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Aflatoxins are carcinogens that can contaminate food and affect various body organs especially liver and kidney. When consumed, aflatoxin B 1 (AFB 1 ) is partially metabolised into aflatoxin M 1 (AFM 1 ), which is excreted in the urine. Breast milk may also contain AFM 1 due to maternal dietary intake from contaminated food. This cross-sectional study aimed to determine the levels of AFM 1 in both urine and breast milk among breastfeeding mothers ( n  = 256). The mother's demographic information was collected during recruitment. Mothers were then scheduled for an appointment to provide a morning urine sample along with five to ten mL samples of breast milk. AFM 1 levels in both samples were analysed using an enzyme-linked immunosorbent assay (ELISA). Spearman's rho and Chi-square were used to determine the associations between mean levels of AFM 1 in urine and breast milk. Findings show 68.0% of urine samples were contaminated with AFM1 (mean levels = 0.08 ± 0.04 ng/mL), while 14.8% of breast milk samples had AFM 1 (mean levels = 5.94 ± 1.81 ng/kg). Urine AFM 1 levels were not significantly associated with AFM 1 levels in breast milk ( p  > 0.05). This study can act as a baseline for future research examining long-term aflatoxin exposure among both mothers and infants.

Milk is an essential part of the human diet and is a nutrient-rich food that improves nutrition and food security. The aim of this study was to determine the presence and concentration of aflatoxin M1 (AFM1), adulterants, microbial loads, and physicochemical properties of raw cow's milk (CM) in Nekemte City, Ethiopia. A total of 12 samples of fresh CM were purposefully collected from four kebeles in the city (Bake Jama, Burka Jato, Cheleleki, and Bakanisa Kese) based on the potential of each milk production and distributor site. The AFM1 concentration was determined by high-performance liquid chromatography (HPLC) with a Sigma-Aldrich standard (St. Louis, MO, USA). The concentrations of AFM1 in Bake Jama, Burka Jato, Cheleleki, and Bakanisa Kese were found to be 0.01-0.03 g/L, 0.31-0.35 g/L, 0.19-0.21 g/L, and 0.04-0.07 g/L, respectively. The concentrations of AFM1 in the present study varied significantly ( p < 0.05) and ranged from 0.01 g/L to 0.35 g/L. These results show that of the 12 samples tested, all were positive for AFM1 and contaminated to varying degrees. The results of this study also revealed that the concentration of AFM1 in 7 (58%) of the 12 milk samples was above the European Union's (EU) maximum tolerance limit (0.05 g/L). The present study also revealed that of the investigated adulterants, only the addition of water had positive effects on three milk samples, while the remaining adulterants were not detected in any of the milk samples. The total bacterial count (TBC) and total coliform count (TCC) were significantly ( p < 0.05) different and ranged from 5.53 to 6.82 log 10 cfumL -1 and from 4.21 to 4.74 log 10 cfumL -1 , respectively. The physicochemical properties of the milk samples in the present study were significantly ( p < 0.05) different and ranged from 2.8% to 5.75% fat, 7.03% to 9.75% solid-not-fat (SNF), 2.35% to 3.61% protein, 3.33% to 5.15% lactose, 11.54% to 13.69% total solid, 0.16% to 0.18% titratable acid, 26.7 to 32.1°C, 6.35 to 6.55 pH, and 1.027 to 1.030 specific gravity. The physicochemical parameters of the raw milk in the study area met the required quality standards. Hence, further studies are required to determine the extent of the problem and the factors associated with high levels of AFM1 in raw milk in the study areas, including the detection of aflatoxin B1 (AFB1) in animal feed., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2024 Leila Nasir and Habtamu Fekadu Gemede.)

Objective: To understand the contents of aflatoxins(AFs) in foods sold in Shanghai, and to assess the exposure assessment of and its potential health risk among residents over 15 years old in Shanghai., Methods: A total of 8114 samples from 8 categories of food were collected in Shanghai from 2018 to 2023. The samples were detected by GB 5009.24-2016 and GB 5009.22-2016. Combined with the food consumption data of "Shanghai Diet and Health Survey", the dietary exposure assessment of aflatoxin was conducted using the margin of exposure(MOE) and the risk of liver cancer., Results: The detection rates of aflatoxin B&#95;1(AFB&#95;1), aflatoxin B&#95;2(AFB&#95;2), aflatoxin G&#95;1(AFG&#95;1), aflatoxin G&#95;2(AFG&#95;2), and aflatoxin M&#95;(1 )(AFM&#95;1) were 8.6%, 2.0%, 0.9%, 0.2% and 0.2%, respectively. The point assessment showed that the total exposure of AFB&#95;1 in the diet of residents aged 15 and above in Shanghai was 0.783 ng/(kg·BW·d), with the contribution rates of dietary exposure to grains and their products, nuts and their products, and vegetable oils accounting for 60.6%, 25.0% and 8.5% of AFB&#95;1's dietary exposure, respectively. The total exposure of total aflatoxins(the sum of AFB&#95;1, AFB&#95;2, AFG&#95;1 and AFG&#95;2)(AFT) was 7.363 ng/(kg·BW·d), and the dietary exposure of grains and their products, vegetable oils, nuts and their products contribute 77.1%, 8.4% and 7.2% to the dietary exposure of AFT, respectively. The probability assessment result indicated that the average dietary exposure of residents to AFB&#95;1 and AFT were 0.734 and 7.220 ng/(kg·BW·d), respectively, and the P95 exposure of residents were 1.170 and 11.500 ng/(kg·BW·d). The AFB&#95;1 exposure level of residents in suburban areas was higher than that in central urban areas and exurban areas(χ~2= 16.357, P&lt;0.001), the AFT exposure of residents in the central urban area was lower than that in the exurban areas and suburban areas(χ~2= 40.996, P&lt;0.001). According to the MOE method, the MOE values of AFB&#95;1 and AFT average dietary exposure for residents aged 15 and above in Shanghai were 511 and 54. The risk of liver cancer caused by dietary exposure of AFB&#95;1 and AFT among residents aged 15 and above in Shanghai were 0.024 cases/10~5 people and 0.227 cases/10~5 people., Conclusion: There is AFs contamination in food sold in Shanghai, and grains and their products, nuts and their products, and vegetable oils are the main sources of AFs exposure in the diet of residents aged 15 and above in Shanghai.

This study aimed to determine 16 mineral elements (Cd, Pb, As, Na, Mg, Al, Ca, K, Cr, Mn, Fe, Ni, Cu, Zn and Se) using inductively coupled plasma mass spectrometry (ICP-MS) and a direct mercury analyzer (DMA-80) for Hg evaluation. Aflatoxin M1 was determined by high-performance liquid chromatography with fluorescence detection (HPLC-FLD) in cow milk samples. This research considered 180 milk samples, 20 by province (Palermo, Catania, Messina), collected for a period of three years (2020-2022) to assess the potential risks for consumer, the safety status and nutritional quality related to mineral intake by consuming of milk. All samples showed a Pb concentration below the limit reported by European Regulation 915/2023. Cadmium and Hg concentrations were below the Limit Of Quantification (LOQ) in all samples analyzed. The milk samples analyzed proved to be a good source of Ca (up to 44.5 % of the dietary reference values), with well percentages also for Na (up to 7.6 %), K (up to 23.1 %) and Mg (up to 11.1 %). Regarding trace elements, the results reported that chromium requires attention; its value was always higher than 168.8 % in all samples analyzed. Levels of arsenic and lead were up to 20.2 % and up 7.1 % respectively. Aflatoxin M1 concentrations were below the limit of detection (< 0,009 mcg/kg) in all milk analyzed. Therefore, further studies are needed to safeguard consumer health, the quality of the product and to assess the state of animal health., Competing Interests: Declaration of Competing Interest All authors declare that there are no conflicts of interest in the development, drafting and publication of the following manuscript., (Copyright © 2024. Published by Elsevier GmbH.)

Co-contamination of multiple mycotoxins produces synergistic toxic effects, leading to more serious hazards. Therefore, the simple, rapid and accurate simultaneous detection of multiple mycotoxins is crucial. Herein, a three-channel aptamer-based lateral flow assay (Apt-LFA) was established for the detection of aflatoxin M 1 (AFM 1 ), aflatoxin B 1 (AFB 1 ) and ochratoxin A (OTA). The multi-channel Apt-LFA utilized gold‑iridium nanozyme to catalyze the chromogenic substrate, which effectively achieved signal amplification. Moreover, the positions and lengths of the complementary sequences were screened by changes in fluorescence intensity. After grayscale analysis, the semi-quantitative results showed that the detection limits of AFM 1 , AFB 1 and OTA were 0.39 ng/mL, 0.36 ng/mL and 0.82 ng/mL. The recoveries of the multiplexed competitive sensors in complex matrices of real samples were 93.33%-97.01%, 95.72%-102.67%, and 106.88%-109.33%, respectively. In conclusion, the assembly principle of the three-channel Apt-LFA is simple, which can provide a new idea for the simultaneous detection of small molecule targets., 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 © 2024. Published by Elsevier Ltd.)

Breast milk is one of the many distinct forms of food that can be contaminated with aflatoxin M 1 (AFM 1 ). They may be consumed by eating contaminated foods, such as contaminated meat and crops, which would then be present in breast milk and cause health problems, including nervous system disorders and cancers of the lungs, liver, kidneys, and urinary tract. However, the prevalently inconsistent explanation of prevalence and concentration remains a big challenge. Thus, this meta-analysis was conducted to determine the prevalence and concentration of harmful chemicals in breast milk in an African context. The databases MEDLINE, PubMed, Embase, SCOPUS, Web of Science, and Google Scholar were searched for both published and unpublished research. To conduct the analysis, the collected data were exported to Stata version 18. The results were shown using a forest plot and a prevalence with a 95% confidence interval (CI) using the random-effects model. The Cochrane chi-square (I2) statistics were used to measure the studies' heterogeneity, and Egger's intercept was used to measure publication bias. This review included twenty-eight studies with 4016 breast milk samples and newborns. The analysis showed the overall prevalence and concentration of aflatoxin M 1 in breast milk were 53% (95% CI 40, 65; i 2  = 98.26%; P = 0.001). The pooled mean aflatoxin M 1 concentration in breast milk was 93.02 ng/l. According to this study, the eastern region of Africa was 62% (95% CI 39-82) profoundly affected as compared to other regions of the continent. In subgroup analysis by publication year, the highest level of exposure to aflatoxins (68%; 95% CI 47-85) was observed among studies published from 2010 to 2019. This finding confirmed that more than half of lactating women's breast milk was contaminated with aflatoxin M 1 in Africa. The pooled mean aflatoxin M 1 concentration in breast milk was 93.02 ng/l. According to this study, the eastern region of Africa was profoundly affected compared with other regions. Thus, the government and all stakeholders must instigate policies that mitigate the toxicity of aflatoxins in lactating women, fetuses, and newborns., (© 2024. The Author(s).)

Given the highly mutagenic and carcinogenic nature of Aflatoxin M1 (AFM1), the quantity assessment of AFM1 residues in milk and dairy products is necessary to maintain consumer health and food safety. Herein, CRISPR-Cas12a-based colorimetric aptasensor was developed using the catalytic activity of flower-like nanozymes of MnO 2 and trans-cleavage property of CRISPR-Cas12a system to quantitatively detect AFM1. The basis of the developed colorimetric aptasensor relies on whether or not the CRISPR-Cas12a system is activated, as well as the contrast in oxidase-mimicking capability exhibited by flower-like MnO 2 nanozymes when AFM1 is absent or present. When AFM1 is not present in the sample, single-stranded DNA (ssDNA) is degraded by the activated CRISPR-Cas12a, and the solution turns into yellow due to the catalytic activity of the nanozymes. While, in the attendance of AFM1, ssDNA degradation does not occur due to the inactivation of the CRISPR-Cas12a. Therefore, with the adsorption of the ssDNA on the MnO 2 nanozymes, their catalytic activity decreases, and the solution color becomes pale yellow due to less oxidation of the chromogenic substrate. In this aptasensor, the relative absorbance changes increased linearly from 6 to 160 ng L -1 , and the detection limit was 2.1 ng L -1 . The developed aptasensor displays a selective detection performance and a practical application for quantitative analysis of AFM1 in milk samples. The results of the introduced aptasensor open up the way to design other selective and sensitive aptasensors for the detection of other mycotoxins by substitution of the used sequences., 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 © 2024 Elsevier B.V. All rights reserved.)

Aflatoxin M1 (AFM1) contamination of milk affects the general population with particular attention to children who frequently consume milk as part of complementary food. This study determined AFM1 contamination of cow's milk and estimated the health risk of dietary AFM1 through consumption of cow's milk among children (6 to 36 months) in the Magadu ward of Morogoro region in Tanzania. A total of 165 mother-baby pairs were recruited and interviewed on child feeding practices with a focus on feeding of cow's milk in the past 24 h. Alongside the interview, 100 raw cows' milk samples were collected from subsampled respondent households and were analyzed for AFM1 using enzyme-linked immunosorbent assay (ELISA). The results showed that about 35% of the surveyed children consumed cow's milk in the form of plain milk, incorporated in porridge and/or tea. The amount consumed varied from 62.5 to 500 mls with a median of 125 (125, 250) mls at a frequency of 1 to 2 times a day. All raw cows' milk (100%) samples (n = 100) were found contaminated with AFM1 at concentrations ranging from 0.052 to 9.310 µg/L and median of 2.076 µg/L (1.27, 2.48). All samples were contaminated by AFM1 at levels above the limits of 0.05 µg/L of raw milk set by the Tanzania Bureau of Standard and the European Union, while 97% exceeded 0.5 µg/L set by the US Food and Drug Administration. Exposure to AFM1 due to consumption of cow's milk ranged from 0.0024 to 0.077 µg/kg bw per day with a median of 0.019 (0.0016, 0.026) µg/kg bw per day, while the margin of exposure (MOE) ranged from 5.19 to 166.76 and median 20.68 (15.33, 25.40) implying high risk of public health concern. This study recommends that advocacy on consumption of cows' milk to combat undernutrition in children should consider a holistic approach that considers the milk's safety aspect., (© 2024. The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.)

Breast milk (BM) is considered as the best source of nutrition which could have prevention effects on various diseases in the first years of a child. Along with nutritive compounds, presence of contaminants such as mycotoxins in BM could be transmitted into neonate. The aim of this study was to determine the occurrence, levels, and factors associated with the presence of aflatoxin M 1 (AFM 1 ) and ocratoxin a (OTA) in BM samples of nursing mothers in rural centers of Yazd, Iran. The presence and average AFM 1 and OTA concentration in 72 BM samples was measured by competitive ELISA. The demographic and diet parameters of nursing mothers were collected by a questionnaire and were analyzed using SPSS 18 software. AFM 1 and OTA were detected in 63 (87.5%) and 47 (65.2%) samples with the mean concentration levels of 19.46 ± 13.26 ng/L (ranges from 5.1 to 53.9) and 200 ± 160 ng/L (ranges from 100 to 2460), respectively. Of these, 32 samples (50.7%) for AFM 1 and 23 samples (48.9%) for OTA had values exceeding the limit set by the European Union regulation for infant foods (25 ng/L for AFM 1 and 500 ng/L for OTA). It was also found that the risk of AFM 1 and OTA occurrence in BM increased significantly with the consumption of beans, bread, cereals, fruit juice and crackers, and cream, respectively. This study showed that the estimated daily intake for AFM 1 and OTA by 1 month of age infants was 2.7 and 28.5 ng/kg bw/day, respectively, while, as the age of the infant increased, the values were lower and close to 0.9 and 9.9 ng/kg bw/day for AFM1 and OTA in 12 months of age infants, respectively. The high occurrence and noticeable levels of AFM 1 and OTA detected in this study indicated that some infants receive undesirable exposures to AFM 1 and OTA with breast milk. Therefore, it is recommended that mothers are advised to avoid certain foods during pregnancy and breastfeeding that are likely sources of mycotoxins., (© 2024. The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.)

This study aimed to compare AFM 1 occurrence in different cheese types produced by organic and conventional systems; and to evaluate the risk of food exposure to AFM 1. A total of 176 commercial cheeses of 17 types were analyzed, 84 of organic and 92 of conventional production. Determination of AFM 1 was performed by high- performance liquid chromatography (HPLC), being detected in 30.5% of samples, with 4.8% of organic cheese samples presenting quantifiable AFM 1 values between 0.88 and 1.50 μg/kg. On the other hand, 4.3% of conventional cheese samples with values between 0.79 and 6.70 μg/kg. Two conventional cheese samples were above the limit of AFM 1 allowed for cheeses by the Brazilian legislation. No statistical difference were found between organic and conventional cheeses regarding the occurrence (p = 0.1780) and concentration of AFM 1 (p = 0.1810), according to the Chi-square and the T test, respectively. Estimated daily intake (EDI) and hazard index (HI) of dietary exposure to AFM 1 were 0.26 ng/kg/day and 1.28 ng/kg/day, respectively, for conventional cheese samples, and 0.09 ng/kg/day and 0.47 ng/kg/day for organic samples, with no statistical difference for EDI (p = 0.1729) and HI (p = 0.1802) between the two production systems. Comparison between several cheese types from conventional and organic systems indicated that AFM 1 is an obstacle to dairy production. Control and prevention of AFM 1 contamination, as well as detoxification methods in the final products, are necessary. In the case of organic products, additional research is needed in order to determine which control and detoxification methods should be allowed in this production system., 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 © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

A novel ratiometric electrochemical aptasensor based on split aptamer and Au-reduced graphene oxide (Au-rGO) nanomaterials was proposed to detect aflatoxin M1 (AFM1). In this work, Au-rGO nanomaterials were coated on the electrode through the electrodeposition method to increase the aptamer enrichment. We split the aptamer of AFM1 into 2 sequences (S1 and S2), where S1 was immobilized on the electrode due to the Au-S bond, and S2 was tagged with methylene blue (MB) and acted as a response signal. A complementary strand to S1 (CS1) labeled with ferrocene (Fc) was introduced as another reporter. In the presence of AFM1, CS1 was released from the electrode surface due to the formation of the S1-AFM1-S2 complex, leading to a decrease in Fc and an increase in the MB signal. The developed ratiometric aptasensor exhibited a linear range of 0.03 μg L -1 to 2.00 μg L -1 , with a detection limit of 0.015 μg L -1 for AFM1 detection. The ratiometric aptasensor also showed a linear relationship from 0.2 μg L -1 to 1.00 μg L -1 , with a detection limit of 0.05 μg L -1 in natural milk after sample pretreatment, indicating the successful application of the developed ratiometric aptasensor. Our proposed strategy provides a new way to construct aptasensors with high sensitivity and selectivity., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Introduction: In Sidama, Ethiopia, animal-source foods can be difficult to access. Milk has important nutrients for child growth, but carries the risk of aflatoxin M 1 (AFM 1 ) contamination. AFM 1 is a metabolite of the mycotoxin aflatoxin B 1 (AFB 1 ) in dairy feed; cows secrete AFM 1 in milk when their feed contains AFB 1 produced by Aspergillus fungi in maize, nuts and oilseeds. It is unknown whether AFM 1 compromises child growth and health., Methods and Analysis: This protocol paper describes our study in Sidama to determine the impact of milk consumption and AFM 1 on child growth in the first 18 months of life. We will collect baseline and end-line data on dairy production, socioeconomic and nutritional factors of 1000 dairy-owning households with children ages 6-18 months at baseline; and gather samples of milk and dairy feed and child anthropometrics. We will conduct phone interviews every 6 months to ascertain changes in practices or child health. Dairy feed will be tested for AFB 1 ; milk for AFM 1 , pathogens and nutrients. Controlling for herd size, socioeconomic, nutritional and behavioural factors, we will determine the association between child anthropometrics and milk consumption, as well as AFM 1 exposure. We will examine whether AFM 1 exposure affects child growth in the first 18 months of life, and weigh the benefits and risks of milk consumption., Ethics and Dissemination: The protocol is approved by the Institutional Review Boards of the Ethiopian Public Health Institute (EPHI-IRB-481-2022), Michigan State University (STUDY00007996) and International Food Policy Research Institute (DSGD-23-0102). Written informed consent will be obtained from all participants, who may withdraw from the study at any time. Confidentiality of collected data will be given high priority during each stage of data handling. The study's findings will be disseminated through stakeholder workshops, local and international conferences, journal articles and technical reports., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY. Published by BMJ.)

Background: The assessment of risks related to food safety is becoming a challenge in developing countries with its consequent health hazards. Chemical risk assessment in dairy products is important to maintain consumer health locally and internationally. Since milk and dairy products are essential foods for a wide range of customers, mostly children, patients, and pregnant women, it is very important to estimate the risks of some chemical residues, such as pesticides, some heavy metals, and aflatoxins., Aim: This work aims to determine the levels of chemical contamination in milk and traditional Egyptian cheese., Methods: Heavy metals were determined in samples by atomic absorption spectrometry. GC-mass spectrometry (MS)/MS and LC-MS/MS were also used for measuring pesticide residues. The Aflatoxin M1 was determined by enzyme-linked immune-sorbent assay., Results: Raw milk samples were tested and showed elevated concentrations of lead and cadmium, (46% and 4%, respectively). The heavy metals detected in the Egyptian cheese samples were variable depending on the type of cheese. Moreover, p.p.-DDE phenofose was present in 45% and 29% of raw milk and Ras cheese samples, respectively. For Aflatoxin M1, only 7% of milk samples and 2.9% of Ras cheese samples exceeded the acceptable limits., Conclusion: More surveying and risk assessment of chemical residues in milk and milk products are essential for controlling health risks to consumers., Competing Interests: The authors declare that there is no conflict of interest.

The objectives of this study were to evaluate the exposure to a diet naturally contaminated with mycotoxins on lactation performance, animal health, and the ability to sequester agents (SA) to reduce the human exposure to AFM 1 . Sixty healthy lactating Holstein cows were randomly assigned to two groups: naturally contaminated diet without and with the addition of a SA (20 g/cow/d AntitoxCooPil® -60% zeolite-40% cell wall-). Each cow was monitored throughout lactation. The concentration of aflatoxin B 1 (AFB 1 ) in feed and M 1 (AFM 1 ) in milk, health status, and productive and reproductive parameters were measured. AFB 1 concentration in feed was very low (2.31 μg/kgDM). The addition of SA reduced the milk AFM 1 concentrations (0.016 vs. 0.008 μg/kg) and transfer rates (2.19 vs. 0.77%). No differences were observed in health status, production and reproduction performance. The inclusion of SA in the diet of dairy cows reduce the risk in the most susceptible population., 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 © 2024 Elsevier B.V. All rights reserved.)

Realizing the simultaneous speedy detection of multiple mycotoxins in contaminated food and feed is of great practical importance in the domain of food manufacturing and security. Herein, a fluorescent aptamer sensor based on self-assembled DNA double-crossover was developed and used for effective simultaneous quantitative detection of aflatoxins M 1 and B 1 by fluorescence resonance energy transfer (FRET). Fluorescent dye-modified aflatoxin M 1 and B 1 aptamers are selected as recognition elements and signal probes, and DNA double crosses are consistently locked by the aflatoxin aptamers, which results in a "turn-off" of the fluorescent signal. In the presence of AFM 1 and AFB 1 , the aptamer sequences are more inclined to form Apt-AFM 1 and Apt-AFB 1 complexes, and the fluorescent probes are released from the DNA double-crossing platform, leading to an enhanced fluorescent signal (Cy3: 568 nm; Cy5: 660 nm). Under the optimal conditions, the signal response of the constructed fluorescent aptamer sensor showed good linearity with the logarithm of AFM 1 and AFB 1 concentrations, with detection limits of 6.24 pg/mL and 9.0 pg/mL, and a wide linear range of 0.01-200 ng/mL and 0.01-150 ng/mL, respectively. In addition, the effect of potential interfering substances in real samples was analyzed, and the aptasensor presented a good interference immunity. Moreover, by modifying and designing aptamer probes, the sensor can be applied to high-throughput simultaneous screening of other analytes, providing a new approach for the development of fluorescent aptamer sensors., 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 © 2023. Published by Elsevier B.V.)

Aflatoxins are a class of highly toxic mycotoxins. Aflatoxin M 1 (AFM 1 ) is hydroxylated metabolite of aflatoxin B 1 , having comparable toxicity, which is more commonly found in milk. In this study, the whole genome sequencing of Bacillus pumilus E-1-1-1 isolated from feces of 38 kinds of animals, having aflatoxin M 1 degradation ability was conducted. Bacterial genome sequencing indicated that a total of 3445 sequences were finally annotated on 23 different cluster of orthologous groups (COG) categories. Then, the potential AFM 1 degradation proteins were verified by proteomics; the properties of these proteins were further explored, including protein molecular weight, hydrophobicity, secondary structure prediction, and three-dimensional structures. Bacterial genome sequencing combined with proteomics showed that eight genes were the most capable of degrading AFM 1 including three catalases, one superoxide dismutase, and four peroxidases to clone. These eight genes with AFM 1 degrading capacity were successfully expressed. These results indicated that AFM 1 can be degraded by Bacillus pumilus E-1-1-1 protein and the most degrading proteins were oxidoreductases., Competing Interests: Declaration of competing interest We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted., (Copyright © 2023. Published by Elsevier B.V.)

Aflatoxin M1 (AFM1) is a mycotoxin that is commonly found as a milk contaminant, and its presence in milk has been linked to cytotoxicity. The present study aimed to evaluate the acute cytotoxic effects of AFM1 on intestinal Caco-2 cells. Initially, we checked the morphology and viability of Caco-2 cells after treatment with different concentrations of AFM1 (5 ng/L, 50 ng/L, 250 ng/L, 500 ng/L, 1000 ng/L, and 2000 ng/L) for different time intervals (6 h, 12 h, and 24 h). It was found that AFM1 did not show any effect on cell morphology, but 10% decrease in viability above 1000 ng/L after 12 h. Furthermore, DCFDA assay showed increased ROS production after 6 h treatments. qPCR analysis showed an increased expression of epithelial-specific cytoskeleton marker genes, Cytokeratin, Villin, Vimentin, and JAM-1, and a decreased expression of tight junction protein genes, Claudin-1, Occludin, and ZO-1. Similarly, we found an increased expression of Cyp1a1 transcript with an increasing AFM1 concentration and incubation time. This gene expression analysis showed AFM1 can cause disruption of tight junctions between intestinal cells, which was further confirmed by a transwell experiment. In conclusion, consumption of AFM1-contaminated milk does not show any effect on cells morphology and viability but decreases the expression of intestinal barrier transcripts that may lead to the disruption of intestinal barrier function and leaky gut., (© 2023. The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.)

Mycotoxins are abiotic hazards whose contamination occurs at the pre- and post-harvest stages of the maize value chain, with animal exposure through contaminated feed leading to their excretion into milk. Currently, only aflatoxin M1 is regulated in milk products. Since feed materials and complete feed present a multi-mycotoxin composition and are the main mycotoxin source into milk, it is important to recognize the occurrence of multiple toxins and their co-occurrence in this highly consumed food product. The aim of this study was to determine the content of regulated and emerging mycotoxins in milk samples, which allowed for evaluating the occurrence and co-occurrence patterns of different mycotoxins known to contaminate feed materials and complete animal feed. Human exposure considering the occurrence patterns obtained was also estimated. Aflatoxins, fumonisins, zearalenone, and emerging mycotoxins were among the mycotoxins found to be present in the 100 samples analyzed. Concentrations ranged from 0.006 to 16.3 μg L -1 , with no sample exceeding the AFM1 maximum level. Though several mycotoxins were detected, no exceeding values were observed considering the TDI or PMTDI. It can be concluded that the observed exposure does not pose a health risk to milk consumers, though it is important to recognize vulnerable age groups.

The objective of this network meta-analysis was to determine the efficacy of different mycotoxin binders (MTB) to reduce aflatoxin M 1 (AFM 1 ) in milk. A literature search was conducted to identify in vivo research papers from different databases. Inclusion criteria were in vivo, dairy cows, description of the MTB used, doses of MTB, aflatoxin inclusion in the diet, and concentration of AFM 1 in milk. Twenty-eight papers with 131 data points were selected. Binders used in the studies were hydrated sodium calcium aluminosilicate (HSCAS), yeast cell wall (YCW), bentonite, and mixes of several MTB (MX). The response variables were AFM 1 concentration, AFM 1 reduction in milk, total AFM 1 excreted in milk, and transfer of aflatoxin from feed to AFM 1 in milk. Data were analyzed with CINeMA and GLIMMIX procedures with the WEIGHT statement of SAS (SAS Inst. Inc.). The AFM1 concentration in milk decreased for bentonite (0.3 µg/L ± 0.05; mean ± SE) and HSCAS (0.4 µg/L ± 0.12), and tended to decrease for MX (0.6 µg/L ± 0.13) but was similar for YCW (0.6 µg/L ± 0.12), compared with control (0.7 µg/L ± 0.12). The percentage reduction of AFM 1 in milk was similar for all MTB and different from control with a range of reduction from 25% for YCW to 40% for bentonite. The excretion of AFM 1 in milk was lower in YCW (5.3 µg/L ± 2.37), HSCAS (13.8 µg/L ± 3.31), and MX (17.1 µg/L ± 5.64), and not affected by bentonite (16.8 µg/L ± 3.33) compared with control (22.1 µg/L ± 5.33). The transfer of aflatoxin B 1 from feed into AFM 1 in milk was lowest in bentonite (0.6% ± 0.12), MX (1.04% ± 0.27), and HSCAS (1.04% ± 0.21), and not affected in YCW (1.4% ± 0.10), compared with control (1.7% ± 0.35). The meta-analysis results indicate that all MTB reduced the AFM 1 transfer into milk, where bentonite had the highest capacity and YCW the lowest., (© 2023, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Considering the genotoxic and cancerogenic nature of aflatoxin M1 (AFM1), its presence in milk and dairy products may pose health risks for consumers. The chronic exposure was calculated using a two-dimensional (second order) Monte Carlo model. Results of 13 722 milk and dairy product samples analysed in the 2015-2022 period were used. Milk and dairy products intake information was collected with a Food Frequency Questionnaire (FFQ) validated by a 24-h recall-based method. Risk characterization was done by calculation of the Margin of Exposure (MOE) and by calculation of AFM1 induced number of hepatocellular carcinoma (HCC) cases. Mean AFM1 Estimated Daily Intake (EDI) was highest in children at 0.336 (CI: 0.294-0.385) ng kg -1 bw day -1 , followed by adolescents with 0.183 (CI: 0.164-0.204), then adult females with 0.161 (CI: 0.146-0.179) and finally adult males with lowest EDI of 0.126 (CI: 0.115-0.139) ng kg -1 bw day -1 . MOE values based on mean EDI for all population groups were above risk associated threshold and the number of possible HCC cases was in the range of 0.0002-0.0021 cases per year for 10 5 individuals. The results suggest low health risks due to AFM1 exposure for the whole population. Still, this risk is not non-existent, especially for children as they have a higher ratio of the population exposed to risk associated AFM1 levels, with MOE values below risk indicating threshold starting at 77.5th percentile., 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 © 2023 Elsevier Ltd. All rights reserved.)

Aflatoxin M 1 (AFM 1 ) contamination poses a serious threat to human health globally. Hence, it is necessary to develop reliable and ultrasensitive methods for the determination of AFM 1 residue in food products at low levels. In this study, a novel polystyrene microsphere-mediated optical sensing (PSM-OS) strategy was constructed to solve the problems of low sensitivity and susceptibility to interference from the matrix in AFM 1 determination. Polystyrene (PS) microspheres have the advantages of low cost, high stability, and controllable particle size. They can be useful optical signal probes for qualitative and quantitative analyses attributed to the fact that they have strong ultraviolet-visible (UV-vis) characteristic absorption peaks. Briefly, magnetic nanoparticles were modified with the complex of bovine serum protein and AFM 1 (MNP 150 -BSA-AFM 1 ), and biotinylated antibodies of AFM 1 (AFM 1 -Ab-Bio). Meanwhile, PS microspheres were also functionalized with streptavidin (SA-PS 950 ). In the presence of AFM 1 , a competitive immune reaction was triggered leading to the changes in AFM 1 -Ab-Bio concentrations on the surface of MNP 150 -BSA-AFM 1 . The complex of MNP 150 -BSA-AFM 1 -Ab-Bio binds with SA-PS 950 to form the immune complexes due to the special binding of biotin and streptavidin. The remaining SA-PS 950 in the supernatant was determined by UV-Vis spectrophotometer after magnetic separation, which positively correlated with the concentration of AFM 1 . This strategy allows for ultrasensitive determination of AFM 1 with limits of detection as low as 3.2 pg/mL. It was also successfully validated for AFM 1 determination in milk samples, and a high consistency was found with the chemiluminescence immunoassay. Overall, the proposed PSM-OS strategy can be used for the rapid, ultrasensitive, and convenient determination of AFM 1 , as well as other biochemical analytes., 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 © 2023 Elsevier B.V. All rights reserved.)

Melamine (MEL), enrofloxacin (ENR), sulfamethazine (SMZ), tetracycline (TC), and aflatoxin M 1 (AFM 1 ) are the main chemical contaminants in milk. It is necessary to detect these miscellaneous chemical contaminants in milk synchronously to ensure the safety of the milk. In this study, a multiple lateral flow immunoassay (LFIA) was developed for the detection of MEL, ENR, SMZ, TC, and AFM 1 in milk. Under optimal experimental conditions, the cutoff values were 25 ng/mL for MEL, 1 ng/mL for ENR, 2.5 ng/mL for SMZ, 2.5 ng/mL for TC, and 0.25 ng/mL for AFM 1 in milk samples. The limits of detection of LFIA were 0.173 ng/mL for MEL, 0.078 ng/mL for ENR, 0.059 ng/mL for SMZ, 0.082 ng/mL for TC, and 0.0064 ng/mL for AFM 1 . The recovery rates of LFIA in milk were 83.2-104.4% for MEL, 76.5-127.3% for ENR, 96.8-113.5% for SMZ, 107.1-166.6% for TC, and 93.5-130.3% for AFM 1 . The coefficients of variation were all less than 15%. As a whole, the developed multiple lateral flow immunoassay showed potential as a highly reliable and excellent tool for the rapid and sensitive screening of MEL, ENR, SMZ, TC, and AFM 1 in milk., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

In this study, the average level of aflatoxin M1 in various types of milk from 107 articles (297 studies with 16,274 milk samples) were meta-analyzed using random-effect model based on the milk varieties (animal species and heating processes), geographical regions, seasons, detection techniques and dairy farming subgroups. Studies on milk contamination with aflatoxin M1 in Iran were collected using universal and Persian databanks from January 1974 to the end of November 2021. The overall aflatoxin M1 mean concentration and prevalence in milk samples of Iran were 39.65 ng/l (95% CI: 36.00-43.30) and 80% (95% CI: 76-85%), respectively. The rank order of importance of various variables in mean levels of aflatoxin M1 in milk samples included milk type (animal species) > geographical regions > detection techniques > dairy farming types > milk types (heating processes) > seasons. Findings revealed that the overall content of aflatoxin M1 in milk samples of Iran was lower than that allowed by the European Union, Institute of Standards and Industrial Research of Iran, and the USA, possibly due to the milk monitoring by the Iranian regulatory systems., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Aflatoxin M1 (AFM1) is detected in the milk of animals after ingestion of aflatoxin B1-contaminated food; since 2002, it has been categorized as a group I carcinogen. In this work, a silicon-based optoelectronic immunosensor for the detection of AFM1 in milk, chocolate milk, and yogurt has been developed. The immunosensor consists of ten Mach-Zehnder silicon nitride waveguide interferometers (MZIs) integrated on the same chip with the respective light sources, and an external spectrophotometer for transmission spectra collection. The sensing arm windows of MZIs are bio-functionalized after chip activation with aminosilane by spotting an AFM1 conjugate with bovine serum albumin. For AFM1 detection, a three-step competitive immunoassay is employed, including the primary reaction with a rabbit polyclonal anti-AFM1 antibody, followed by biotinylated donkey polyclonal anti-rabbit IgG antibody and streptavidin. The assay duration was 15 min with limits of detection of 0.005 ng/mL in both full-fat and chocolate milk, and 0.01 ng/mL in yogurt, which are lower than the maximum allowable concentration of 0.05 ng/mL set by the European Union. The assay is accurate (% recovery values 86.7-115) and repeatable (inter- and intra-assay variation coefficients <8%). The excellent analytical performance of the proposed immunosensor paves the way for accurate on-site AFM1 determination in milk.

Aflatoxin M 1 (AFM 1 ), a secondary metabolite of Aspergillus spp., is highly toxic and widely present in food matrices. Therefore, the detection of AFM 1 is of great importance for the protection of food safety. In this study, a five-segment sequence was designed as the initial library. Graphene oxide-SELEX (GO-SELEX) was applied to screen AFM 1 . After seven rounds of repeated screening, affinity and specificity assays showed that aptamer 9 was the best candidate for AFM 1 . The dissociation constant ( K d ) of aptamer 9 was 109.10 ± 6.02 nM. To verify the efficiency and sensitivity aptamer for the detection of AFM 1 , a colorimetric sensor based on the aptamer was constructed. The biosensor showed good linearity in the range of AFM 1 concentration of 0.5-500.0 ng/mL with a detection limit of 0.50 ng/mL. This colorimetric method was successfully used for the detection of AFM 1 in milk powder samples. Its detection recovery was 92.8-105.2%. This study was conducted to provide a reference for the detection of AFM 1 in food.

In this study, a chloramphenicol and aflatoxin M1 aptamer-functionalized DNA hydrogel was designed for the simultaneous detection of chloramphenicol and aflatoxin M1 for the first time. The acrydite-modified chloramphenicol aptamer sequence was used to synthesize the DNA hydrogel and for visual detection of chloramphenicol depending on the gel-to-sol transition of the target-responsive DNA hydrogel. The DNA hydrogel formulation was set as follows: 60% of each linear polyacrylamide-DNA conjugate and 40% of acrylamide and chloramphenicol aptamer/DNA strand-1 at a molar ratio of 1:1, and the lowest concentration of chloramphenicol leading to gel dissociation was 1.0 nM at 25 °C. Furthermore, the formalized aptamer-functionalized DNA hydrogel was used to detect aflatoxin M1 by measuring the recovery of the fluorescence signal that was quenched when the FAM-labeled aflatoxin M1 aptamer and BHQ1-labeled DNA strand-2 were hybridized to form a double-stranded DNA in the network of hydrogel. The detection platform was successfully applied to the detection of chloramphenicol and aflatoxin M1, both in aqueous solution and in milk. The aptamer-functionalized DNA hydrogel had detection (LOD) and quantification limits (LOQ) for aflatoxin M1 as 1.7 and 5.2 nM, respectively. Using two aptamer sequences with high affinity and specificity, the dual-sensing platform based on the DNA hydrogel achieved higher selectivity for chloramphenicol and aflatoxin M1, which demonstrated its potential as a reliable simultaneous detection platform against two different targets for monitoring food safety.

Background: Aflatoxin M1 (AFM1) is found in the milk of cows exposed to feed spoiled by Aspergillus fungi species. These fungi may produce the secondary metabolite aflatoxin B1, which is converted in the cow liver by hydroxylation to AFM1 and is then expressed in milk. AFM1 is regulated in milk and other dairy products because it can cause serious health issues, such as liver and kidney cancers, in humans and is an immunosuppressant., Objective: To optimize the chromatographic protocol and to extend the matrix scope to include a wider range of dairy products: whey powder, whey protein concentrate, whey protein isolate, liquid milk, skim milk powder, whole milk powder, adult nutritional products, and yogurt., Methods: AFM1 is extracted using 1% acetic acid in acetonitrile incorporating ionic salts. The AFM1 in the resulting extract is concentrated using an automated RIDA®CREST IMMUNOPREP® online cartridge coupled to quantification by HPLC-fluorescence., Results: The method was shown to be accurate, with acceptable recovery (81.2-97.1%) from spiked samples. Acceptable precision was confirmed, with a relative standard deviation (RSD) for repeatability of 6.6-11.2% and an RSD for intermediate precision of 7.5-16.7%. Method LOD and robustness experiments further demonstrated the suitability of this method for routine compliance testing. Analysis of an international proficiency trial sample generated results that were comparable with the value assigned from alternative independent methods., Conclusion: A method with improved chromatography for high-throughput, routine testing of AFM1 in an extended range of dairy products is described. The method was subjected to single-laboratory validation and was found to be accurate, precise, and fit for purpose., Highlights: Single-laboratory validation of an automated online immunoaffinity cleanup fluorescence HPLC method for AFM1 in whey proteins, milk powders, nutritional products, liquid milk, and yogurt. Allows for high-throughput analysis of AFM1 with enhanced chromatographic performance. Method applicable to the analysis of AFM1 in an extended range of milk and milk-based products., (© The Author(s) 2023. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

In the past few years there has been a growing trend in the prevalence of aflatoxins, attributable to climate change, in substances destined for animal feeding, together with an increase in dairy product consumption. These facts have triggered great concern in the scientific community over milk pollution by aflatoxin M 1 . Therefore, our study aimed to determine the transfer of aflatoxin B 1 from the diet into milk as AFM 1 in goats exposed to different concentrations of AFB 1 , and its possible effect on the production and serological parameters of this species. For this purpose, 18 goats in late lactation were divided into 3 groups (n = 6) and exposed to different daily doses of aflatoxin B 1 (T1 = 120 µg; T2 = 60 µg, and control = 0 µg), during 31 d. Pure aflatoxin B 1 was administered 6 h before each milking in an artificially contaminated pellet. The milk samples were taken individually in sequential samples. Milk yield and feed intake were recorded daily, and a blood sample was extracted on the last day of exposure. No aflatoxin M 1 was detected, either in the samples taken before the first administration, or in the control group ones. The aflatoxin M 1 concentration detected in the milk (T1 = 0.075 µg/kg; T2 = 0.035 µg/kg) increased significantly on a par with the amount of aflatoxin B 1 ingested. The amount of aflatoxin B 1 ingested did not have any influence on aflatoxin M 1 carryover (T1 = 0.066% and T2 = 0.060%), these being considerably lower than those described in dairy goats. Thus, we concluded that the concentration of aflatoxin M 1 in milk follows a linear relationship with respect to the aflatoxin B 1 ingested, and that the aflatoxin M 1 carryover was not affected by the administration of different aflatoxin B 1 doses. Similarly, no significant changes in the production parameters after chronic exposure to aflatoxin B 1 were observed, revealing a certain resistance of the goat to the possible effects of that aflatoxin., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

This study aimed to assess the prevalence, concentration of AFM1 in human breast milk, and to determine the risk of cancer for infants in sub-Saharan Africa. A systematic literature search was performed using PubMed, CINAHL, Web of science, global health, Cochrane, and Google Scholar electronic databases. A random-effects model was used to estimate the pooled prevalence and concentration of AFM1 in breast milk. The meta-analysis of 8 articles containing 9 studies showed the pooled prevalence of AFM1 in breast milk to be 56.18% (95% CI: 29.65-82.71) and the pooled concentration to be 31.12 ng/L (95% CI: 25.97-36.25). The cancer risk assessment indicated for both male and female 1-month infants in Sierra Leone (HI > 1) is high, and all the rest of the infants are free of risk (HI < 1). The pooled prevalence and mean concentration of AFM1 in breast milk is high. Monitoring of AFB1 concentration of commonly used foods will be of high value in reducing the burden of AFM1.

Objective: Mycotoxins are different toxic substances at relatively smaller molecular weight produced by some types of fungi. Aflatoxin is the most common type of mycotoxin easily reproducing in food stored for a long time in unsuitable conditions. This study determined the aflatoxin M1 (AFM1) levels in breast milk samples collected from mothers who gave birth in Kırşehir, Turkey., Materials and Methods: A total of 82 breast milk samples to be analyzed to determine the AFM1 levels were collected from voluntary breastfeeding mothers who gave birth in the Kırşehir Training and Research Hospital and who were randomly selected. The AFM1 levels were determined using the competitive ELISA kit., Results: The AFM1 levels in the breast milk samples of mothers who did not consume milk were lower than those of other mothers. The AFM1 levels in the breast milk samples of mothers consuming fabrication milk were lower than mothers consuming homemade milk (p<0.01). Additionally, the AFM1 levels in the breast milk samples of mothers consuming homemade or self-made bread were lower (p<0.05)., Conclusions: This study found that the nutritional habits of breastfeeding mothers affected the AFM1 levels in breast milk.

The aim of this work was to estimate the effects of three recombinant peroxidases (rPODs) on the degradation of aflatoxin M 1 (AFM 1 ) in a model solution and were applied in milk and beer to study the AFM 1 degradation. Besides, the contents of AFM 1 in model solution, milk and beer were evaluated, and the kinetic parameters of rPODs were determined (Michaelis-Menten constant - K m and maximal velocity - V max ). The optimized reaction conditions (The degradation was over 60 %) for these three rPODs in the model solution were, respectively as follows: pH were 9, 9, and 10; hydrogen peroxide concentrations were 60, 50, and 60 mmol/L; at an ionic strength of 75 mmol/L and reaction temperature of 30 °C with 1 mmol/L K + or 1 mmol/L Na + . These three rPODs (1 U /mL) presented the maximum activity for degradation of AFM 1 of 22.4 %, 25.6 %, and 24.3 % in milk, while 14.5 %, 16.9 %, and 18.2 % in beer, respectively. Meanwhile, the survival rate of Hep-G2 cells raised about 1.4 times after treated with peroxidase-generated AFM 1 degradation products. Therefore, POD may be a promising alternative to reduce the pollution of AFM 1 in model solution, milk, beer, and minimize their impact on the environment in humans., 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 © 2022 Elsevier Ltd. All rights reserved.)

Aflatoxin B1 (AFB1) and aflatoxin M1 (AFM1) are universally found as environmental pollutants. AFB1 and AFM1 are group 1 human carcinogens. Previous sufficient toxicological data show that they pose a health risk. The intestine is vital for resistance to foreign pollutants. The enterotoxic mechanisms of AFB1 and AFM1 have not been clarified at the metabolism levels. In the present study, cytotoxicity evaluations of AFB1 and AFM1 were conducted in NCM 460 cells by obtaining their half-maximal inhibitory concentration (IC50). The toxic effects of 2.5 μM AFB1 and AFM1 were determined by comprehensive metabolomics and lipidomics analyses on NCM460 cells. A combination of AFB1 and AFM1 induced more extensive metabolic disturbances in NCM460 cells than either aflatoxin alone. AFB1 exerted a greater effect in the combination group. Metabolomics pathway analysis showed that glycerophospholipid metabolism, fatty acid degradation, and propanoate metabolism were dominant pathways that were interfered with by AFB1, AFM1, and AFB1+AFM1. Those results suggest that attention should be paid to lipid metabolism after AFB1 and AFM1 exposure. Further, lipidomics was used to explore the fluctuation of AFB1 and AFM1 in lipid metabolism. The 34 specific lipids that were differentially induced by AFB1 were mainly attributed to 14 species, of which cardiolipin (CL) and triacylglycerol (TAG) accounted for 41%. AFM1 mainly affected CL and phosphatidylglycerol, approximately 70% based on 11 specific lipids, while 30 specific lipids were found in AFB1+AFM1, mainly reflected in TAG up to 77%. This research found for the first time that the lipid metabolism disorder caused by AFB1 and AFM1 was one of the main causes contributing to enterotoxicity, which could provide new insights into the toxic mechanisms of AFB1 and AFM1 in animals and humans.

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.

Monitoring aflatoxin M1 (AFM1) in dairy products and milk-based foods is very important. The main purpose of this research was to investigate and determine the amount and human health risk assessment of aflatoxin M1 (AFM1) in the most famous and widely used brands of infant dried powder milk (IDPM) consumed in Iran. For this study, 45 imported IDPM (IM-IDPM) samples and 45 domestically produced IDPM (DO-IDPM) samples (a total of 90 samples) were selected randomly. All samples were analyzed for AFM1 using a competitive enzyme-linked immunosorbent assay (ELISA) technique. The mean level and the percentage of positive samples for AFM1 in DO-IDPM were 9.2 ± 5.4 ng/kg and 73.3%, and for IM-IDPM, they were 5.1 ± 3.8 ng/kg and 33.3%, respectively. The average level of AFM1 in all samples was lower than the EU and Iranian national standards (25 ng/kg). AFM1 intake through IDPM consumption by Iranian infants less than one-year-old was lower than the allowable level, but the hazard quotient for infants less than 6 months was higher than the allowable level. Although the concentration of AFM1 in IDPM consumed in Iran is not hazardous, since Iranian infants may be exposed to AFM1 through other sources, including baby food, breast milk, raw and pasteurized milk, continuous monitoring of IDMP quality in Iranian markets should be taken., Competing Interests: Declaration of Competing Interests 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 © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

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 M 1 (AFM 1 ) can be highly harmful to humans. The present work, based on high-performance liquid chromatography (HPLC), highlights the frequency and levels of AFM 1 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 AFM 1 , with concentrations ranging from 0.026 to 0.132 µg/kg. Higher levels ( p < 0.05) of AFM 1 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 AFM 1 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 AFM 1 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.

Mycotoxins are compounds produced by several fungi that contaminate agricultural fields and, either directly or by carry-over, final food products. Animal exposure to these compounds through contaminated feed can lead to their excretion into milk, posing threats to public health. Currently, aflatoxin M1 is the sole mycotoxin with a maximum level set in milk by the European Union, as well as the most studied. Nonetheless, animal feed is known to be contaminated by several groups of mycotoxins with relevance from the food safety point of view that can be carried over into milk. To evaluate the multi-mycotoxin occurrence in this highly consumed food product it is crucial to develop precise and robust analytical methodologies towards their determination. In this sense, an analytical method for the simultaneous identification of 23 regulated, non-regulated, and emerging mycotoxins in raw bovine milk using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) was validated. A modified QuEChERS protocol for extraction purposes was used, and further validation was performed by assessing the selectivity and specificity, limits of detection and quantification (LOD and LOQ), linearity, repeatability, reproducibility, and recovery. The performance criteria were compliant with mycotoxin-specific and general European regulations for regulated, non-regulated, and emerging mycotoxins. The LOD and LOQ ranged between 0.001 and 9.88 ng mL -1 and 0.005 and 13.54 ng mL -1 , respectively. Recovery values were between 67.5 and 119.8%. The repeatability and reproducibility parameters were below 15 and 25%, respectively. The validated methodology was successfully applied to determine regulated, non-regulated, and emerging mycotoxins in raw bulk milk from Portuguese dairy farms, proving the importance of widening the monitoring scope of mycotoxins in dairy products. Additionality, this method presents itself as a new strategic and integrated biosafety control tool for dairy farms for the analysis of these natural and relevant human risks.

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 objective of the study was to investigate the efficacy of bentonite and fumonisin esterase, separately or combined, in mitigating the effects of aflatoxins (AF) and fumonisins (FUM) in Boran and Friesian-Boran crossbreed cattle. These effects were studied by measuring mycotoxins, their metabolites, and biomarkers that relate to animal health, productivity, and food safety. The study was divided into three experiments each lasting for 2 weeks. Cows in experiment 1 received in random order aflatoxin B1 (AFB1) [788 μg/cow/day (69.7 μg/kg dry matter intake (DMI)) for Borans and 2,310 μg/cow/day (154 μg/kg DMI) for crossbreeds], bentonite (60 g/cow/day), or both AFB1 and bentonite. Boran cows in experiment 2 received in random order FUM (12.4 mg/cow/day (1.1 mg/kg DMI)), fumonisin esterase (120 U/cow/day), or both FUM and fumonisin esterase. Boran cows in experiment 3 received in random order AFB1 (952 μg/cow/day (84.2 μg/kg DMI)) + FUM (30.4 mg/cow/day (2.7 mg/kg DMI)), bentonite (60 g/cow/day) + fumonisin esterase (120 U/cow/day), or both AFB1 + FUM and bentonite + fumonisin esterase. Feeding AFB1 and/or FUM contaminated feed with or without the addition of the detoxifiers for 14 days did not affect DMI, milk composition, hematology, and blood biochemical parameters. The addition of bentonite in a diet contaminated with AFB1 led to a decrease in milk aflatoxin M1 (AFM1) concentration of 30% and 43%, with the carry-over subsequently decreasing from 0.35% to 0.20% and 0.08% to 0.06% for crosses and Borans, respectively. No significant change was observed in the sphinganine/sphingosine (Sa/So) ratio following feeding with FUM alone or in combination with fumonisin esterase; however, the ability of fumonisin esterase to hydrolyze FUM into less toxic fully hydrolyzed FUM and partially hydrolyzed FUM was evident in the rumen fluid and feces. These results indicate bentonite was effective in decreasing AFM1 concentration in milk, and AFB1 and AFM1 in plasma, while fumonisin esterase can convert FUM into less toxic metabolites and can be a suitable addition to feed cocontaminated with AFB1 and FUM.

In the study, Cu 2-x Se-Au nanoparticles (CSA) with a photothermal conversion efficiency of 60.78 % at 808 nm were applied to the construction of thermal analysis immunochromatographic test strips for the highly sensitive quantitative detection of aflatoxin B 1 (AFB 1 ) in grain. The CSA was coupled with the AFB 1 antibody to form a photothermal sensor probe by physical adsorption. The constructed immunosensor exhibited high sensitivity and a wide linear range from 0.01 to 10 μg/L in PBS. The detection limits of 0.00842 μg/L based on the thermal analysis was significantly improved by 11.88-fold compared with colorimetric results. No cross-reaction with the other mycotoxins was found except for aflatoxin B 2 , aflatoxin M 1 , aflatoxin G 1 and aflatoxin G 2 . Applied to analysize grain sample, the method achieved the detection of AFB 1 ranging from 0.16 to 160 μg/kg., 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 © 2022 Elsevier Ltd. All rights reserved.)

The existence of aflatoxin M 1 (AFM 1 ) 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 AFM 1 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 AFM 1 (490.30 ng/L) in comparison to other milks. AFM 1 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 AFM 1 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 AFM 1 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 AFM 1 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 AFM 1 in milk.

Aflatoxins (AFs) frequently contaminate food and animal feeds, especially in (sub) tropical countries. If animals consume contaminated feeds, AFs (mainly aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2) and their major metabolites aflatoxin M1 (AFM1) and M2 (AFM2)) can be transferred to edible tissues and products, such as eggs, liver and muscle tissue and milk, which ultimately can reach the human food chain. Currently, the European Union has established a maximum level for AFM1 in milk (0.05 µg kg -1 ). Dietary adsorbents, such as bentonite clay, have been used to reduce AFs exposure in animal husbandry and carry over to edible tissues and products. To investigate the efficacy of adding bentonite clay to animal diets in reducing the concentration of AFB1, AFB2, AFG1, AFG2, and the metabolites AFM1 and AFM2 in animal-derived foods (chicken muscle and liver, eggs, and cattle milk), chicken and cattle plasma and cattle ruminal fluid, a sensitive and selective ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method has been developed. High-throughput sample preparation procedures were optimized, allowing the analysis of 96 samples per analytical batch and consisted of a liquid extraction using 1% formic acid in acetonitrile, followed by a further clean-up using QuEChERS (muscle tissue), QuEChERS in combination with Oasis ® Ostro (liver tissue), Oasis ® Ostro (egg, plasma), and Oasis ® PRiME HLB (milk, ruminal fluid). The different procedures were validated in accordance with European guidelines. As a proof-of-concept, the final methods were used to successfully determine AFs concentrations in chicken and cattle samples collected during feeding trials for efficacy and safety evaluation of mycotoxin detoxifiers to protect against AFs as well as their carry-over to animal products.

Aflatoxin M 1 (AFM 1 ) is an important risk factor threatening the safety of milk and dairy products due to its carcinogenic and teratogenic effects on humans. To prevent AFM 1 from causing damage to human health, developing reliable methods to monitor its levels in milk and dairy products is of great importance. Biosensors built with recognition and detection systems have attracted extensive attention for their simplicity, portability, sensitivity, and selectivity. The commonly developed biosensors for detecting AFM 1 are antibody-based sensors (immunosensors) and aptamer-based sensors (aptasensors). This review focused on the advances of immunosensors, aptasensors, and other emerging receptors-based sensors for the detection of AFM 1 in milk and dairy products in the past five years. These biosensors were reviewed with representative examples according to their signal transduction systems, mainly including colorimetry, fluorescence, electrochemistry, and others. The unique advantages and drawbacks of immunosensor and aptasensor, and the future opportunities and challenges were also discussed., 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 © 2022 Elsevier Ltd. All rights reserved.)

Aflatoxins are potent carcinogenic and immunomodulatory mycotoxins, and exposure may lead to deleterious effects on human health. This study aimed to detect aflatoxin M 1 (AFM 1 ) as biomarker of exposure and determine associated risk factors in children attending a specialized-childcare hospital in Lahore. Urine samples collected from 238 children (1-11 years) during winter (January-mid-March 2020) and hot-humid summer (August-September 2020) were tested for AFM 1 presence using ELISA. Data on potential risk factors were also collected. Of 238 samples, 156 (65.5%) were positive for urinary AFM 1 . Season was significantly associated (OR = 2.64; 95% CI = 1.49-4.79; p  = 0.001) with AFM 1 positivity; prevalence was higher in hot-humid months (74.6%) than winter (57.3%). The place of living was also significantly associated (OR = 2.21; 95% CI = 1.25-3.97; p  = 0.007), and urinary AFM 1 positivity was higher in urban children (71.1%) compared to rural (58.3%). Median value for creatinine-adjusted AFM 1 was 1.9 ng/mg creatinine (Q1-Q3 = 0.82-6.0 ng/mg creatinine), while non-creatinine-adjusted AFM 1 was 0.57 ng/mL (Q1-Q3 = 0.23-1.4 ng/mL). Significantly higher urinary AFM 1 levels were detected in children; age ≤2 years ( p  = 0.037), who consumed more milk ( p  = 0.048), and who presented to the nutrition clinic ( p  = 0.003). These findings highlight the need for an effective control program to reduce the AFM 1 burden in children.

Aflatoxins are natural toxicants produced mainly by species of the Aspergillus genus, which contaminate virtually all feeds and foods. Apart from their deleterious health effects on humans and animals, they can be secreted unmodified or carried over into the milk of lactating females, thereby posing health risks to suckling babies. Aflatoxin M1 (AFM1) is the major and most toxic aflatoxin type after aflatoxin B1 (AFB1). It contaminates human breast milk upon direct ingestion from dairy products or by carry-over from the parent molecule (AFB1), which is hydroxylated in the liver and possibly in the mammary glands by cytochrome oxidase enzymes and then excreted into breast milk as AFM1 during lactation via the mammary alveolar epithelial cells. This puts suckling infants and children fed on this milk at a high risk, especially that their detoxifying activities are still weak at this age essentially due to immature liver as the main organ responsible for the detoxification of xenobiotics. The occurrence of AFM1 at toxic levels in human breast milk and associated health conditions in nursing children is well documented, with developing countries being the most affected. Different studies have demonstrated that contamination of human breast milk with AFM1 represents a real public health issue, which should be promptly and properly addressed to reduce its incidence. To this end, different actions have been suggested, including a wider and proper implementation of regulatory measures, not only for breast milk but also for foods and feeds as the upstream sources for breast milk contamination with AFM1. The promotion of awareness of lactating mothers through the organization of training sessions and mass media disclosures before and after parturition is of a paramount importance for the success of any action. This is especially relevant that there are no possible control measures to ensure compliance of lactating mothers to specific regulatory measures, which can yet be appropriate for the expansion of breast milk banks in industrialized countries and emergence of breast milk sellers. This review attempted to revisit the public health issues raised by mother milk contamination with AFM1, which remains undermined despite the numerous relevant publications highlighting the needs to tackle its incidence as a protective measure for the children physical and mental health.

In this present study, 195 cow milk, 100 goat milk, 50 buffalo milk, 50 camel milk, and 50 yak milk samples were collected in China in May and October 2016. The presence of aflatoxin M1 (AFM1) was determined using enzyme-linked immunosorbent assay method. For all cow milk samples, 128 samples (65.7%) contained AFM1 in concentrations ranging from 0.005 to 0.191 µg/L, and 6 samples (3.1%) from Sichuan province in October were contaminated with AFM1 above 0.05 µg/L (EU limit). For all goat milk samples, 76.0% of samples contained AFM1 in concentrations ranging from 0.005 to 0.135 µg/L, and 9 samples (9.0%) from Shanxi province in October were contaminated with AFM1 above 0.05 µg/L. For all buffalo milk samples, 24 samples (48.0%) contained AFM1 in concentrations ranging from 0.005 to 0.089 µg/L, and 2 samples collected in October were contaminated with AFM1 above 0.05 µg/L. Furthermore, 28.0% of samples contained AFM1 in concentrations ranging from 0.005 to 0.007 µg/L in camel milk samples, and 18.0% of samples contained AFM1 in concentrations ranging from 0.005 to 0.007 µg/L in yak milk samples. Our survey study has expanded the current knowledge of the occurrence of AFM1 in milk from five dairy species in China, in particular the minor dairy species.

A new green competitive ELISA for aflatoxin M1 quantification in raw milk was developed. This diagnostic tool is based on an anti AFM1 mAb produced by plant molecular farming in alternative to classical systems. Our assay, showing an IC 50 below 25 ng/L, fits with the requirements of EU legislation limits for AFM1 (50 ng/L). Optimal accuracy was achieved in correspondence of the decision levels (25 and 50 ng/L), and the assay enabled AFM1 quantification in the range 5-110 ng/L, with limit of detection 3 ng/L. Moreover, to evaluate a real applicability in diagnostics, raw milk-spiked samples were analysed, achieving satisfactory recovery rates of AFM1. In conclusion, an efficient and ready-to-use diagnostic assay for the quantification of aflatoxin M1 in milk, based on a plant-produced recombinant mAb, has been successfully developed.

Currently, there is a great demand for simple, sensitive, and accurate sensors for aflatoxin M1 (AFM1) in dairy products. In the present research, a novel fluorescent immunosensor based on nitrogen-doped carbon quantum dots (CQDs) has been developed for AFM1 analysis. The N-doped CQDs were synthesized through the hydrothermal approach using citric acid and polyethyleneimine as precursors. The CQDs showed bright blue emission under ultraviolet light irradiation and a maximum emission was observed at 450 nm upon excitation at 350 nm. The anti-AFM1 antibody (Ab) was immobilized on the as-obtained amine-functionalized CQDs and the obtained CQDs/Ab probe was then directly used for developing the immunoassays for AFM1. The fluorescence of the CQDs/Ab solution was effectively quenched in the presence of increasing AFM1 concentrations. Under the optimized conditions, the fluorescent nanosensor exhibited high sensitivity towards AFM1 in the range of 0.2-0.8 ng/mL with low limit of detection i.e., 0.07 ng/mL in standard buffer. Furthermore, the CQDs/Ab immunosensor was developed as a lateral flow design for detecting the aflatoxin residues in milk. This strategy can be used for the development of low-cost, rapid, and highly sensitive sensor strips for the detection of AFM1 in dairy products., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The simultaneous detection of three kinds of small-molecule contaminants (antibiotics, mycotoxins, and hormones) in milk was realized by using an 8-17 DNAzyme-based fluorescent enzyme-linked immunosorbent assay (ELISA), in which 8-17 DNAzyme was utilized as the catalytic enzyme for amplifying the signal. Compared with the conventional ELISA in which horseradish peroxidase is used as the catalyzing factor, this 8-17 DNAzyme-based ELISA could achieve multicolor signal output with lower detection limits. The linearities for chloramphenicol, 17β-estradiol, and aflatoxin M 1 were in the range of 0.3 ng/mL-3 μg/mL, 3 ng/mL-3 μg/mL, and 3 pg/mL-3 ng/mL with quantitation limits of 0.3, 3, and 0.003 ng/mL, respectively. This proposed scheme demonstrated that the 8-17 DNAzyme might be an effective substitute for horseradish peroxidase in ELISA for the development of ultrasensitive and multicolor fluorescence immunoassay, which would stimulate the development of ELISA in a new orientation.

This study investigates the reduction of aflatoxin M1 (AFM1) in skim milk by using ultraviolet light at 254 nm and the effects of influencing factors on the efficacy including treatment time (min), depth of samples (mm), contamination level (μg L -1 ), stirring, temperature, and fat content in milk. The colour and pH of milk samples were measured to evaluate the influence of the treatment on these values. It was found that short-wave ultraviolet radiation (UVC) reduced up to 50% of AFM1 in milk after 20 min of treatment regardless of the initial AFM1 contamination level. Treatment time, depth of samples, and stirring were all found to significantly (P < 0.05) enhance the reduction of AFM1. The milk colour was affected but there was no influence on the pH of milk samples at any duration of UV exposure. It is concluded that UVC light treatment has the potential to reduce AFM1 in milk., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In this study, solid-phase extraction (SPE) combined with the dispersive liquid-liquid microextraction based on novel hydrophobic deep eutectic solvent (DLLME - DES) has been developed as an ultra-pre-concentration technique for the extraction of aflatoxin M 1 (AFM1) in milk-based baby food (MBBF) and infant formula milk (IFM) samples followed by HPLC combined with fluorescence detection (HPLC - FL). In addition, carcinogenic and non-carcinogenic risk assessment was performed by health-related risk factors including liver cancer risk (LCR), margin of exposure (MOE) and target hazard quotient (THQ) were calculated using the mean of AFM1 in different infant food samples. The results of the study showed that the mean of AFM1 was statistically significant different between various brands and types of IFM and MBBF. The results of the study showed that the percentage of positive samples higher than the allowable limit of AFM1 in 36 samples of domestic infant formula milk (DIFM), 24 samples of imported infant formula milk (IIFM), 36 samples of domestic milk-based baby food (DMBBF) and 18 samples of imported milk-based baby food (IMBBF) were 41.6, 12.5, 66.7 and 33.3%, respectively. In addition, estimated values for health risk-related factors including LCR, MOE and THQ indicated that for most infants less than one-year-old were higher than the acceptable levels. Based on the results, it can be concluded that the quality of IFM and MBBF consumed in Iran in terms of AFM1 is poor. Therefore, it is necessary to take appropriate measures to reduce the amount of AFM1 in DIFM and DMBBF, and in addition, the IIFM and IMBBF should be controlled qualitatively before supplying the market.