AFM1 Contamination Research Articles

Aflatoxin M1 in raw milk in summer season in Lahore, Pakistan

A four-month long study throughout the summer of 2022 i.e., April, May, June, and July; was conducted to determine Aflatoxin M1 concentration in raw milk retailed in the shops of Lahore, Pakistan. A total of 40 samples (10 per month) were analyzed using the ELISA kit method while considering European Union (50ng/kg), US Food and Drug Regulation Authority (500ng/kg), and Pakistan Pure Food Regulations (500ng/kg) as AFM1 permissible limits. About 82.50% of 40 samples were contaminated with AFM1 while only 17.50% samples were found to be uncontaminated. Of these contaminated samples over 87.88% samples exceeded the safe limit as set by the European Union i.e., 50ng/kg with only 12.12% of the samples being safe for use. On the other hand, only 6.06% of the samples were unfit as per the USFDA & Punjab Pure Food Regulations (PPFR) limit i.e., 500ng/kg. A mean value of 133.57 ± 0.14ng/kg of AFM1 contamination was calculated ranging from 5.43 ± 0.05ng/kg to 964.75 ± 0.04ng/kg in April and July, respectively. The lowest average contamination was recorded in May i.e., 68.35 ± 0.258ng/kg which went progressively higher in the later months due to unusual record-breaking rainfall in the region while highest average contamination was found in the month of July i.e., 228.789 ± 0.075ng/kg. The current study is significantly important as it identify the contamination of Aflatoxin M1 in raw milk in the summer season. Bangladesh J. Sci. Ind. Res. 59(2), 115-122, 2024

Occurrence of Aflatoxin M1 and Estimate of Dietary Exposure in Cheeses from Organic and Conventional Production Systems

This study aimed to compare AFM1 occurrence in different cheese types produced by organic and conventional systems; and to evaluate the risk of food exposure to AFM1. A total of 176 commercial cheeses of 17 types were analyzed, 84 of organic and 92 of conventional production. Determination of AFM1 was performed by high- performance liquid chromatography (HPLC), being detected in 30.5% of samples, with 4.8% of organic cheese samples presenting quantifiable AFM1 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 AFM1 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 AFM1 (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 AFM1 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 AFM1 is an obstacle to dairy production. Control and prevention of AFM1 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.

Detection of Aflatoxin M1 and Several Heavy Metals in Medical Infant Milk Formula Sold in Iraqi Markets

Milk is essential for infants, and infant feeding should be emphasized in any program to ensure good child health. To find harmful and trace elements in different milk powder brands bought in Iraq and fed to babies with metabolic syndrome, atomic absorption spectrometry (AAS) was used. Fluorescence high-performance liquid chromatography (HPLC) was also used to measure aflatoxin M1. The result reveals the concentrations of elements close to the product label's description, such as magnesium, calcium, zinc, selenium, iron, and manganese. Only the potassium element was higher than what was listed on the product label. Additionally, no quantities of hazardous elements like lead (Pb), cadmium (Cd), or arsenic (As) were found. As for aflatoxin M1, it was found that the PKU milk formula and lactose-free milk formula were found to be free of AFM1 contamination, indicating that they are safe to consume. AFM1 was found in concentrations above the permissible limit in the MSUD milk formula (0.08 µg/kg). The reason for this issue could be inadequate storage. The other two types, TYR and OAc milk formula, were lower than the allowable limit of 0.05 µg/kg. As a result, there needs to be more oversight of the milk that newborns with metabolic syndrome consumele S. aureus or Streptococcus pyogenes, and Streptococcus pneumoniae in community settings.

Bio-transference of Aflatoxin B1 from feed to the breast of hens as Aflatoxin M1

The presence of hydroxylated metabolites of aflatoxin B1 (AFB1), such as M1 (AFM1) Aflatoxin has been reported in breast laying hen, Diaz Zaragoza et al, 2014. The hens when their laying cycle end are sent to the trail of bird for his sacrifice. In Mexico chicken consumption in the form of stock, it is much in demand mainly in the center and southeast of the country. Some companies process meat to prepare various types of sausages, such that in one way or another becomes a food for human consumption. Currently the System Product Poultry in Mexico, is working to raise awareness about food safety in the poultry industry generated as egg and meat. The AFM1 is a compound considered as a carcinogen and is limited in milk at levels of 0.5 to 0.05 µg/L. This depend the regulations of each region of the world. The given the possibility that AFM1 is a chemical risk consumption of hen breast. Evaluating the Bio-transfer this AFB1 was performed in a contaminated diet with 500 µg/kg (ppb) and quantify AFM1 contamination in the breast, after 8 days of diet consumption. At the same time included an anti-aflatoxin agent to verify their efficiency in this experiment. AFM1 concentration in the breast of the negative control group was below the detection limit of 0.001 ppb. The concentration of the positive control group was 0.004 ppb and challenge group concentration was 0.002 ppb. The analytical tests were carried out by the technique of Ultra High Performance Liquid Chromatography (UHPLC). The results show that bio-AFB1 transfer to the breast as AFM1 is minimal. Therefore, this food is considered safe. Evaluated adsorbent efficiency was 50%. Complementarily AFB1 concentration in the liver, where the toxin is quantified levels of 0.467 ppb in the positive control and 0.196 ppb in the challenge group with adsorbent was evaluated. The effectiveness of the commercial adsorbent was the 60%.

Determination Of Aflatoxin M1 In Raw Cow's Milk Using Hplc- Fld, In Injibara Town, Awi Zone, Amhara, Ethiopia

Background: Consumption of contaminated milk is a major health problem for consumers, and aflatoxins are potent liver carcinogens, mutagens, and immunosuppressive compounds. Therefore, raw cow milk must be constantly monitored for the presence of AFM1 contamination. It is extremely important to keep AFM1 levels low in raw cow’s milk. The level of AFM1 in milk varies depending on the level of AFB1 ingested by dairy cattle through the feed. Objective: This investigation is mainly focused on the detection of levels of AFM1 in raw cow’s milk. There is little information on the level and distribution of aflatoxin in raw milk samples from cows in the vicinity of Injibara Town. Therefore, it is important to analyze the level and distribution of aflatoxin in raw milk samples from cows in Injibara Town. Methods: This study is mainly a quantitative investigation of the amount of AFM1 in raw cow’s milk using the HPLC-FLD spectrometry methods. All the milk samples were collected using a random sampling technique method from different areas during the night and morning a day and the collected raw cow’s milk samples were stored in a clean polyethylene bag. Results: In the present study, a total of 324 raw cow’s milk samples were collected from farmers in Injibara town, and samples were analyzed in triplicate for AFM1 determination by HPLC-FLD. Aflatoxin M1 was detected in 3 (0.92 percent of whole samples) out of 324 samples with quantities ranging from 0.046 to 0.22 µg/L and a mean value of 0.121 µg/L. From 324 raw cow’s milk samples, two (0.62 %) were seen over and one sample (0.31 %) was below the permissible limits of EU and ES (0.05 µg/L. The most number of samples were not detected in AFM1 analysis. Conclusions: From this result conclude that two samples (0.096 and 0.22 μg/L) were over the EU (0.05μg/L) and ES (0.05μg/L) permitted levels and under analysis, one sample (0.046 μg/L) was below the standard limit in the USA (0.5 μg/L). The average contamination was 0.121µg/L and this data revealed that the raw cow’s milk is to a small extent contaminated with aflatoxins.

Analysis of the Aflatoxin M1 contamination in traditional and commercial cheeses consumed in Yemen

The current study aimed to determine the levels of Aflatoxin M1 in traditional Taizi cheese and commercial cheeses utilised in Yemen. A total of 273 cheese samples were collected from supermarkets and analysed for AFM1 contamination by enzyme‐linked immunosorbent assay (ELISA) and high‐performance liquid chromatography (HPLC). Among these samples, 79.12% were positive for AFM1 contamination with levels ranging from 0.020 to 5.955 μg/kg. AFM1 was detected at 84.67 and 73.53% in commercial and traditional cheeses, respectively. In the current study, 32.6% of the total cheese samples were contaminated with AFM1 at a level above the acceptable limit (0.250 μg/kg) of the European Commission. Results of this study revealed that the presence of AFM1 contamination in different types of cheese samples should be considered a serious public health problem in Yemen.

Presence of Mycotoxins in Feed and Dairy Products of Cattle in Paraná, Brazil

Milk contamination by mycotoxins is considered a public health problem. Therefore, the objective of this study was to identify these contaminants in concentrates and in the milk from 31 high-producing herds. Only two of the 55 concentrate samples analyzed showed the presence of aflatoxin G1 (AFG1, 3.2 and 3.6 μg·kg-1). AFM1 was detected in 93.5% of the 62 milk samples analyzed with a range from 0.045 to 0.442 μg·L-1. All of the AFM1 concentrations were below the maximum limit tolerated (0.5 μg·kg-1) by the Brazilian Agência Nacional de Vigilância Sanitária (ANVISA). There was no difference in AFM1 contamination levels for both sampling periods (summer and winter). In conclusion, the AFM1 contamination in animal feed and, consequently, in milk is within the limits tolerated by ANVISA for Brazil (≤ 0.5 μg·kg-1). Furthermore, no season effect on AFM1 levels was found.

Assessment of aflatoxin in milk and feed samples and impact of seasonal variations in the Punjab, Pakistan.

The present study was designed to assess the incidence of aflatoxin contamination in animal feed and raw milk samples (total 240 each) collected from dairy farms during the complete year of 2015. These samples were collected through a cluster random sampling technique by dividing the province of Punjab, Pakistan into five clusters (north, south, east, west and central). Factors (environmental & physical) affecting aflatoxin contamination in milk and animal feed at farms were also studied. The AFM1 levels in raw milk & AFB1 levels in feed samples were analyzed by using the ELISA technique. Results demonstrated that overall about 53% raw milk samples from dairy farms were contaminated beyond the US MRL (0.50 µg/L) for AFM1 with than average level of 0.59 µg/L, while the 95% farm feed samples were exceeding the FDA MRL (20 µg/kg) of AFB1 with average level of 43 µg/kg. During winter season, the concentration of AFM1 was higher in all clusters with avg 0.68 µg/L, while the AFB1 contamination was highest in the spring season with avg 54 µg/kg. Market feed prices had negative correlation with AFB1 contamilevels, which were further supported by the positive correlation between quantity of feed at farms with AFM1 and AFB1 contamination. Results exhibited significantly positive impact of environmental factors on milk and feed aflatoxin contamination levels, whereas temperature showed an inverse relationship with AFM1 and AFB1 levels. The study recommends need of synergistic extension work to support dairy farms and highlight the contamination levels for regulatory bodies to introduce strategic policies for control measures.

Aflatoxin Contamination of Milk Produced in Peri-urban Farms of Pakistan: Prevalence and Contributory Factors.

Aflatoxin M1 contamination of milk in Pakistan, like many developing countries, is poorly understood. The present study was therefore conducted to determine AFM1 contamination of milk and its contributory factors in Pakistan. We sampled milk and feedstuffs from 450 peri-urban dairy farms in seven major cities following a cross-sectional study design. Analysis of milk using ELISA revealed high contamination with an overall average of 3164.5 ng of AFM1/L, and significant differences (p < 0.001) between cities. The milk sampled from Gilgit, in northern hilly areas, had an average AFM1 level of 92.5 ng/L. Milk from other cities had 3529.7 ng/L average contamination, with only 5.7% samples qualifying the maximum tolerable limit of 500 ng of AFM1/L. Heavy mean aflatoxin contamination was found in bakery waste (724.6 μg/kg), and cottonseed cake (600.8 μg/kg). Rest of the other feedstuffs had moderate to low mean aflatoxin contamination, ranging from 66.0 μg/kg in maize stover to 3.4 μg/kg in wheat bran. The mean aflatoxin level in commercial dairy concentrates was 32.7 µg/kg. About 80% of the total aflatoxin intake of dairy animals was contributed by cottonseed cake alone due to its high aflatoxin contamination and proportion in dairy rations. On-farm storage time of oilseed cakes varied (p < 0.01) in different cities but was not associated with aflatoxin contamination. The exceptionally high AFM1 contamination suggests that milk from peri-urban dairy farms is a serious public health threat in Pakistan. This situation can be mitigated by reducing aflatoxin contamination in cottonseed cake and promoting the use of commercial concentrates and other feedstuffs with low contamination.

Reduction of Milk Contamination with Aflatoxin-M1 through Vaccination of Dairy Cattle with Aflatoxin-B1 Vaccine

Aflatoxin M1 is one of mycotoxin derivatives, which is secreted in milk of dairy cattle fed on feed contaminated with Aflatoxin-B1 (AFB1). The current study was designed to prepare a vaccine against AFB1and to evaluate its efficacy in reducing or preventing secretion of AFM1 in milk. Aflatoxin-B1 was prepared, purified and transformed into oxime, then it was fixed on bovine serum albumins. The AFB1-BSA conjugate was adjuvanted with Gold Nano particles then Montanide ISA 206. The prepared vaccine was used for immunization of rabbits by S/c routes as 100 µg/dose and dairy cattle by I/M routes as 500 µg/dose. The vaccinated animals were boosted at 3 weeks post primary immunization. Serum samples were collected and examined for the anti-AFB1 using AGPT. A mean titer of 15.2 AGPU/ml was detected at 2 weeks post primary vaccination then significantly increased till reached to 76.8 AGPU/ml at 6 weeks post Booster vaccination. All vaccinated rabbits were challenged with dose of 0.3 mg AFB1 toxin/Kg. The vaccinated rabbit showed 100% protection and no AFB1 toxin residue was detected in their livers. Milk samples were collected from non-vaccinated and AFB1-immunized dairy cattle then examined with ELISA for quantitation of AFM1 residues before and after vaccination. The results showed that the prepared AFB1 vaccine was safe, potent and able to reduce AFM1 release in milk of vaccinated heifers by 70%. So the vaccination of lactating animals with the AFB1vaccine might represent a valid tool for the prevention of AFM1 contamination of milk and dairy products.

Aflatoxin M1 in Nicaraguan and locally made hard white cheeses marketed in El Salvador

Aflatoxin M1 is a carcinogenic and genotoxic metabolite of Aflatoxins present in food contaminated by fungi for lactating cattle, it is excreted through milk and when used to make cheese, the toxin will also be transferred to the dairy. The contamination of unripened hard white cheese with AFM1 seems to vary according to the season of the year, possibly due to the change of foodstuff, from fresh pasture in the rainy season to dried foods in the dry season and vice versa. This research determined both the prevalence and contents of AFM1 in cheeses of local and Nicaraguan origin marketed in El Salvador, as well as the changes occurred according to the season and the association between levels of AFM1 with meteorological parameters. The significantly higher prevalence of AFM1 contamination in both local cheeses and Nicaraguans, was found in the dry season and the lowest in the rainy season (41 % vs. 20 %; 31 % vs. 0%, respectively), the same trend was observed in AFM1 contents (0.076 vs. 0.036 μg/kg; 0.050 vs. 0.021 μg/kg, respectively). A significant association was demonstrated between levels of AFM1 with the averages of accumulated rainfall and relative humidity according to the sampled season. The prevalence of AFM1 in cheeses indicate that El Salvador and Nicaragua are endemic to dairy contamination by that mycotoxin. Seasonal variation may be due to a lack of rainfall, that promotes the growth of aflatoxigenic fungi in the crops of raw materials, which will be used for feedstuff intended for dairy cattle, thus, the consumption of contaminated food will cause the temporary increase of AFM1 in milk and their derivatives.

Occurrence of Aflatoxin M1 in Raw Milk Marketed in Italy: Exposure Assessment and Risk Characterization.

The current study is based on the AFM1 contamination of milk determined from April 2013 to December 2018 in the framework of a self-control plan of six milk processing plants in Italy. These data – together with the consumption data of milk consumers – were evaluated and used for the calculation of the Estimated Daily Intake (EDI), the Hazard Index (HI), and the fraction of hepatocarcinoma cases (HCC) due to AFM1 exposure in different population groups. Altogether a total of 31,702 milk samples were analyzed, representing 556,413 tons of milk, which is an outstanding amount compared to published studies. The results indicate the monthly fluctuation of AFM1 levels through a period of nearly 6 years. The EDI of AFM1 in different population groups was in the range of 0.025–0.328 ng kg−1 body weight (bw) per day, based on the average consumption levels and weighted mean contamination of the milk in the study period. Considering average consumptions, in the groups of infants and toddlers, the HI calculation resulted in 1.64 and 1.4, respectively, while for older age groups, it was <1. The estimated fractions of HCC incidences attributable to the AFM1 intakes were 0.005 and 0.004 cases per 100,000 individuals in the 0–0.9 and 1–2.9-year age groups, respectively, and below 0.004 cases in the other age categories. The monthly average AFM1 contamination of tested milk consignments ranged between 7.19 and 22.53 ng kg−1. Although the results of this extensive investigation showed a low risk of HCC, the variability of climatic conditions throughout years that influence AFB1 contamination of feed and consequently AFM1 contamination of milk justifies their continuous monitoring and update of the risk assessment.

Assorted Methods for Decontamination of Aflatoxin M1 in Milk Using Microbial Adsorbents.

Aflatoxins (AF) are carcinogenic metabolites produced by different species of Aspergillus which readily colonize crops. AFM1 is secreted in the milk of lactating mammals through the ingestion of feedstuffs contaminated by aflatoxin B1 (AFB1). Therefore, its presence in milk, even in small amounts, presents a real concern for dairy industries and consumers of dairy products. Different strategies can lead to the reduction of AFM1 contamination levels in milk. They include adopting good agricultural practices, decreasing the AFB1 contamination of animal feeds, or using diverse types of adsorbent materials. One of the most effective types of adsorbents used for AFM1 decontamination are those of microbial origin. This review discusses current issues about AFM1 decontamination methods. These methods are based on the use of different bio-adsorbent agents such as bacteria and yeasts to complex AFM1 in milk. Moreover, this review answers some of the raised concerns about the binding stability of the formed AFM1-microbial complex. Thus, the efficiency of the decontamination methods was addressed, and plausible experimental variants were discussed.

The Role of Probiotic Bacteria in Protecting against Aflatoxin M1 Contamination in Milk and Certain Dairy Products

Purpose:The present study investigated AFM1 contamination in raw milk, kareish cheese and yoghurt and studied the abilities of heating and probiotic bacteria to reduce AFM1 contamination in both raw milk and yoghurt. Methods: AFM1 was detected in raw milk in 10 of 12 samples (83.3%) from each season. Two methods were used to assess the toxigenic potential. HPTLC was performed for aflatoxin detection. Aflatoxin M1 contamination milk and selected dairy products was investigated against heating and Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 and its combination. Result: Cheese had the highest mean values concentration of AFM1 (527.4 and 651.3 ng/kg), and yoghurt had the lowest concentration (39.13 and 64.68 ng/kg) while, raw milk samples were (207.0 and 311.8 ng/kg) during summer and winter, respectively. The concentration of toxin in 83.33% of the examined cheese samples exceeded the EU limit (50 ng/kg), and 70.83% of these samples exceeded the Codex limit (500 ng/kg) during both seasons. Boiling degraded 26.71% of the AFM1. Whereas, pasteurization only degraded 15.45%. In yoghurt, two strains of probiotic bacteria (Lactobacillus acidophilus and Bifidobacterium lactis) each gradually reduced the AFM1 concentration as a function of time in milk contaminated with 25 ng/l, with complete elimination by the end of the refrigerated storage period (3 days), while, the combination of both probiotic bacteria (L. acidophilus and B. lactis) was better able to reduce AFM1 in milk contaminated with 50.0 ng/kg or 75 ng/kg AFM1.Conclusion: The most extensive reductions of the AFM1 concentration were to 41.80 ng/kg (45.3%), 22.6 ng/kg (69.90%) and 7.12 ng/kg (92.8%), which were achieved using the same concentration of each strain individually and in combination, respectively, after two days in milk contaminated with 75 ng/kg. No AFM1 was detected after three days using the combined strains.

Diyarbakır’da Tüketime Sunulan Süt ve Ürünlerinde Aflatoksin M1 düzeylerinin ELISA ile Belirlenmesi Üzerine Bir Araştırma

In this study, the contamination levels of Aflatoxin M1 of 248 sterilized/UHT milk, Beyaz Cheese, Cecil Cheese, Cokelek, Cream Cheese, Lor Cheese, Van Otlu (herbed) Cheese, Van Otlu Lavash Cheese and butter samples from randomly selected market and supermarkets from Diyarbakir province center were determined by ELISA method. The average AFM1 contamination value of the cheese and butter samples investigated was 133.51 ng/kg; while the minimum and maximum values were 50 and 595.31 ng/kg, respectively. On the other hand, the average AFM1 value of 13 sterilized milk samples was 52.59 ng/kg; minimum and maximum values were determined as 25.97 and 80.0 ng/kg, respectively. In the study, the AFM1 results of 34 samples (13.7%) of sterilized milk, Cecil Cheese, Cokelek, Cream Cheese, Lor Cheese, Van Otlu Cheese, Van Otlu Lavash Cheese and butter samples were exceeded the limits set by the European Commission for AFM1 (50 and 250 ng/kg). The results of the analysis show that the milk and dairy products those were offered for consumption in Diyarbakir had a limited potential for consumers in terms of AFM1.

A survey of AFM1 contamination in row cow milk produced in region of Kumanovo

Aflatoxins (AFs) are type of mycotoxins produced by certain Aspergillus species. In the livers of cows, the aflatoxin В1 (AFB1) ingested with contaminated food is metabolized in carcinogenic aflatoxin М1 (AFM1) which is then excreted in the milk and poses a serious risk to the health of the consumers. In January of 2019, twenty five (25) samples of milk were collected from local farms in the surrounding area of Kumanovo, Republic of North Macedonia. The samples were analysed in the Centre for Public Health, Kumanovo. A method MKC EN ISO 14501:2007 for analysis of AFM1 by HPLC with fluorescence detection was applied. This method is selective, precise and applicative. The final analytical result has shown AFM1 below the LOD in any of the analyzed samples. Due to the well-known fact that AFM1 is toxic, i.e. carcinogenic and the fact that milk is widely used as staple food, continuous development as well as introduction of new, more sensitive methods for detection of AFM1 are required. Keywords: Aflatoxin M1, HPLC-FLD, milk

Detection of Aflatoxin Contamination of Dairy Production Chain in the Northwest Region of Rio Grande do Sul, Brazil

The present study evaluates levels of aflatoxin contamination involving small dairy farms from the in the Northwest region of Rio Grande do Sul, Brazil. The objectives of this trial is: quantify lactating cow diet and raw for bulk milk Aflatoxin B1 (AFB1) contamination; evaluate the use of a mycotoxin adsorbent on cows submitted to semi-confinement raising; and verify the presence of AFM1 contamination on UHT and powdered milk commercialized locally. For this, 17 farms were selected. During the experimental period, every lactating cow on each herd received 50 g/day of a commercial mycotoxin adsorbent for dairy cows (100% bentonite). From each farm, bulk milk, corn silage and concentrate samples were collected for aflatoxin quantification. Aflatoxin M1 was detected in all milk samples examined from each farm, but the levels were within the allowed by the legislation. Aflatoxin B1 was also detected in all samples of corn silage and concentrate. The mycotoxin adsorbent used reduced (p &lt; 0.05) AFM1 contamination in milk. AFM1 milk contamination levels observed on commercialized milk also respected Brazilian requirements. It is possible to conclude that there is a high incidence of Aflatoxin B1 contamination on corn silage and dairy cows concentrates in family farms from South Brazil region. Based on the condition in which the present study was conducted, the use of a mycotoxin adsorbent reduced the levels of AFM in milk in production system. However, the locally produced milk and commercial milk were below the AFM levels accepted by the National Requirements for AFM1 contamination.

The Incidence of Aflatoxin M1 in Milk Produced in a Regional Surveillance System Between 2013 and 2017

Aflatoxins are toxic metabolites, generally produced by Aspergillus flavus, A. parasiticus and A. Nomius. They can have immunosuppressive, mutagenic, teratogenic and carcinogenic effects, especially on the liver. The purpose of this study was to evaluate the frequency of AFM1 contamination in milk in an established surveillance area of Romania, during 2013-2017 period. The material was represented by 150 raw milk samples obtained from milk collecting units and breeding units in Cluj county. The samples were analyzed by ELISA method. Measurable levels of AFM1 were present in 28% of the milk samples examined. The level of AFM1 in the positive samples ranged between 0,010 and 0,089 μg/kg. Of the total samples, only two were found non-compliant with the EU regulatory limit (0.05 μg/kg). All the positive samples were detected in 2013. In the 2014-2017 period, the level of aflatoxin M1 in milk was undetectable. The contamination risk of AFM1 in milk may increase with feeds being stored in improper conditions of humidity and temperature. This risk can be significantly reduced by using aflatoxin uncontaminated feeds.

Occurrence of aflatoxin M1 in milk samples from Italy analysed by online-SPE UHPLC-MS/MS

The occurrence of aflatoxin M1 in 69 milk samples collected in a south region of Italy in 2016 was evaluated. The samples were analysed using an automated method based on online SPE coupled with UHPLC tandem mass spectrometry. After a salt induced liquid–liquid extraction with acetonitrile to remove protein from milk, the extract was diluted with water and analysed using an automated online SPE MS/MS method. Among the analysed samples no one had AFM1 higher than the legally allowable limits whereas 71.4% of the other analysed samples were above the LOD of the method. The highest contamination level of AFM1 was found in pasteurised milk (44.39 ng kg−1). The results show the worrying and widespread of AFM1 contamination, highlighting the necessity of monitoring studies in order to evaluate the reduction of the maximum legal limit.

Aflatoxin M1 Contamination Levels in Cheeses Sold in Isfahan Province, Iran.

ObjectivesAflatoxin M1 (AFM1)-contaminated dairy products pose serious human health risks, causing liver and renal failure if consumed. They are also related to decreased milk and egg production in infected animals. This study investigated the AFM1 contamination levels in cheeses sold in Isfahan province, Iran, by enzyme-linked immunosorbent assay (ELISA).MethodsA total of 100 white cheese samples were randomly collected from supermarkets in Isfahan province and after extraction using dichloromethane were prepared for the ELISA.ResultsOf the 100 samples, 52 (52%) were contaminated by AFM1, at levels ranging from 50.2 to 424.4 ng/kg. The remaining 48% of the samples had undetectable AFM1 levels (< 50 ng/kg). Based on the standard limit set by the European Commission and Iran, 8% (8/100) of the AFM1-positive samples (with concentrations between 250.2 and 424.4 ng/kg) had levels higher than the permissible value of 250 ng/kg.ConclusionAlthough the percentage of cheese samples in Isfahan province with AFM1 levels exceeding the national permissible limit was low, the examination of cheeses and the milk used for their production is nevertheless important for ensuring public health. Furthermore, optimum storage conditions of animal feed should be ensured, and livestock nutrition must be monitored for the presence of AFM1 and other aflatoxins.