP were below 10,000, indicating potential health risks.

Co-occurrence of mycotoxins in maize food and maize-based feed from small-scale farms in Brazil: a pilot study.

A limited survey was conducted to assess the co-occurrence of aflatoxins (AF) B1, B2, G1, and G2; fumonisins (FB) B1 and B2; ochratoxin A (OTA); zearalenone (ZEN); and deoxynivalenol (DON) in maize food (N = 26) and animal feed (N = 45) collected from 21 small-scale farms from the states of São Paulo (SP) and Santa Catarina (SC), Brazil. Samples evaluated were maize meal and maize flour for human consumption available in the farm households, and maize-based feed intended for broiler chicks, laying hens, and dairy cows. Analyses of mycotoxins were performed by ultra-performance liquid chromatography coupled with tandem mass spectrometry. The median levels of mycotoxins found in maize food were 2.5 µg/kg (total AF), 120 µg/kg (total FB), 13 µg/kg (ZEN), and 57 µg/kg (DON). All values were below the Brazilian tolerance limits, except for total FB in one sample of maize flour. In feed samples, median levels of total AF, total FB, ZEN, and DON were 100 µg/kg, 680 µg/kg, 160 µg/kg, and 200 µg/kg, respectively. The co-occurrence of two or more mycotoxins was confirmed in 35% and 51% of maize food and feed, respectively. Results indicate a low human exposure to mycotoxins in the small-scale farms evaluated and a higher exposure of farm animals to mycotoxins in the feed.

Toxicological effects of fumonisin B1 in combination with other Fusarium toxins.

Fusarium is a fungal genus spread worldwide commonly associated to the production of several mycotoxins, where fumonisins (FBs) are of major importance due to its prevalence. Since mycotoxins have been reported to cause deleterious effects on mammalians, including carcinogenic, neurotoxic, estrogenic, and immune-suppressive, many countries had established regulations on the tolerated concentrations of such substances in foods and animal feed. Even though many mycotoxins - especially fusariotoxins - are concomitantly found in a single matrix, there is no regulation on co-occurrence levels. This is possibly a result of the lack of data in the literature on the toxicological interactions between different mycotoxins. Considering this, it is of utmost importance to gather what is currently known about the combination of FBs, considered to be the most ubiquitous mycotoxins, with other frequently reported fusariotoxins, such as zearalenone (ZEA), deoxynivalenol (DON), nivalenol (NIV), T-2 toxin (T-2), and other emerging mycotoxins. This paper gives an overview about the toxic effects of fusariotoxins individually and combined to FB1, also gathering the mechanisms and probable interactions between them. This important information may help to develop regulations covering multi-mycotoxins contamination, a growing concern of current days.

Combination of garlic essential oil, allyl isothiocyanate, and nisin Z as bio-preservatives in fresh sausage.

The effects of natural antimicrobial compounds (garlic essential oil [GO], allyl isothiocyanate [AITC], and nisin Z [NI]) on microbiological, physicochemical and sensory characteristics of fresh sausage were assessed. The minimum inhibitory concentrations (MICs) and the minimum bactericidal concentrations (MBCs) towards Escherichia coli O157:H7 and Lactobacillus plantarum were determined in vitro. Sausages inoculated with E. coli O157:H7, were treated with different combinations of antimicrobials and assessed for microbiological and physicochemical parameters during storage (6C for 20 d). Treatments that presented the greatest antimicrobial effects were subjected to sensory evaluation. Combinations of 20 mg/kg NI + 125 µL/kg GO + 62.5 µL/kg AITC or 20 mg/kg NI + 62.5 µL/kg GO + 125 µL/kg AITC were effective in reducing E. coli O157H7 and spoilage lactic acid bacteria, and maintained the physicochemical characteristics of fresh sausage. Combinations of NI, GO and AITC were effective to improve the safety and the shelf life of fresh sausage, with no impact on its sensory acceptance.

Evaluation of gaseous allyl isothiocyanate against the growth of mycotoxigenic fungi and mycotoxin production in corn stored for 6 months.

BACKGROUND: Brazil produces approximately 63 million tons of corn kernels annually, which is commonly contaminated with fungi and mycotoxins. The objective of this study was to evaluate the efficacy of gaseous allyl isothiocyanate (AITC) to inhibit the growth of Aspergillus parasiticus and Fusarium verticillioides, and mycotoxin production (aflatoxins B1 , B2 , G1 and G2 , fumonisins B1 and B2 ) in corn during 180 days of storage. RESULTS: AITC at 50 µL L-1 resulted in a significant reduction of the fungal population (P < 0.05) after 180 days, decreasing 3.17 log(CFU g-1 ) and 3.9 log(CFU g-1 ) of A. parasiticus and F. verticillioides respectively in comparison with the control. In addition, 10 and 50 µL L-1 treatments prevented the production of fumonisin B1 for the whole period. Aflatoxins were not detected in either control or treated groups. Residual levels of AITC in corn treated with 10 µL L-1 and 50 µL L-1 were detected up to 14 days and 30 days respectively. CONCLUSION: Prophylactic treatment with AITC reduced the fungal population and inhibited fumonisin B1 production in stored corn, exhibiting great potential to be applied in corn silos to prevent fungi contamination and minimize mycotoxin levels. © 2018 Society of Chemical Industry.

Fumigation of Brazil nuts with allyl isothiocyanate to inhibit the growth of Aspergillus parasiticus and aflatoxin production.

BACKGROUND: Brazil produces approximately 40 000 tons of Brazil nuts annually, which is commonly contaminated with fungi and mycotoxins. Gaseous allyl isothiocyanate (AITC) was used to inhibit the growth of Aspergillus parasiticus and its production of aflatoxins (AFs) in Brazil nuts. RESULTS: Nuts were inoculated with 104 spores g-1 of A. parasiticus and placed in airtight glass jars with controlled relative humidity (RH = 95 or 85%). Samples were treated with 0, 0.5, 1.0 or 2.5 µL L-1 of gaseous AITC and analyzed after 30 days to determine the fungal population and AFs content. Samples were also submitted to sensory evaluation. AITC at 2.5 µL L-1 could completely inhibit the fungal growth and AFs production in both the RH tested. AITC at 0.5 and 1 µL L-1 did not affect the microbial growth at RH = 95%, but 1 µL L-1 reduced the production of AFs by ∼50%. All AITC treatments reduced the fungal population and AFs to undetectable levels at RH = 85%. None of the concentrations altered sensory characteristics of Brazil nuts. CONCLUSION: Gaseous AITC could be used as an alternative to inhibit the growth of A. parasiticus during storage and transport of Brazil nuts. © 2017 Society of Chemical Industry.

Combination of essential oil compounds and phenolic acids against Escherichia coli O157:H7 in vitro and in dry-fermented sausage production.

Escherichia coli O157:H7 is a foodborne pathogen that causes hemorrhagic colitis and hemolytic uremic syndrome. The low dose of infection and severity of the disease represent a concern to public health. Natural compounds have been widely applied as food additives to replace synthetic preservatives. The aim of this study was to determine the efficiency of essential oil compounds (EOCs) in combination with phenolic acids (PA) in vitro and in dry-fermented sausage production. Minimum Inhibitory Concentration (MIC) and Fractional Inhibitory Concentration index (FICindex) were determined for a 5-strain mixture of E. coli O157:H7. Batches of sausage tainted with E. coli O157:H7 were produced using Pediococcus pentosaceus UM 116P and Staphylococcus carnosus UM 123M as starter cultures. The best combination of EOCs and PAs found in vitro was used as an additive. Chemical-physical and microbiological analyses were evaluated weekly from day 0 to 35 after production. Sensory evaluation (texture, odor, flavor, appearance and overall evaluation) of E. coli-free sausages was conducted using a 9-point hedonic scale with 56 untrained volunteers. The MIC values of allyl isothiocyanate (AITC), carvacrol (CAR), ferulic acid (FA), o-coumaric acid (CA) and p-hydroxybenzoic acid (AHB) were, respectively, 0.25; 1.3; 5.12; 18.27; and 37mM. AITC combined with CA had a synergistic effect (FICindex=0.25) and together they were applied in the production of dry fermented sausage at concentrations of 10× FIC and 20× FIC. Aw had no significant difference among treatments, whereas the pH of 10× FIC and 20× FIC were higher than the control. E. coli O157:H7 was reduced by >5logCFU/g with 20× FIC after 21d, and by 2.8logCFU/g with 10× FIC after 35d. Sensory analysis showed that the combination of AITC and ο-coumaric acid in both treatments presented lower scores in the 5 categories when compared to the control, but none of the parameters received a negative score. This study demonstrated that the combination of AITC and ο-coumaric acid at 20× FIC reduced E. coli O157:H7 in compliance with the North American legislation, but adjustments in the dose are necessary to improve the sensory characteristics of the final product.

Determination of Aflatoxin B1-Lysine in Pig Serum and Plasma by Liquid Chromatography-Tandem Mass Spectrometry.

Aflatoxin B1 (AFB1) is a hepatocarcinogen produced by certain Aspergillus species growing on crops. After biotransformation in the liver, AFB1 generates several metabolites, one of which is AFB1 bound to lysine on serum albumin. AFB1-lysine (AFB1-lys) is a digest product of AFB1-albumin and is considered a biomarker of exposure to AFB1 in humans and animals. The objectives of this paper were to evaluate the performance characteristics of a new analytical method for determination of AFB1-lys levels in pig serum, heparinized and ethylenediaminetetraacetic acid (EDTA) plasma and to evaluate the interference of these anticoagulants in AFB1-lys quantification. Blank blood samples were obtained from eight crossbreed 91-day-old barrows fed AFB1-free diets. Pooled samples (n = 3) and individual samples of serum, EDTA and heparinized plasma collected from five pigs were enzymatically digested with pronase at 37°C for 4 h. AFB1-lys was isolated by solid-phase extraction and quantified by liquid chromatography coupled to tandem mass spectrometry. The analytical method was applied for determination of AFB1-lys in serum and EDTA plasma collected from five 49-day-old crossbreed barrows fed ad libitum diets containing 1.1 mg of AFB1 per kg of feed during 7 days (three animals) or 42 days (two animals). Samples of heparinized plasma were only available from animals intoxicated for 42 days. All animals had lower levels of AFB1-lys in EDTA plasma samples (24.78-37.40 ng/mL), when compared to serum (49.32-252.07 ng/mL-1) or heparinized plasma (176.81 and 264.24 ng/mL-1). EDTA did not interfere in AFB1-lys standard detection, but our findings suggest that EDTA should be avoided during blood collection since it affects the pronase activity in AFB1-albumin adduct digestion and, consequently, causes a reduction in the AFB1-lys levels. Hence, determination of AFB1-lys in serum and heparinized plasma is an approach to assess an individual's exposure of swine to AFB1.

Relação entre níveis de fumonisina B1 e ácido fólico em farinha de milho e a concentração de ácido fólico no soro humano / Relationship between fumonisin B1 and folic acid levels in foods and the folic acid concentration in human serum

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.(AU)

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.(AU)

Ganho de peso, consumo de ração e histologia de órgãos de leitões alimentados com rações contendo baixos níveis de fumonisina B1 / Weight gain, feed consumption and histology of organs from piglets fed rations containing low levels of fumonisin B1

A fumonisina B1 (FB1) é um metabólito secundário produzido principalmente por Fusarium verticilioides em diversos tipos de alimentos, principalmente o milho, o qual constitui a base para composição de rações para várias espécies de animais domésticos. A FB1é particularmente tóxica para suínos, cujas manifestações clínicas são evidentes em animais expostos a altas concentrações de FB1 na ração (em geral, acima de 30mg/kg). No entanto, são escassos os estudos sobre os efeitos da FB1em suínos alimentados com rações contendo baixas concentrações de fumonisinas, as quais são mais prováveis de serem encontradas em condições de campo. O objetivo do estudo foi avaliar os efeitos da exposição de leitões a baixos níveis de FB1 na ração, durante 28 dias, sobre o ganho de peso, consumo de ração, peso relativo de órgãos e aspectos histológicos do baço, fígado, pulmões, rins e coração. Vinte e quatro leitões foram distribuídos em 4 grupos experimentais e alimentados com rações contendo 0mg (controle), 3,0mg, 6,0mg ou 9,0mg FB1/kg de ração. As diferentes dietas não afetaram (P>0,05) o ganho de peso e nem o peso relativo dos órgãos analisados. Não foram constatadas lesões macroscópicas ou histopatológicas no baço, fígado, rins e coração. No entanto, foram observadas lesões histopatológicas nos pulmões de todos os suínos alimentados com rações contaminadas com fumonisinas, indicando que nenhum dos níveis de FB1 usados no experimento poderia ser considerado como seguro para suínos. São necessários novos estudos sobre os mecanismos de ação tóxica da FB1 em suínos, sobretudo em condições de exposição prolongada a baixos níveis de contaminação na ração.

Fumonisin B1 (FB1) is a secondary metabolite produced mainly by Fusarium verticilioides in several types of foods, particularly corn, which is the basis for composition of feed for several domestic animals. FB1 is particularly toxic to pigs, being the clinical manifestations evident in animals exposed to high concentrations of FB1 in the diet (generally above 30mg/kg). However, there are few studies on the effects of FB1 on pigs fed rations containing low concentrations of fumonisin, which are most probably found under field conditions. The aim of the study was to evaluate the effects of a 28-day exposure of piglets to low levels of FB1 in the feed on the weight gain, feed consumption, organ weights and histological aspects of the spleen, liver, lungs, kidneys and heart. Twenty-four pigs were assigned into 4 experimental groups and fed diets containing 0mg (control), 3.0mg, 6.0mg or 9.0mg FB1/kg diet. The different diets did not affect (P>0.05) the weight gain or the weight of organs examined. There were no macroscopic or histological lesions in the spleen, liver, kidneys and heart. However, histological lesions were found in the lungs from all animals fed rations containing fumonisin, hence indicating that none of the FB1 levels used in the experiment could be considered as safe for piglets. Further studies on the mechanisms of toxic action of FB1 in pigs are needed, particularly under conditions of prolonged exposure to low contamination levels in the diet.

Changes in food caused by deep fat frying--a review.

Deep-fat frying is widely used in food industries because of its low cost and high demand, since it produces convenient food of high acceptability. The process is based on the oil-food interaction at high temperatures, which cooks and dehydrates the food, leading to physical and chemical changes, such as starch gelatinization, protein denaturation, flavoring and color production via Maillard reaction. Some food and oil compounds are lost in the frying process, and potentially toxic compounds are developed in the oxidized oil. Although widely studied, many of these compounds have not been fully identified. The purpose of this study was to review literature findings on changes in food caused by the frying process.

Changes in food caused by deep fat frying - A review

Deep-fat frying is widely used in food industries because of its low cost and high demand, since it produces convenient food of high acceptability. The process is based on the oil-food interaction at high temperatures, which cooks and dehydrates the food, leading to physical and chemical changes, such as starch gelatinization, protein denaturation, flavoring and color production via Maillard reaction. Some food and oil compounds are lost in the frying process, and potentially toxic compounds are developed in the oxidized oil. Although widely studied, many of these compounds have not been fully identified. The purpose of this study was to review literature findings on changes in food caused by the frying process.

Alterações nos alimentos causados pelo processo de fritura- Revisão. O processo de fritura é amplamente utilizado em indústria de alimentos devido ao baixo custo e alta demanda pela praticidade e grande aceitação. O processo é baseado na interação óleo-alimento a altas temperaturas, que cozinha o alimento e desidrata, levando a alterações físicas e químicas assim como a gelatinização do amido, desnaturação de proteínas, aroma e produção de cor através da reação de Maillard. Alguns compostos presentes nos alimentos e no óleo são perdidos no processo de fritura, e componentes potencialmente tóxicos são desenvolvidos nos óleos oxidados. Embora diversos tenha havido avanços na identificação desses componentes, muitos ainda não foram identificados. A proposta desse trabalho foi avaliar as alterações nos alimentos causadas pelo processo de fritura.