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Aflatoxins are fungal metabolites found in feeds and foods. When the ruminants eat feedstuffs containing Aflatoxin B1 (AFB1), this toxin is metabolized and Aflatoxin M1 (AFM1) is excreted in milk. International Agency for Research on Cancer (IARC) classified AFB1 and AFM1 as human carcinogens belonging to Group 1 and Group 2B, respectively, with the formation of DNA adducts. In the last years, some epidemiological studies were conducted on cancer patients aimed to evaluate the effects of AFB1 and AFM1 exposure on cancer cells in order to verify the correlation between toxin exposure and cancer cell proliferation and invasion. In this review, we summarize the activation pathways of AFB1 and AFM1 and the data already reported in literature about their correlation with cancer development and progression. Moreover, considering that few data are still reported about what genes/proteins/miRNAs can be used as damage markers due to AFB1 and AFM1 exposure, we performed a bioinformatic analysis based on interaction network and miRNA predictions to identify a panel of genes/proteins/miRNAs that can be used as targets in further studies for evaluating the effects of the damages induced by AFB1 and AFM1 and their capacity to induce cancer initiation.

The conversion of aflatoxin B1 in feed consumed by cows into aflatoxin M1 in their milk poses a challenge to food safety, with milk being a popular staple food and because of the harmful effects of these compounds. This study aimed at reviewing scientific information about the extent of carry-over of AFB1 from feed to milk. A range of studies reported about correlations of carry-over with different factors, particularly with milk yield and AFB1 intake. The extent of carry-over considerably varies, being 1–2% on average, which may be as high as 6% in the case of increased milk production. Specific factors influencing transfer rates, including milk yield, somatic cell counts, aflatoxin B1 intake, source of contamination, seasonal effects, particle size of feed, and the effects of certain interventions, namely vaccination and the use of adsorbents, are identified as the most important and are discussed in this review. The different mathematical formulas describing carry-over and instances of their application are reviewed as well. These carry-over equations may lead to largely different results, and no single carry-over equation can be suggested as the best one. While the exact quantification of carry-over is difficult as the process depends on many factors, including individual variabilities between animals, the intake of aflatoxin B1 and milk yield seem to be the most important factors influencing the excreted amount of aflatoxin M1 and the rate of carry-over.

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.

This study presents a novel electrochemical aptasensor for ultrasensitive detection of aflatoxin M 1 (AFM 1 ) in milk samples, utilizing a gold electrode modified with chitosan-functionalized multiwalled carbon nanotube/graphene nanocomposite (CS/f-MWCNTs-Gr). The platform was fabricated through covalent immobilization of an amino-modified aptamer onto the nanocomposite surface, enhancing electronic transmission and biorecognition efficiency. Cyclic voltammetry (CV) confirmed stepwise electrode modification, while square wave voltammetry (SWV) quantified AFM 1 via suppression of the [Fe(CN) 6 ] 3−/4− redox signal upon target binding. Under optimized conditions, the sensor demonstrated a wide linear range (1–1000 nM) covering EU/US regulatory limits, an ultra-low detection limit (0.03 nM, 9.8 ng/L, below EU regulatory limit of 25 ng/kg), and exceptional specificity (> 90% signal suppression against interferents at 10-fold higher concentrations). The aptasensor exhibited high reproducibility (RSD = 5.4%, n  = 5) and retained 92% signal after 14-day storage. Validated in commercial milk, it achieved 96–106% recoveries with RSD < 4.9% ( n  = 5), outperforming reference methods in precision and practicality. This cost-effective platform shows significant potential for on-site monitoring of mycotoxins in dairy products.

The existence of aflatoxin M1 (AFM1) in raw milk results in economic losses and public health risks. This research aims to examine the capability of bentonite to adsorb and/or eliminate AFM1 from various raw milk types. In addition, the effects of numerous bentonites (HAFR 1, 2, 3 and 4) on the nutritional characteristics of the milk were studied. Our findings revealed that goat milk had the highest value of AFM1 (490.30 ng/L) in comparison to other milks. AFM1 adsorption was influenced by applying bentonite (0.5 and 1 g) in a concentration-dependent manner for different time intervals (from 0 to 12 h). The percentage of AFM1 reached the maximum adsorption level after 12 h to 100, 98.5 and 98% for bentonites HAFR 3, 1 and 2, respectively. HAFR 3 (1 g bentonite) presented higher adsorption efficiency than other bentonites used in the phosphate buffer saline (PBS) and milk. Residual levels of AFM1 reached their lowest values of 0 and 1.5 ng/L while using HAFR 3 in PBS and milk, respectively. With regard to the influence of bentonite on the nutritional characteristics of milk, there was an increase in fat, protein and solid non-fat ratio while using HAFR 3 and 4, yet decreased lactose in comparison with the control. Scanning Electron Microscopy and Fourier Transform-Infrared Spectroscopy both identified bentonites as superior AFM1 binders. The results demonstrated that bentonite, particularly HAFR 3, was the most effective adsorbent and could thus be a promising candidate for the decontamination of AFM1 in milk.

Mycotoxins have been linked to adverse health impacts, including liver cancer and kidney diseases. The objectives of the current study were to evaluate the dietary exposure of Lebanese adults to multi-mycotoxins (aflatoxin B1 (AFB1), aflatoxin M1 (AFM1), ochratoxin A (OTA), ochratoxin B (OTB), deoxynivalenol (DON), T-2 and HT-2) and to assess their associated health risks. Hence, a nationally representative sample of 449 participants aged 18-64 years old were interviewed to obtain their socio-demographic characteristics, food consumption data and exposure estimates. A food frequency questionnaire and 24 h-recall were used to collect data. The concentration of mycotoxins in all foods consumed by the participants was collected from previous national published studies. The estimated daily intake (EDI), the hazard quotient (HQ) and the margin of exposure (MOE) were calculated. The total exposure to AFB1, AFM1, OTA and DON was 1.26, 0.39, 4.10 and 411.18 ng/kg bw/day, respectively. The MOE to AFB1, AFM1, OTA and DON in the Lebanese food basket was 316, 1454, 3539 and 510, respectively, indicating high health-related risks. Per food items, the MOE to AFB1 was below 10,000 in cereals (466.5), mainly in rice (827.9) and Burgul (4868.5). Similarly, the MOE to OTA in cereals was 1439, in which bread (4022), rice (7589) and bulgur (7628) were considered unsafe. Moreover, the MOE to DON in cereals (605) is alarming, especially in bread (632) and manakesh (6879). The MOE to AFM1 in dairy products was 1454, indicating health-related risks with a focus on yogurt (9788) and labneh (8153). As for the herbs/spices group and traditional dishes, the MOE to AFB1 was relatively lower than 10,000 (3690 and 1625, respectively), with a focus on thyme (2624) and kishik (3297), respectively. It is noteworthy that the MOE to DON and the MOE to OTA in traditional foods and coffee were lower than 10,000 (8047 and 8867, respectively). All hazard quotient (HQ) values were below 1, except the HQ value of milk and dairy products (1.96). The intake of some food groups varied between age categories, corresponding to differences in EDI between them. Thus, it is essential to put control measures in place to decrease the contamination and exposure to mycotoxins by Lebanese consumers.

This study explores the protective efficacy of Nigella sativa oil (NSO), Vitamin C, and E in countering the toxic effects of Aflatoxin M1 (AFM1) in neonatal Wistar rats. A total of 117 neonatal rats were randomized into 13 groups (A–M) and observed over 12 weeks. Group A served as the control, while the remaining groups were exposed to varying concentrations of AFM1, either alone or in combination with the supplements. Hematological parameters, including Packed Cell Volume (PCV), Red Blood Cells (RBC), Hemoglobin (HB), White Blood Cells (WBC), and liver enzyme activities, were measured using Microhematocrit Reader, Hemocytometer, Hemoglobinometer, and Spectrophotometer respectively. Histopathological assessments of liver tissues were conducted using staining techniques to assess cellular damage, inflammation, and fibrosis. The results demonstrated that simultaneous supplementation with NSO, Vitamin C, and E significantly improved PCV, HB, and RBC levels while reducing WBC counts and liver enzyme activities in comparison to AFM1-exposed groups. Notably, NSO effectively restored hematological parameters and facilitated liver tissue regeneration, even at higher concentrations of AFM1. In contrast, Vitamins C and E displayed protective effects predominantly at lower toxin levels. These findings suggest that NSO, in conjunction with Vitamin C and E, plays a pivotal role in mitigating the detrimental effects of AFM1 toxicity. NSO shows superior potential in reversing blood dyscrasia and liver damage. The study highlights the promising therapeutic utility of Nigella sativa oil as a natural remedy for alleviating aflatoxin-induced oxidative stress and organ damage, reinforcing its potential to safeguard public health against aflatoxin exposure.

A magnetic covalent organic framework (M-COF) was designed and selected as sorbent for magnetic solid-phase extraction (MSPE) of AFM1 and AFM2 in milk, followed by LC–MS/MS analysis. The application of 2,5-Dihydroxy-1,4-benzenedicarboxaldehyde (Dt) and 4′,5′-bis(4-aminophenyl)-[1,1′:2′,1″-terphenyl]-4,4″-diamine (BAPTPDA) as monomers endows M-COF excellent properties for adsorbing AFM1 and AFM2. The morphology, structure, stability, and magnetism of the Fe 3 O 4 @COF(BAPTPDA-Dt) were characterized by various techniques including scanning electron microscopy, transmission electron microscopy, FTIR, thermogravimetric analysis, and vibrating sample magnetometer. The Fe 3 O 4 microspheres were covered by COF shells. Fe 3 O 4 @COF exhibited excellent magnetism and stability. Some parameters that may influence the adsorption efficiency of MSPE were also optimized, making the extraction process more effective, time-saving (about 3 min), and less organic-reagent-consuming (only 4 mL of acetonitrile required). It is noteworthy that the Fe 3 O 4 @COF(BAPTPDA-Dt) can be reutilized more than 8 times. The AFM1 and AFM2 were determined by LC–MS/MS. The LODs for AFM1 and AFM2 were in the range 0.0069 to 0.0078 μg kg −1 . A wide linearity range (0.01–100 μg kg −1 ) with coefficients of determination ( R 2 ) ranging from 0.9998 to 0.9999 was obtained. The recoveries at four spiked concentrations (0.05, 0.5, 5, and 50 μg kg −1 ) in the milk matrix ranged from 85.2 to 106.5%. The intraday RSDs and the interday RSDs were in the range 1.74–4.58% and 2.65–6.69%, respectively. The matrix effect (9.3% for AFM1 and 6.7% for AFM2) was also significantly lower than that observed in other work . Overall, the established method has provided a powerful tool for rapid pretreatment and sensitive determination of AFM1 and AFM2 in milk with negligible matrix effect, presenting important value in toxicant determination. Graphical abstract

Aflatoxins (AFs) are fungal metabolites that are found in feed and food. When ruminants eat feed contaminated with aflatoxin B1 (AFB1), it is metabolised and aflatoxin M1 (AFM1) is excreted in the milk. Aflatoxins can result in hepatotoxic, carcinogenic, and immunosuppressive effects. The European Union thus set a low threshold limit (50 ng/L) for presence of AFM1 in milk. This was in view of its possible presence also in dairy products and that quantification of these toxins is mandatory for milk suppliers. In the present study, a total of 95,882 samples of whole raw milk, collected in northern Italy between 2013 and 2021, were evaluated for presence of AFM1 using an ELISA (enzyme-linked immunosorbent assay) method. The study also evaluated the relationship between feed materials collected from the same farms in the same area during the same period (2013–2021) and milk contamination. Only 667 milk samples out of 95,882 samples analysed (0.7%) showed AFM1 values higher than the EU threshold limit of 50 ng/L. A total of 390 samples (0.4%) showed values between 40 and 50 ng/L, thus requiring corrective action despite not surpassing the regulatory threshold. Combining feed contamination and milk contamination data, some feedingstuffs seem to be more effective in defying potential carryover of AFs from feed to milk. Combining the results, it can be concluded that a robust monitoring system that covers both feed, with a special focus on high risk/sentinel matrices, and milk is essential to guarantee high quality and safety standards of dairy products.

The manifestation of aflatoxins in feed and food is a major issue in the world as its presence leads to some health problems. This study investigates the incidence of aflatoxin M1 (AFM1) contamination in raw milk samples which were collected from Punjab, Pakistan. The Cluster Random Sampling technique was used to collect 960 milk samples from five different regions, and samples were collected every month. The AFM1 level in raw milk was analyzed by the ELISA technique. The findings demonstrate that 70% of samples exceeded the United States permissible maximum residue limits (MRL 0.50 µg/L), with an overall AFM1 level that ranged from 0.3 to 1.0 µg/L. AFM1 contamination varied with the season: The highest average contamination was detected in winter (0.875 µg/L), followed by autumn (0.751 µg/L), spring (0.654 µg/L), and summer (0.455 µg/L). The Eastern region exhibited the highest average AFM1 contamination (0.705 µg/L). Milk samples from the Northern region were found to be widely contaminated, as 86.9% samples exceeded the US MRL, followed by the Eastern region, with 72.3% samples being contaminated with >0.5 µg/L AFM1. The study indicated that the raw milk supply chain was heavily contaminated. Recommendations and remedial measures need to be developed by regulatory authorities to improve the raw milk quality.