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Aflatoxin M1 (AFM1) is a hepatocarcinogenic and hydroxylated metabolite of aflatoxin B1, detected in milk and milk products.
The aim of our research was to determine the incidence and risk assessment of AFM1 through the consumption of yoghurt in Hamadan province of Iran.
Fifty yoghurt samples were gathered from various areas of Hamadan province, Iran and tested for AFM1 by ELISA technique. The estimated daily intake (EDI) and the liver cancer incidence of aflatoxin M1 were determined.
AFM1 was detected in 43 (86%) samples, (mean: 28.56 ng/kg; range: <5-98.65 ng/kg). The level of AFM1 in 9 (18%) samples was above the maximum tolerance limit (50 ng/kg). The AFM1 intake through yoghurt consumption in various population groups ranged from 0.016 to 0.032 ng/kg bw/day in mean consumers and 0.019 to 0.046 ng/kg bw/day in high consumers.
The AFM1 intake through yoghurt contributed a slight part from the overall incidence of liver cancer in the Iranian population. From the findings of the current study, it can be derived that although the high percentage of yoghurt samples in Iran proved to be contaminated with AFM1 contents, did not show a public health concern considering the European Commission (EC) and the Institute of Standards and Industrial Research of Iran (ISIRI) maximum limits.
Yoghurt is one of the most important and popular products among fermented dairy products, which has been used as food consumed by the world population for thousands of years. Yoghurt contains nearly all nutrients necessary to sustain life [
Aflatoxin M1 (AFM1) is one of the groups of mycotoxin produced by
Many countries have implemented regulations to control the amount of mycotoxins groups among food and agricultural products, especially for AFM1 in milk products. These regulations differ among several countries with respect to their economic considerations [
The immediate detection of contamination is one of the beneficial methods to control aflatoxin M1 [
The aim of the current survey was to evaluate the occurrence and risk assessment of exposure of AFM1 through the consumption of yoghurt in the Hamadan province of Iran.
The current research is a cross-sectional study. For this purpose, 50 yoghurt samples were randomly purchased from supermarkets located in popular markets in different parts of Hamadan province, Iran, from October 2017 to August 2018. Eventually, all samples were carried to the lab and kept in the refrigerator at 4 oC. All samples were analyzed for AFM1 before the expiration date of the samples. All procedures of study were approved by the Ethics Committee of Hamadan University of Medical Sciences with No. IR.UMSHA.REC.1396.617.
The quantitative measurement of AFM1 in samples was distinguished by competitive ELISA using AFM1 test kit (RIDASCREEN® AFM1 Art. No.: R1121, R-Biopharm, Darmstadt, Germany). The preparation of the yoghurt samples and AFM1 measurement were performed according to the method described by the kit manufacturer. The mean lower limit of detection (LOD) for AFM1 in yoghurt was 5 ng/kg.
The risk of AFM1 intake through yoghurt was carried out by the deterministic approach and calculated according to the following equation [
AFM1 intake (ng/kg bw/day) = the concentration of AFM1 in yoghurt (ng/kg) × 95th percentile (for high consumers) or mean (for mean consumers) of per capita yoghurt consumption (kg)/ body weight (kg) Equation 1.
95th percentile (for high consumers) or mean (for mean consumers) of per capital yoghurt consumption was obtained by food frequency questionnaires (FFQ), Before the study, FFQ was prepared and given to people with different age-sex groups to complete it during a month. The people were randomly selected. For all participants in FFQ study, written informed consent was obtained.
For risk assessment, AFM1 value in yoghurt samples in which the concentration of this mycotoxin was lower than LOD was considered as LOD [
The liver cancer incidence due to AFM1 consumption was estimated according to Equation 2 [
Liver cancer incidence (cancers/yr/108 persons) = AFM1 intake (ng/kg bw/day) × potency
Potency= 0.001 × (1-P) + 0.03 × P
In these equations, the liver cancer potency of AFM1 in individuals positive for negative hepatitis B surface antigen (HBsAgz +) and individuals negative for negative hepatitis B surface antigen (HBsAgz -) was considered as 0.03 and 0.001 cancers/year/ ng of AFM1/ kg body weight/ day in a population of 100,000 [
The concentrations of AFM1 in milk samples were analyzed by SPSS Statistics 16.0 for Windows. One-side t-test was applied to compare the mean concentration of AFM1 samples with the maximum acceptable amount of the ISIRI and European Union (50 ng/kg) regulation. Differences between values were considered significant at
The occurrence and levels of AFM1 in yoghurt samples consumed in Hamadan province are presented in Table
The AFM1 intakes through yoghurt in different age-sex groups of Iran population are shown in Table
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Yoghurt | 50 | 43 (86%) | 28.56 | 26.39 | 7 (14%) | 34 (68%) | 9 (18%) | <5-98.65 |
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Male | 56 | 27.56 | 0.025 | 0.026 | 0.039 | 0.034 | 0.036 | 0.054 | |
Female | 68 | 25.12 | 0.027 | 0.032 | 0.048 | 0.040 | 0.046 | 0.069 | |
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Male | 85 | 52.16 | 0.034 | 0.019 | 0.028 | 0.042 | 0.024 | 0.035 | |
Female | 68 | 48.15 | 0.037 | 0.023 | 0.034 | 0.045 | 0.028 | 0.041 | |
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Male | 95 | 78.65 | 0.042 | 0.016 | 0.023 | 0.051 | 0.019 | 0.028 | |
Female | 89 | 64.25 | 0.045 | 0.020 | 0.030 | 0.054 | 0.024 | 0.036 | |
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Male | 45 | 71.25 | 0.045 | 0.018 | 0.027 | 0.055 | 0.023 | 0.034 | |
Female | 53 | 59.36 | 0.046 | 0.023 | 0.034 | 0.058 | 0.029 | 0.043 |
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Iran | 61 | 30 (49.2) | TLC | 26 | 15 – 102 | 10 (16.4) | Fallah |
Turkey | 80 | 70 (87.5) | ELISA | 66.1 | 10 – 475 | 16 (20) | Atasever |
Iran | 60 | 59 (98.33) | ELISA | 51.66 | 6.2 – 87 | 38 (63.33) | Issazadeh |
Turkey | 26 | 26 (100) | ELISA | 238 | 125 – 269 | 26 (100) | Tosun |
Pakistan | 96 | 59 (61) | HPLC | 90.4 | 4 – 615.8 | 28 (47) | Iqbal |
Pakistan | 96 | 32 (33.33 ) | HPLC | 90.4 | LOD – 880 | 21 (21.87) | Iqbal |
Turkey | 50 | 50 (100) | ELISA | 55.28 | 40.62 – 72.04 | 5 (10) | Temamogullari & Kanici [ |
Iran | 60 | 48 (80) | ELISA | 130.5 | 19.7 – 319.4 | 3 (5) | Rahimi [39] |
Iran (Traditional) | 40 | 40 (100) | ELISA | 33.6 | 6 – 91 | 1 (2.5) | Mason |
Iran | 42 | 10 (23.8) | ELISA | 15.1 | 6.3 – 21.3 | 0 | Bahrami |
South Korea | 55 | 15 (27.27) | HPLC | 51* | 20 – 150 | NR | Kim-Soo |
Malaysia | 5 | 2 (40) | ELISA | 16.45* | 7.5 – 31 | 0 | Nadira |
Pakistan | 66 | 26 (39.39) | HPLC | 56 | LOD – 196.3 | 8 (12.12) | Iqbal |
Qatar | 21 | 16 (76.1) | ELISA | 31.32 | 4.16-38.21 | 0 | Hassan |
Kenya | 38 | NR | ELISA | 117 | 17 – 1100 | 25 (65.78 ) | Lindahl |
China | 27 | 15 (55.5) | ELISA | 17.2 | 4 – 47 | 0 | Guo |
Yoghurt is a favorite fermented dairy product, which is used as part of the popular diet in Iran because yoghurt is useful to affect human’s health and has nutritional value. Various types of fermented dairy products have been made and consumed in individual households in Iran, Turkey, Qatar, Lebanon, and other Middle Eastern countries for centuries.
Considering the present findings, we detected a high level of AFM1 contamination in yoghurt samples from Iran. In a prior survey, Cano-Sancho
Hassan and Kassaift [
The other obtained results reported by Tabari
In a previous study performed in Iran, Mason
According to several studies carried out in different neighboring countries of Iran, Turkey Altun
As seen in Table
The incidence of liver cancer in Iran was 3.53 cancers per year per 105 persons or 3530 cancers/yr/108 persons [
From our findings of the current study, it can be derived that although a high percentage of yoghurt samples in Iran proved to have AFM1 contents, but it does not show a public health concern considering the European Commission (EC) and the Institute of Standards and Industrial Research of Iran (ISIRI) maximum limits. However, regarding the important role of milk, especially dairy products in the human diet, there is a huge concern about the presence of AFM1 in milk and dairy products. Hence, it is important to use fast methods in the detection of AFM1 in milk and dairy products; and also, the Iranian public health authorities have to monitor ceaselessly to detect AFM1 contamination.
Ali Heshmati, and Amir Sasan Mozaffari Nejad conceived, designed, analyzed, and interpreted the data; Tayebeh Ghyasvand and Amir Sasan Mozaffari Nejad performed data collection. Ali Heshmati and Amir Sasan Mozaffari Nejad wrote the first draft and finalized it. All authors read and approved the final manuscript.
All procedures of the study were approved by the Ethics Committee of Hamadan University of Medical Sciences, Iran with No. IR.UMSHA.REC.1396.617.
Not applicable.
Not applicable.
The data that support the findings of this study are with the corresponding author, [ASMN], and can be made available on reasonable request.
The authors appreciate the Vice-Chancellor of Research and Technology of Hamadan University of Medical Sciences for financial support (Project No. 9609286051).
The authors declare no conflicts of interest, financial or otherwise.
Declared none.