Determination of aflatoxin M1 in breast milk and related factors

SUMMARY OBJECTIVE: Breastfeeding in women with aflatoxin M1 exposure may be a risk factor for the newborn. Thus, it is crucial to determine aflatoxin M1 levels in breast milk and raise mothers' awareness about nutrition in lactation and other periods. This study was carried out to determine aflatoxin M1 contamination in milk samples taken from mothers who gave birth. METHODS: The study was carried out in the postpartum department of Training and Research Hospital between December 31, 2018, and June 31, 2019, and 90 breastfeeding mothers were included in the study. RESULTS: A total of 75 (83.3%) of the examined samples were found positive. The mean aflatoxin M1 ratio in positive samples was 12.16 pg/mL (5.00-23.18 pg/mL). Mothers' consumption of processed food was associated with aflatoxin M1 levels (p=0.043). It was determined that the aflatoxin M1 levels of mothers who consumed processed food products 1 or 2 times a month were 3.22 times lower than those who consumed 1-2 times a week. CONCLUSIONS: This study emphasized the importance of monitoring aflatoxin M1 levels in breast milk for infant health. It is thought that nutrition education given to mothers during pregnancy will significantly impact aflatoxin M1 results. In addition, the dangers of mycotoxins in mother-infant nutrition should be emphasized regularly in health education.

Detección de aflatoxina M1 en muestras de leche cruda de vacas en tanques de enfriamiento en Boyacá (Colombia) / Detection of aflatoxin M1 in raw cow milk samples in cooling tanks in Boyacá (Colombia)

RESUMEN Una de las principales formas de contaminación de la leche con micotoxinas es el consumo de alimentos fermentados que se encuentran contaminados con mohos principalmente de Aspergillus spp., los cuales producen toxinas que pueden llegar a constituirse como un problema para la salud publica debido a su estabilidad térmica y química. El objetivo del presente trabajo fue detectar las concentraciones de aflatoxina M1 en muestras de leche de vacas en tanques de enfriamiento en cuatro municipios del departamento de Boyacá durante un año, determinando las variaciones de acuerdo con la temporada. Se realizó un estudio de corte longitudinal, descriptivo cuantitativo. Se seleccionaron aleatoriamente cuatro tanques de enfriamiento de cuatro municipios distintos del departamento; cada uno se muestreó dos veces al mes durante todo el período de estudio y se procesaron mediante metodología Charm Ez Lite . Se realizó un ANDEVA para determinar las diferencias estadísticas entre las concentraciones de la aflatoxina M1 por cada trimestre. Se determinaron diferencias estadísticas entre cada uno de los trimestres del estudio encontrando un porcentaje de positividad de 74,06% del total de muestras positivas en los trimestres de verano. 28,12% (108) de las muestras tomadas durante todo el estudio fueron positivas, con concentraciones de la toxina que oscilaron entre 0,5 y 2,0 μg/Kg de leche. Se determinó por primera vez en el departamento de Boyacá las concentraciones y variaciones estacionales de aflatoxina M1 en muestras de tanques de enfriamiento de leche, encontrando las mayores concentraciones y número de casos positivos de aflatoxina M1 en los meses de verano.

ABSTRACT One of the main forms of contamination of milk with mycotoxins is the consumption of fermented foods that are contaminated with mold, mainly Aspergillus spp, which produce toxins that can become a public health problem due to their thermal and chemical stability. The objective of the present work was to detect aflatoxin M1 concentrations in cows' milk samples in cooling tanks in four municipalities of the department of Boyacá for one year, determining the variations according to the season. A longitudinal, quantitative descriptive study was carried out, four cooling tanks from four different municipalities in the department were randomly selected, each tank, in each municipality, was sampled twice a month throughout the study period and processed using Charm methodology Ez Lite®, an ANDEVA was performed to determine the statistical differences between aflatoxin M1 concentrations for each quarter. Statistical differences were determined between each of the quarters of the study, finding a positivity percentage of 74.06% of the total positive samples in the summer quarters. 28.12% (108) of the samples taken throughout the study were positive, with toxin concentrations ranging between 0.5 and 2.0 μg/Kg of milk. Seasonal concentrations and variations of aflatoxin M1 in milk cooling tank samples were determined for the first time in the department of Boyacá, finding the highest concentrations and number of positive cases of aflatoxin M1 in the summer months.

The Occurrence and Risk Assessment of Exposure to Aflatoxin M₁ in Ultra-High Temperature and Pasteurized Milk in Hamadan Province of Iran

OBJECTIVES: Aflatoxins are a category of poisonous compounds found in most plants, milk and dairy products. The present research was carried out to detect the presence of aflatoxin M₁ (AFM₁) in samples of milk collected from Hamadan province, Iran. METHODS: Twenty five samples of ultra-high temperature (UHT) and 63 samples of pasteurized milk were collected and the amount of AFM₁ was measured by an Enzyme-Linked Immunosorbent Assay method. In addition, the estimated daily intake (EDI) and hazard index (HI) of AFM₁ was determined by the following equations:(EDI= mean concentration of AFM₁ × daily consumption of milk/body weight; HI=EDI/Tolerance Daily Intake). RESULTS: AFM₁ was detected in 21 (84%) UHT milk samples and in 55 (87.30%) pasteurized milk samples. Seven (28%) samples of UHT and 21 (33.33%) pasteurized milk samples had higher AFM₁ content than the limit allowed in the European Union and Iranian National Standard Limits (0.05 μg/kg). None of the samples exceeded the US Food and Drug Administration limit (0.5 μg/kg) for AFM₁. EDI and HI for AM₁ through milk were 0.107 ng/kg body weight/day, and 0.535, respectively. CONCLUSION: A significant percentage of milk produced by different factories in Iran (84% of UHT and 87.3% of pasteurized milk) was contaminated with AFM₁. Therefore, more control and monitoring of livestock feeding in dairy companies may help reduce milk contamination with AFM₁. As the HI value was lower than 1, it can be assumed that there was no risk of developing liver cancer due to milk consumption.

Aflatoxin M1 Reduction in Milk by a Novel Combination of Probiotic Bacterial and Yeast Strains

Milk and milk products are an important contribution to the human diet especially children. However, the presence of aflatoxins as AFM1 in milk and milk products are considered undesirables due to their health risks in consumer's body. For that reason, this study aimed to assess the ability of some microbial species on aflatoxin removal especially the AFM1 in the milk. The AFM1 residue was determined by HPLC after different incubation period (12 h, 24 h, 48 h and 72 h) of the probiotic bacteria and yeasts in PBS as a model for AFM1 determination. This study was performed during the period between 2015 and 2017. The combination of nonviable probiotic bacterial and yeast species (Lactobacillus Plantarum, Lactobacillus acidophilus, Bifidobacterium bifidum, Kluyveromyces lactis and Saccharomyces cerevisiae) succeeded to reduce AFM1 from 50 (ng/ml) during the incubation periods; 12 h, 24 h, 48 h and 72 h, into 9.72±1.31, 6.68±0.55, 5.70±0.33 and 4.56±0.15 ng ml-1, respectively. The highest AFM1 removal % was recorded as; 80.56%, 86.64%, 88.60% and 90.88% in the treated milk samples in a respective manner. Sensor evaluation was carried out using Yoghurt as a model in sample size 50 gm for each sample. Conclusion: This study concluded that the combination consisting of probiotic bacteria and yeasts could be used in reducing the concentration of the AFM1 in aflatoxin contaminated milk.

A comparative study of procedures for binding of aflatoxin M1 to Lactobacillus rhamnosus GG

ABSTRACT Several strains of lactic acid bacteria (LAB), frequently used in food fermentation and preservation, have been reported to bind different types of toxins in liquid media. This study was carried out to investigate the effect of different concentrations of Lactobacillus rhamnosus GG (ATCC 53103) to bind aflatoxin M1 (AFM1) in liquid media. AFM1 binding was tested following repetitive washes or filtration procedures in combination with additional treatments such as heating, pipetting, and centrifugation. The mixture of L. rhamnosus GG and AFM1 was incubated for 18 h at 37 °C and the binding efficiency was determined by quantifying the unbound AFM1 using HPLC. The stability of the complexes viable bacteria-AFM1 and heat treated bacteria-AFM1 was tested. Depending on the bacterial concentration and procedure used, the percentages of bound AFM1 by L. rhamnosus GG varied from as low as undetectable to as high as 63%. The highest reduction in the level of unbound AFM1 was recorded for the five washes procedure that involved heating and pipetting. Results also showed that binding was partially reversible and AFM1 was released after repeated washes. These findings highlight the effect of different treatments on the binding of AFM1 to L. rhamnosus GG in liquid matrix.

Assessment of aflatoxin M1 in breast milk of lactating mothers in Papua New Guinea

@#Aflatoxin M1 (AFM1) is a secondary metabolite in the breast milk of lactating mothers who consume foodstuffs infected by the fungi Aspergillus flavus and Aspergillus Parasiticus. The concentration of AFM1 in breast milk of lactating mothers is of major public health concern, because it can negatively affect the health of their babies. The major objective of this study was to assess the AFM1 concentration in the breast milk of lactating mothers in Papua New Guinea (PNG). This was a prospective cross-sectional study carried out between 2011 and 2015 in three of the four Regions in PNG: the National Capital District (NCD) in the Southern Region; Eastern Highlands (EHP) and Western Highlands (WHP) provinces in the Highlands Region; and East New Britain (ENB) and Manus provinces in the Islands Region. The Susu Mama, Well-Baby and Paediatric clinics in the General Hospitals in each of the selected provinces in the three regions were the primary sites for this study. A solid phase competitive Enzyme-Linked Immunosorbent Assay (ELISA 96 Microwell plates) was used for the quantification of AFM1 in breast milk from consented lactating mothers. A total of 874 lactating mothers and their babies participated in this study. The mean age of the mothers was 28.0 ±5.5 years. The age range of all the babies was 2 to 6 weeks. 76.1% (665/874) of all breast milk samples analyzed had detectable levels of AFM1. The concentration of AFM1 was above 10.00ppt in 89 (10.2%) of the 874 breast milk samples (which, according to the Australia / New Zealand / Austria safe cut-off limits for AFM1, makes them unsafe for consumption by the babies). The mean AFM1 concentration in the breast milk samples from lactating mothers in EHP (7.99ppt) was higher than that in the samples from the other 4 provinces in the present study. AFM1 concentration was above 10.00ppt in 14 (4.6%) of the 300 breast milk samples from NCD, in 62 (31.0%) of the 200 samples from EHP, in 10 (4.5%) of the 220 samples from ENB and in 3 (3.0%) of the 100 samples from WHP. In order to reduce the AFM1 concentrations in breast milk of lactating mothers, basic nutrition education, aggressive advocacy, social mobilization, awareness campaigns, including communication with all relevant target groups and the relevant policy makers are urgently required.

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

OBJECTIVES: Aflatoxin 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). METHODS: A total of 100 white cheese samples were randomly collected from supermarkets in Isfahan province and after extraction using dichloromethane were prepared for the ELISA. RESULTS: Of 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. CONCLUSION: Although 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.

Rapid detection of aflatoxin M1 in milk powder and milk based on up-converting phosphor technology / 军事医学

Objective To develop and evaluate an up-converting phosphor technology-based lateral flow assay ( UPT-LF) for detection of aflatoxin M1(AFM1) in milk powder and milk.Methods AFM1-UPT-LF was established with up-converting phosphor ( UCP) nano-particles as the bio-label of competitive mode based LF assay .Sensitivity, quantitative ability and precision were evaluated using simulated AFM 1-postive samples with serial standard concentrations .The qualita-tive and quantitative detection performance of AFM 1-UPT-LF was evaluated with reference to liquid chromatography-mass spectrography ( LC-MS) to detect samples of milk powder and milk simultaneously .Results AFM1-UPT-LF could conduct qualitative and quantitative detection without sample pretreatment within 20 min.The detection limit of AFM1-UPT-LF reached 0.1 μg/kg in milk powder and 0.3 μg/L in milk.There was good linearity ranging from 0.1 to 0.7 μg/kg and 0.3 to 0.7 μg/L for milk powder and milk, respectively.The sensitivity, specificity and receiver operating characteristic ( ROC) area under the curve ( AUC) of AFM1-UPT-LF for qualitative result could meet the need of national standards for AFM1 limit in dairy products.After statistical analysis, there was no significant difference (milk powder: t=0.66, P>0.05;milk:t=1.01, P>0.05) between AFM1-UPT-LF and LC-MS for quantitative detection .Conclusion The good qualitative and quantitative detection performance of AFM 1-UPT-LF for milk powder and milk makes possible on-site rapid detection of AFM1 in dairy products quantitatively .

Contaminants in milk and impact of heating: An assessment study.

Background: The major contaminants usually encountered in milk and milk products include pesticide residues, heavy metals, and aflatoxin M1 (AFM1). Primarily, milk get contaminated before milching, from the cattle feed, from sources/materials used during the processing of milk as well as improper handling of the milk during the pre- and postprocessing period. Objective: The purpose of this study was to evaluate the effect of household practices on milk contaminants. Materials and Methods: Samples of pasteurized as well as unpasteurized milk (Vendor's milk) were analyzed for AFM1, pesticide residues, and heavy metals. Simulating the household practices, the impact of boiling on these contaminants was assessed. Results: The contaminant Aflatoxin M1 (AFM1) was detected at a concentration ranging from 0.071-0.075 ppb in unpasteurized as well as pasteurized milk samples analyzed during the course of study. Moreover, boiling had no impact on the quantity of AFM1 present in the milk. Pesticides and heavy metal contents were found to be within acceptable limits in all the milk samples tested. Conclusion: Mycotoxins especially aflatoxins in cattle feed and their consequential presence in milk and milk products is a serious concern world over as they are reported carcinogens. These fungal toxins are resistant to high temperatures and may lead to various health hazards. Preventive steps must be taken at each stage to ensure good quality of milk and milk products free from these contaminants. Awareness programs and education for the dairy farmers and milk processors may be helpful in this regard.

Occurrence of aflatoxin M1 in parmesan cheese consumed in Minas Gerais, Brazil / Ocorrência de aflatoxina M1 em queijo Parmesão consumido em Minas Gerais, Brasil

Aflatoxina M1 (AFM1) pode ocorrer em leite e produtos de leite, resultante da ingestão de aflatoxina B1 presente em rações de gado leiteiro. Um total de 88 amostras de queijo Parmesão, comercializadas no estado de Minas Gerais, Brasil, no período de marçode 2004 a dezembro de 2004, foram analisadas para aflatoxina M1 por cromatografia líquida de alta eficiência (CLAE) com detector de fluorescência, precedida de purificação da amostra por coluna de imunoafinidade. AFM1 foi detectada em 40 das 88 amostras(46,4%). Entretanto, somente duas amostras entre estas estavam contaminadas em um nível acima do limite máximo permitido (250ng kg-1), aceito pela Comunidade Européia para AFM1. O Brasil não apresenta tolerância máxima permitida para AFM1 em queijo.

Aflatoxin M1 (AFM1) may occur in milk and milk products, resulting from the ingestion of aflatoxin B1 in feedstuffs by dairy cow. A total of 88 samples of Parmesan cheese marketed in Minas Gerais state, Brazil, from March 2004 to December 2004, were analyzed for AFM1 by high-performance liquid chromatography (HPLC) with a fluorescence detector following sample clean-up using immunoaffinity columns. AFM1 was detected in 40 of the 88 samples (46.4%). However, only two samples among these were contaminated at a level above the maximum permissible limit (250 ng kg-1) accepted by European Union for AFM1. No AFM1 maximum tolerance limit in cheese has been established in Brazil.

Gamma-irradiation effect on fumonisin B1 inactivation in corn flour and on aflatoxin M1 in fluid and powdered milk / Influência da irradiação gama na destruição de fumonisina B1 em farinha de milho e de aflatoxina M1 em leite fluido e em pó

Fumonisin B1 is a mycotoxin produced by Fusarium verticillioides and Fusarium proliferatum, and it is found chiefly in corn and corn-based products. Since its discovery fumonisin B1 has been associated with diseases in animals, such as leukoencephalomalacia in horses, and pulmonary edema in swine. On humans, the ingestion of foods with fumonisin B1 is associated with esophageal cancer. Aflatoxin M1 is the principal hydroxylated metabolite occurring in milk from animals which have consumed aflatoxin B1-contaminated feeds. It is also present in milk from nursing mothers who consumed foodstuffs with aflatoxin B1. In this study the effect of gamma-irradiation (60Co) was verified, in doses ranged from 0 to 20 kGy, in order to inactivate fumonisin B1 in corn flour and aflatoxin M1 in fluid and powdered milk. Fumonisin B1 was extracted from the samples with methanol:water (3:1). The extract was purified through immunoaffinity column followed by separation and quantification by means of high-performance liquid chromatography (HPLC) with o-phthaldialdehyde (OPA). For determining aflatoxin M1 the purification was done through immunoaffinity column followed by separation and quantification by means of HPLC with fluorescence detector. Gamma irradiation (60Co) at doses from 3 to 20 kGy reduced the fumonisin B1 content in a range of 11.2 % to 55.5 %. Gamma irradiation (60Co) at 20 kGy dose r

Fumonisina B1 é a micotoxina produzida por Fusarium verticillioides e Fusarium proliferatum e é encontrada principalmente em milho e produtos a base de milho. Desde sua descoberta a fumonisina B1 tem sido associada a doenças em animais, como leucoencefalomalácia em cavalos e edema pulmonar em suínos. Em humanos, o consumo de alimentos com fumonisina B1 tem sido associado com câncer esofágico. A aflatoxina M1 é o principal metabólito hidroxilado encontrado no leite de animais que consumiram rações contaminadas com aflatoxina B1, bem como no leite de lactantes que consumiram alimentos com esta substância. Neste estudo foi verificado o efeito da irradiação gama (60Co), em doses que variaram de 0 a 20 kGy, quanto à capacidade de inativar fumonisina B1 em farinha de milho e aflatoxina M1 em leite fluido e em pó. A fumonisina B1 foi extraída das amostras com metanol:água (8:2). O extrato foi purificado em coluna de imunoafinidade, seguido de separação e quantificação por meio de cromatografia líquida de alta eficiência (CLAE) com detector de fluorescência, após derivatização com ortoftaldialdeído. Para efetuar a determinação da aflatoxina M1, a amostra foi purificada em coluna de imunoafinidade e a separação e a quantificação por meio de CLAE com detector de fluorescência. Foi observada uma redução da concentração da fumonisina B1 na faixa de 11,2 % a 55,5% em doses de 3 a 20 kGy de