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Cheese is one of the most susceptible dairy foods to accumulating aflatoxins due to their high affinity to caseins. The consumption of cheese contaminated with high levels of aflatoxin M1 (AFM1) can be highly harmful to humans. The present work, based on high-performance liquid chromatography (HPLC), highlights the frequency and levels of AFM1 in coalho and mozzarella cheese samples (n = 28) from the main cheese-processing plants in Araripe Sertão and Agreste in the state of Pernambuco, Brazil. Of the evaluated cheeses, 14 samples were artisanal cheeses and the remaining 14 were industrial (manufactured) cheeses. All samples (100%) had detectable levels of AFM1, with concentrations ranging from 0.026 to 0.132 µg/kg. Higher levels (p < 0.05) of AFM1 were observed in artisanal mozzarella cheeses, but none of the cheese samples exceed the maximum permissible limits (MPLs) of 2.5 µg/kg established for AFM1 in cheese in Brazil and 0.25 µg/kg in the European countries by the European Union (EU). The high incidence of low levels of AFM1 found in the evaluated cheeses underscores the need for stringent control measures to prevent this mycotoxin in milk used for cheese production in the study area, with the aim of protecting public health and reducing significant economic losses for producers. [ABSTRACT FROM AUTHOR]

The aim of this study was to conduct an up-to-date investigation on the occurrence levels of aflatoxin M1 (AFM1) in samples of raw milk (n = 40), pasteurized milk (n = 44), ultra-high temperature (UHT) milk (n = 27), Minas cheese (n = 57), and yogurt (n = 44) traded in São Paulo state, Brazil. AFM1 was extracted from fluid milks and dairy products using immunoaffinity columns and determined by high performance liquid chromatography. AFM1 was detected at the mean level of 0.080 ± 0.071 µg/L or kg in 72 samples (34.0%) evaluated in the study (n = 212). Detectable levels of AFM1 were observed in five samples of raw milk (12.5%), 16 samples of pasteurized milk (36.4%), 13 samples of UHT milk (48.1%), 27 samples of cheese (47.4%), and 11 samples of yogurt (25.0%), although none of them had concentrations above the maximum permitted levels (MPL) for AFM1 adopted in Brazil. However, 11.7% (n = 13) of samples of raw, pasteurized, and UHT milks would have AFM1 concentrations above the MPL of 0.05 μg/L adopted in the EU. The maximum level was detected in one cheese sample containing 0.695 µg/kg. Although none of the samples exceeded the Brazilian MPL, the high frequencies of AFM1 in Brazilian milk products warrant concern about their contribution to the human exposure to aflatoxins. Because aflatoxins are among the most potent carcinogens known, the results of this trial stress the need for stringent measures in the milk production system to avoid AFM1 in milk and derived products. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the antimicrobial effects of crude and lyophilized extracts of leaves from sweet passion fruit (Passiflora alata), araçá (Psidium cattleianum), rosemary (Rosamrinus officinalis) and oregano (Origanum vulgare) on planktonic cells of Staphylococcus aureus and Aspergillus parasiticus. Sweet passion fruit showed no inhibitory effect against the micro-organisms tested. However, crude and lyophilized extracts from Araçá had the highest (P < 0.05) antimicrobial activity against S. aureus, with minimum inhibitory concentrations (MIC) of 0.39 and 0.35 mg/ml, respectively. MIC values against S. aureus for lyophilized extracts from rosemary and crude extracts from oregano were 0.57 and 0.65 mg/ ml, respectively. None of the extracts demonstrated effective results against A. parasiticus, although araçá and oregano extracts had the lowest (P < 0.05) MIC values when compared with the other extracts. This preliminary screening study indicated that araçá, rosemary and oregano are interesting alternatives as antimicrobial agents in food substrates, although further studies are needed to develop commercial formulations based on field trials. [ABSTRACT FROM AUTHOR]

This work reports the chemical synthesis of aflatoxin B1 (AFB1)-lysine based on procedures available in the literature, but using lysine without a protection group in the α-amine group. AFB1-exo-8,9-epoxide was obtained by epoxidation of AFB1 with chloroperoxybenzoic acid in dichloromethane and phosphate buffer. Purification and identification of the AFB1-lysine were conducted by liquid chromatography (LC), and its structure was confirmed by LC with mass spectrometer and diode-array detection. The preparation of AFB1-lysine using lysine without a protection group in the α-amine group was completed in 24 h, being a practical modification of available methods that can be reproduced in analytical laboratories. [ABSTRACT FROM AUTHOR]

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 M 1 (AFM 1 ) in milk and aflatoxin B 1 (AFB 1 ), 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 AFB 1 by A. parasiticus was inhibited when the fungus was inoculated simultaneously with Lactiplantibacillus plantarum 3QB361 or L. plantarum 3QB350. No AFB 1 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 AFB 1 , 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 AFM 1 , 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., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Møller, Freire, Rosim, Margalho, Balthazar, Franco, Sant’Ana, Corassin, Rattray and Oliveira.)

The purpose of this study was to evaluate the ability of heat-killed cells (121 °C, 10 min) from two strains of lactic acid bacteria (LAB) (Lactobacillus rhamnosus and Lactococcus lactis) and one strain of yeast (Saccharomyces cerevisiae), alone or in combination, to reduce the levels of aflatoxin M 1 (AFM 1 ) in Frescal cheese during 30 days of storage. The experimental design was totally randomized, in a 2 × 2 × 2 factorial arrangement, corresponding to two levels of LAB (0 and L. rhamnosus at 10 10 cells/kg + L. lactis at 10 10 cells/kg), two levels of S. cerevisiae in milk (0 and 10 10 yeast cells/kg) and two AFM 1 levels (0 and 0.5 µg/kg) added to the cheese curd, totaling 8 treatments with three replicates per treatment. AFM 1 levels in Frescal cheese were evaluated by using a high-performance liquid chromatography. Cheese fat and protein contents were not affected (P > 0.05) by any of the treatments, and only pH decreased (P < 0.05) in all treatments from days 2 to 30 of storage (usual shelf life of this type of cheese). AFM 1 levels detected in contaminated cheeses decreased on day 2 of storage, varying from 0.09 µg/kg (cheese with addition of bacterial cells) to 0.29 µg/kg (no addition of LAB or yeast cells), this may have occurred due to loss of AFM 1 in the Frescal cheese whey. The concentrations of detected AFM 1 decreased (P < 0.05) in all treatments from days 2 to 10 of storage, and the maximum percentage reduction of the detectable levels (100%) was achieved after 10 and 20 days of storage in cheeses containing LAB and yeast cells, or prepared with yeast cells alone, respectively. The addition of heat-killed LAB (cells of L. rhamnosus and L. lactis) and Saccharomyces cerevisiae alone or in combination, has a potential ability for adsorbing the AFM 1 in Frescal cheese during 30 days of storage., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)