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Mycotoxins are toxic metabolites naturally produced by some fungi species, which cause severe health effects in humans and animals. Although contaminated foods are major routes of human exposure to...

The current investigation was aimed to estimate the prevalence and concentration of ochratoxin A (OTA) in different types of coffee and coffee-based products with the aid of a systematic review and meta-analysis. Therefore, the recommended databases including PubMed, Scopus, and Embase from Jan 1983 to Oct 2018 were screened to retrieve the related citations. In this regard, among 1041 explored articles in the identification step, thirty six articles with 3182 samples were included in the meta-analysis and meta-regression. According to findings, the global pooled concentration and prevalence of OTA was calculated as 3.21 μg/kg (95% CI: 3.08-3.34 μg/kg) and 53.0 % (95% CI: 43.0-62.0), respectively. Also, direct correlations between the increases in poverty as well as the amount of annual precipitation and prevalence of OTA was noted, while with decreasing in HDI the prevalence of OTA in coffee significantly was increased. Moreover, the lowest and highest concentrations of OTA in coffee were observed in Taiwan (0.35 μg/kg) and Turkey (79.0 μg/kg), respectively. The outcome of this meta-analysis can be used for the building of risk assessment models aiming to derive data for the development of specific actions to reduce the exposure to this mycotoxin in coffee and coffee-based products.

This study aimed to assess the exposure of 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. Highlights: Mycotoxin exposure evaluation and risk assessment in rural areas from Brazil; 53% of food samples analyzed (N=203) presented at least one type of mycotoxin; Urinary biomarkers of mycotoxins found at levels of 0.02–12.0 ng mg−1 creatinine; PDI based on biomarkers in urine exceed recommended TDI values for DON and OTA; Margin of exposure values indicated health risks associated with dietary aflatoxins. The authors would like to thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) – grant no. 400649/2014-4, for financial support and fellowships. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001. Paula Alvito and Ricardo Assunção acknowledge the support from National Institutes of Health, Dr. Ricardo Jorge, and CESAM by means of Fundação para a Ciência e a Tecnologia [UID/AMB/50017/2013], through national funds, and the co-funding by the FEDER [POCI-01- 0145-FEDER-00763], within the PT2020 Partnership. info:eu-repo/semantics/publishedVersion

The increased consumption of plant-based foods has intensified the concern related to mycotoxin intoxication. This study aimed to investigate the effect of selected lactic acid bacteria (LAB) strains on the growth of Aspergillus parasiticus NRRL 2999 and its production of aflatoxin (AF). The ability of the heat-killed (100°C for 1 h) LAB strains to bind aflatoxin M1 (AFM1) in milk and aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN) in potassium phosphate buffer (PPB) was also evaluated in vitro. Ten LAB strains were tested individually, by inoculating them simultaneously with the fungus or after incubation of the fungus for 24 or 48 h at 25°C. Double layer yeast extract sucrose (YES) agar, de Man Rogosa and Sharpe (MRS) agar, and YES broth were incubated for 7 days at 25°C to follow the development of the fungus. Levilactobacillus spp. 3QB398 and Levilactobacillus brevis 2QB422 strains were able to delay the growth of A. parasiticus in YES broth, even when these strains were inoculated 24 h after the fungus. The inhibitory effect of these LAB strains was confirmed by the reduction of fungus colony size, suggesting dominance of LAB by competition (a Lotka-Voltera effect). The production of AFB1 by A. parasiticus was inhibited when the fungus was inoculated simultaneously with Lactiplantibacillus plantarum 3QB361 or L. plantarum 3QB350. No AFB1 was found when Levilactobacillus spp. 2QB383 was present, even when the LAB was inoculated 48 h after the fungus. In binding studies, seven inactivated LAB strains were able to promote a reduction of at least 50% the level of AFB1, OTA, and ZEN. This reduction varied depending on the pH of the PPB. In milk, however, only two inactivated LAB strains were able to reduce AFM1, with a reduction of 33 and 45% for Levilactobacillus spp. 3QB398 (Levilactobacillus spp.) and L. brevis 2QB422, respectively. Nevertheless, these results clearly indicate the potential of using LAB for mycotoxin reduction.

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 Sao 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.

Mycotoxins are secondary metabolites produced by fungi that occur naturally in foodstuffs, which can cause a large variety of toxic effects on vertebrates including humans. The objectives of this work were to evaluate the co-occurrence of 11 mycotoxins in food products, feed for broiler chicks, laying hens and dairy cattle, assess the human exposure to mycotoxins through food analysis versus consumption data and multimycotoxin biomarkers in urine, and characterize the associated risk of mycotoxin exposure in Brazilian rural areas. Sampling procedures were conducted in 38 small-scale dairy and poultry farms in the surroundings of Pirassununga and Descalvado (State of São Paulo), Pinhalzinho and Erval Velho (State of Santa Catarina). In these farms a total of 86 volunteers were recruited and instructed to provide samples of the morning urine (N = 162) in two sampling periods (April-May/2016 and December/2016), along with samples of rice (N = 66), bean (N = 59), wheat (N = 39), corn flour (N = 21) and corn meal (N = 18) available in their households. Samples of feed for broilers (N = 10), laying hens (N = 20) and dairy cattle (N = 15) were also collected. All samples were analyzed by ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-MS/MS) for determination of aflatoxins (AF) B1, B2, G1 and G2, fumonisins (F) B1 and B2, ochratoxin A (OTA), zearalenone (ZEN), deoxynivalenol (DON), toxin T-2 and toxin HT-2 in food products and feeds, and AFM1, AFP1, AFQ1, FB1, OTA, T-2, HT-2, DON, de-epoxideoxynivalenol (DOM-1), ZEN, α-zearalenol (α-ZEL), β-zearalenol (β-ZEL) and 15-acetyl-DON in urine samples. The mycotoxin levels in urine were adjusted to creatinine concentration in each sample analyzed. 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 51% of feed samples. Results indicate a high exposure of farm animals to mycotoxins in the feed, hence emphasizing the need to improve the feed quality regarding the contamination with mycotoxins in small-scale farms in Brazil, and the necessity of include feed in Brazilian regulation, especially for AF, FB, and ZEN. Mycotoxin levels above the Brazilian maximum permitted levels (MPL) were found in rice (1.5%), wheat flour (12.8%) and corn flour (14.3%) samples. Urine determinations revealed the presence of AFM1 and AFP1, DON, OTA, FB1 and ZEN at levels of 0.02-12.0 ng/mg creatinine. Regarding the probable daily intake (PDI) based on food data, only ZEN (0.156 µg/kg b.w./day) had a Hazard Quotient (HQ) above the tolerance (> 1). PDI values based on urinary levels for DON, OTA and total AF were 84.914, 0.031 and 0.001 µg/kg b.w./day, respectively, resulting in HQ values > 1, which may indicate health risks for the population studied. An informal intervention by means of educational activities and delivery of an information flyer during the first sampling period did not change the exposure levels to mycotoxins in the second sampling period. Further studies are needed to identify food items other than those analyzed in this work as sources of dietary mycotoxins, as well as the contribution of inhalation of contaminated dusts for the exposure. This is the first study to report the risk assessment of mycotoxins based on food and urinary levels in rural areas in Brazil. As micotoxinas são metabólitos secundários produzidos por fungos que ocorrem naturalmente em alimentos, das quais podem causar uma grande variedade de efeitos tóxicos em vertebrados, incluindo humanos. Os objetivos deste trabalho foram avaliar a co-ocorrência de 11 micotoxinas em alimentos, rações para frangos de corte, poedeiras e gado leiteiro, avaliar a exposição humana a micotoxinas através de análise de alimentos versus dados de consumo e biomarcadores de múltiplas micotoxinas na urina, e caracterizar o risco associado de exposição a micotoxinas em áreas rurais brasileiras. Os procedimentos de amostragem foram conduzidos em 38 propriedades leiteiras e avícolas de pequeno porte nos arredores de Pirassununga e Descalvado (SP), Pinhalzinho e Erval Velho (SC). Nestas fazendas, um total de 86 voluntários foram recrutados e instruídos a fornecer amostras da primeira urina da manhã (N = 162) em dois períodos de amostragem (abril-maio/2016 e dezembro/2016), juntamente com amostras de arroz (N = 66) , feijão (N = 59), trigo (N = 39), farinha de milho (N = 21) e fubá (N = 18) disponíveis em suas residências. Amostras de ração para frangos de corte (N = 10), poedeiras (N = 20) e bovinos leiteiros (N = 15) também foram coletadas. Todas as amostras foram analisadas por cromatografia líquida de ultra-performance acoplada a espectrometria de massas (UPLC-MS/MS) para determinação de aflatoxinas (AF) B1, B2, G1 e G2, fumonisina (FB) B1 e B2, ocratoxina A (OTA), zearalenona (ZEN), desoxinivalenol (DON), toxina T-2 e toxina HT-2 em produtos alimentícios e rações, e AFM1, AFP1, AFQ1, FB1, OTA, T-2, HT-2, DON, de-epóxido-oxinivalenol (DOM-1), ZEN, α-zearalenol (α-ZEL), β-zearalenol (β-ZEL) e 15-acetil-DON em amostras de urina. Os níveis de micotoxinas na urina foram ajustados à concentração de creatinina em cada amostra analisada. Em amostras de ração, os níveis médios de AF total, FB total, ZEN e DON foram de 100 µg/kg, 680 µg/kg, 160 µg/kg e 200 µg/kg, respectivamente. A co-ocorrência de duas ou mais micotoxinas foi confirmada em 51% das amostras de ração. Os resultados indicam uma alta exposição de animais de fazenda à micotoxinas na ração, enfatizando a necessidade de melhorar a qualidade das rações em fazendas de pequena escala no Brasil, referente as micotoxinas, e a necessidade de incluir ração na legislação brasileira, especialmente para AF, FB e ZEN. Os níveis de micotoxinas acima dos níveis máximos permitidos no Brasil (LMP) foram encontrados em arroz (1,5%), farinha de trigo (12,8%) e farinha de milho (14,3%). As determinações da urina revelaram a presença de AFM1 e AFP1, DON, OTA, FB1 e ZEN nos níveis de 0,02-12,0 ng/mg de creatinina. Em relação à ingestão provável diária (IPD) com base em dados de alimentos, apenas ZEN (0,156 µg/kg p.c./dia) apresentou um Quociente de Risco (HQ) acima do tolerável (> 1). Os valores de IPD baseados nos níveis urinários para DON, OTA e AF total foram 84,914, 0,031 e 0,001 µg/kg p.c./dia, respectivamente, resultando em valores de HQ> 1, o que pode indicar riscos para a saúde da população estudada. Uma intervenção informal por meio de atividades educacionais e entrega de um folheto informativo durante o primeiro período de amostragem não alterou os níveis de exposição às micotoxinas no segundo período de amostragem. Mais estudos são necessários para identificar itens alimentares além dos analisados neste trabalho como fontes de micotoxinas diárias, bem como a contribuição da inalação de pós contaminados para a exposição. Este é o primeiro estudo a relatar a avaliação de risco de micotoxinas com base em alimentos e níveis urinários em áreas rurais no Brasil.

Due to the dangerous toxic effects of mycotoxin ingestion via food products, several studies were performed in order to estimate human exposure to dietary mycotoxins as important factor in risk ass...

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.

In vitro tests were performed to determine the ability of beer fermentation residue (BFR) containing Saccharomyces cerevisiae to bind aflatoxin B1 (AFB1), zearalenone (ZEA), ochratoxin A (OTA) and deoxynivalenol (DON). BFR was obtained from a microbrewery, dried and ground, resulting in approximately 1.0 × 1010 S. cerevisiae cells g− 1 BFR. Binding assays consisted of suspending BFR (100 mg) in 10 mL of buffer solution (pH 3.0 or 6.5) spiked with AFB1, ZEA, OTA or DON (2.0 μg mL− 1 of each mycotoxin), incubation (60 min, 25 °C) followed by centrifugation. The supernatants were analyzed by high performance liquid chromatography. BFR had higher binding capacity for ZEA (75.1% and 77.5% at pH 3.0 and 6.5, respectively), when compared with AFB1, OTA and DON (less than 60% and 40% at pH 3.0 and 6.5, respectively). BFR also produced linear isotherms for ZEA at both pH values, hence indicating a potential application of industrial fermentation by-products containing yeast cells in reducing the bioavailability of ZEA in contaminated feedstuffs. However, in vivo studies are required to prove its efficacy in livestock and poultry.

This study aimed to determine the aflatoxin B1 (AFB1) binding capacity of a beer fermentation residue (BFR) containing Saccharomyces cerevisiae cells, and the efficacy of BFR to ameliorate the toxic effects of AFB1 on performance, serum biochemistry, and histology of broilers. The BFR was collected from a microbrewery, and the yeast cells were counted, dried, and milled before it was used in the study. In vitro evaluation of the BFR was conducted using different concentrations of AFB1 (2.0, 4.0, 8.0, 16.0, and 32.0 μg AFB1/mL) and 100 mg/10 mL of BFR at pH 3.0 or 6.0. Two hundred 1-day-old male broilers (Ross 308) were assigned to chick batteries and allowed ad libitum access to feed and water. A completely randomized design was used with 5 replicate pens of 5 chicks assigned to each of 4 dietary treatments from hatch to 21 d, which included: 1) basal diet (BD), with no BFR or AFB1; 2) BD supplemented with 1% BFR; 3) BD supplemented with 2 mg AFB1/kg of feed; and 4) BD supplemented with 2 mg AFB1/kg feed and 1% BFR. Performance variables were determined weekly, while serum analyses were performed on d 14 and 21. At the end of the study, chicks were anesthetized with carbon dioxide, euthanized by cervical dislocation, and the kidney, liver, and bursa of Fabricius were removed for determination of relative weights, and for histological evaluation. In vitro assays showed that the higher the initial AFB1 concentration in solution, the greater the AFB1 amount adsorbed by BFR at both pHs tested. Feed intake, BW gain, and concentrations of albumin, total protein, and globulin increased (P < 0.05) in broilers fed BFR+AFB1 (Diet 4), when compared to the birds receiving only AFB1 (Diet 2). Although BFR was not able to reduce or prevent the effects of AFB1 on relative weights of kidneys and liver, it reduced the severity of histological changes in the liver and kidney caused by AFB1.

Aflatoxin B1 (AFB1) can cause carcinogenic, mutagenic, teratogenic and immunosuppressive effects in humans and animals. Several lactic acid bacteria species have the ability to bind AFB1 in vitro, showing a potential application for reducing the bioavailability of AFB1 in contaminated products. Thus, the aim of this study was to evaluate the capacity of Lactobacillus rhamnosus, non-viable and dried, in removing the AFB1 from a contaminated medium. L. rhamnosus were cultured in MRS broth, sterilized (121 ºC, 15 min.) to inactivate their metabolism and then dried by spray-drying or freeze-drying (lyophilization). Binding assays using AFB1 (1.0 µg/ml) and L. rhamnosus cells (1×1010 cells, in suspension or spray-dried or freeze-dried) were conducted at pH 3.0 and 6.0, room temperature and contact time of 60 min. Quantification of AFB1 was achieved by high performance liquid chromatography. Scanning electron microscope was also performed in order to analyze the drying effect on the atomized and lyophilized L. rhamnosus cells. For pH 3.0 and 6.0, there were no significant differences between AFB1 binding efficiency by L. rhamnosus cells in solution (45.9 ± 8.8% and 35.8 ± 7.7%, respectively) or freeze-dried (36.6 ± 7.1% and 27.2 ± 4.0%, respectively). However, the spray-dried cells lost completely the AFB1 binding capacity during atomization, which damaged the structural and functional properties of the bacterial cell wall. In conclusion, L. rhamnosus retained its AFB1 binding ability only when its cell wall remained intact as observed in the lyophilization procedure. Lyophilized L. rhamnosus cells therefore can be a practicable alternative for decontamination of food products susceptible to aflatoxin contamination.