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PubMed2006

Aflatoxin M1 effects on Xenopus laevis development.

BACKGROUND: The principal Aflatoxin B(1) (AFB(1)) hydroxylated metabolite excreted in milk is Aflatoxin M(1) (AFM(1)) classified in group 2B by the International Agency for Research on Cancer (IARC). Human exposure to AFM(1) is due to the consumption of contaminated dairy products and partly to endogenous production through AFB(1) liver metabolism. METHODS: Since no data are available on AFM(1) embryotoxicity, its lethal and teratogenic potential was investigated using the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). Stage-8 blastulae were exposed to AFM(1) at 1, 4, 16, 64, and 256 microg/L concentrations until stage 47, free-swimming larva. RESULTS: A slight increase of mortality and malformed larva percents was found in AFM(1)-exposed groups but these differences were not statistically significant in comparison with the controls. CONCLUSIONS: Therefore, AFM(1) is a non-embryotoxic compound when evaluated with a FETAX model at concentrations under the conditions tested. However, AFM(1) merits further studies using mammals as experimental models to identify a possible risk during human pregnancy.

Aflatoxin M1↗
PubMed1996

Food surveillance in the Basque Country (Spain). II. Estimation of the dietary intake of organochlorine pesticides, heavy metals, arsenic, aflatoxin M1, iron and zinc through the Total Diet Study, 1990/91.

Total diet samples purchased at monthly intervals between March 1990 and December 1991 were analysed for different contaminants and nutrients. Each total diet sample included 91 food items which were combined after preparation and/or cooking into 16 groups of similar foods. The 'market basket' was based on a food survey which referred to the adult population (25-60 years) carried out in the Basque Country between 1988 and 1990. The dietary intakes (micrograms/day) of lead (43), cadmium (11), mercury (18), arsenic (291), hexachlorobenzene (HCB) (0.2), alpha-hexachlorocyclohexane (alpha-HCH) ( < 0.1), beta-hexachlorocyclohexane (beta-HCH) (0.1), gamma-hexachlorocyclohexane (gamma-HCH) (2.9), dichlorodiphenyltrichloroethane (DDT) (0.3), dichlorodiphenyldichloroethene (DDE) (0.9), dichlorodiphenyldichloroethane (DDD)(0.2), dieldrin (0.5), heptachlor epoxide ( < 0.1), alpha-endosulphan (0.1) and beta-endosulphan (0.1) were all well below the respective Acceptable Daily Intakes or Provisional Tolerable Weekly Intakes. However, arsenic intake was much higher than that estimated in other countries and gamma-HCH was detected in anomalously high levels in the bread group. Dietary intakes of delta-hexachlorocyclohexane (delta-HCH), aldrin, endrin, heptachlor and methoxychlor were not calculated because no residues were detected in any of the samples. Aflatoxin M1 intake was not estimated owing to the low levels detected. Finally, zinc intakes (11.6 mg/day) were below the recommended dietary allowances for Spain and the same was true for iron (11.3 mg/day), but only for females.

Adult↗
PubMed2005

Development and validation of an ultrasensitive chemiluminescent enzyme immunoassay for aflatoxin M1 in milk.

A fast and ultrasensitive chemiluminescent enzyme immunoassay for aflatoxin M(1) in milk samples has been developed and validated. The method is an indirect competitive type format involving the immobilization of an aflatoxin M(1)-bovine serum albumin conjugate on 384 well black polystyrene microtiter plates and the use of a secondary antibody labeled with horseradish peroxidase detected with a luminol-based substrate. Aflatoxin M(1) standard solutions were prepared in milk-based buffer, and milk samples were analyzed without any cleanup procedure. The limit of quantification was 1 ppt, the coefficient of variation was below 9% for both intra- and interassay precision, and the recovery ranged from 96 to 122%. The method is specific, and other aflatoxins do not significantly cross-react with the antibody. Twenty-four milk samples were analyzed, and a good correlation was observed (y = 0.98x + 1.71, r(2) = 0.98, n = 24) when the data were compared with a reference high-performance liquid chromatography method with a fluorescent detector. The developed method is suitable for an accurate, sensitive, and high-throughput screening of aflatoxin M(1) in milk samples with a reduction of costs and increased detectability, as compared with previously developed immunoassays.

Aflatoxin M1↗
PubMed2006

Aflatoxin M1 and cyclopiazonic acid in fluid milk traded in São Paulo, Brazil.

Aflatoxin M(1) (AFM(1)) and cyclopiazonic acid (CPA) were determined in 48 samples of grade A-C and ultra-high-temperature (UHT)-treated fluid commercial milk from São Paulo, Brazil, from July to December 2004. AFM(1) and CPA determinations were carried out by HPLC. Results showed 37 (77.1%) positive samples for AFM(1) at levels of 0.011-0.251 microg l(-1) milk, which were below the tolerance limit for AFM(1) in milk as adopted by Brazilian regulations. Mean levels of AFM(1) in grades A-C and UHT milk were 0.060+/-0.080, 0.056+/-0.063, 0.064+/-0.041 and 0.075+/-0.048microg l(-1), respectively. CPA was detected in two samples (4.2%) of grade A milk at levels of 6.4 and 9.7 microg l(-1). This is the first report of the occurrence of CPA in milk in Brazil. It was concluded that the incidence of AFM(1) in milk traded in São Paulo is low, but the simultaneous occurrence of CPA, even at low levels, warrants concern about AFM(1) in milk and milk products.

Aflatoxin M1↗
PubMed2006

Distribution and stability of Aflatoxin M1 during processing and ripening of traditional white pickled cheese.

The distribution of aflatoxin M(1) (AFM(1)) has been studied between curd, whey, cheese and pickle samples of Turkish white pickled cheese produced according to traditional techniques and its stability studied during the ripening period. Cheeses were produced in three cheese-making trials using raw milk that was artificially contaminated with AFM(1) at the levels of 50, 250 and 750 ng/l and allowed to ripen for three months. AFM(1) determinations were carried out at intervals by LC with fluorescence detection after immunoaffinity column clean-up. During the syneresis of the cheese a proportionately high concentration of AFM(1) remained in curd and for each trial the level was 3.6, 3.8 and 4.0 times higher than levels in milk. At the end of the ripening, the distribution of AFM(1) for cheese/whey + brine samples was 0.9, 1.0 and 1.3 for first, second and third spiking respectively indicating that nearly half of the AFM(1) remained in cheese. It has been found that only 2-4% of the initial spiking of AFM(1) transferred into the brine solution. During the ripening period AFM(1) levels remained constant suggesting that AFM(1) was quite stable during manufacturing and ripening.

Aflatoxin M1↗
PubMed2006

Determinants of aflatoxin M1 in breast milk in a selected group of Egyptian mothers.

In Egypt, there is a paucity of biomarker data on aflatoxin (AF) exposure. The study assessed the level and frequency of breast milk AFM1 as a biomarker of maternal exposure. Breast milk samples were collected from a selected group of 388 Egyptian lactating mothers of children attending the New El-Qalyub Hospital, Qalyubiyah governorate, Egypt, during May-September 2003. Following aflatoxin extraction, AFM1 levels were assessed by high-performance liquid chromatography (HPLC) with fluorescence detection. Approximately 36% of mothers tested positive for AFM1 (median 13.5 pg ml-1, interquartile range (IQR) 10.27-21.43). Non-working status (p = 0.018, odds ratio (OR) = 2.87), obesity (p = 0.004, OR = 3.01), high corn oil consumption (p = 0.002, OR = 2.21), number of children (>1) (p = 0.025, OR = 1.99), and early lactation stage (<1 month) (p = 0.028 OR = 3.57), contributed to the occurrence of AF in breast milk. AFM1 contamination of breast milk was frequent, albeit at moderate levels. Growth and development of the infant is rapid and thus it is possible that AF exposure through breast milk has a significant health effect.

Adult↗
PubMed1988

An epidemiological investigation associating aflatoxin M1 with milk production in dairy cattle.

An attempt was made to utilize the analysis for aflatoxin M2 in milk, instead of aflatoxin B1 in feed for a pilot epidemiological study addressing the association of M1 levels and various production parameters. Four dairy farms, representing above average to below average management practices, were identified with 10 animals being randomly selected from each farm for this study. Milk samples and production data were evaluated from these animals in order to gain insight into the possible effects of low level mycotoxin ingestion on production and to test whether or not this approach offered potential for future epidemiological studies. Feed samples were basically negative for the presence of aflatoxin B1, yet milk analysis revealed the presence of M1 in a large percentage of the animals. Further, the M1 levels appeared to be associated with a decrease in daily milk production. This study indicates that employing M1 analysis in milk for epidemiological investigations of low level mycotoxin ingestion effects, instead of B1 feed analysis, is feasible and offers a potential for more definitive studies in this area.

Aflatoxin M1↗
PubMed1997

Use of an improved method for analysis of urinary aflatoxin M1 in a survey of mainland China and Taiwan.

An improved monoclonal antibody immunoaffinity chromatography/high-pressure liquid chromatography/ fluorescence detection method was developed to measure aflatoxin (AF) exposure by quantifying AFM1 in human and rat urine samples. Analysis of different amounts of various AF metabolites showed that the immunoaffinity resin was highly selective for aflatoxin B1 (AFB1), AFB2, and AFM1. Recovery of added AFs increased with the amount of immunoaffinity resin and was virtually complete within the range of 0.01-10 ng of AFM1 by using 7 ml of resin. The detection limit of this method is 0.5 pg/ml urine. Rats dosed with tritiated AFB1 excreted in their urine tritiated AFM1, among other AF metabolites, as indicated by chemical derivative confirmation and cochromatography with authentic AFM1 and agreement of radioactivity and fluorescence quantitation. A linear dose-response relationship was found over the range of 0.05-50 micrograms/kg of body weight/day. Two humans dosed with 1.0 microgram of pure AFB1 excreted 6-7% of the dose as urinary AFM1 over 5-7 days. Pooled urine samples from 30 men from each of 69 rural counties in mainland China and 16 survey areas in Taiwan, with two villages per county or area, were analyzed with this improved method (170 villages total). The correlation coefficient of urinary excretion of AFM1 compared between villages within all 85 survey areas was 0.50 (P < 0.001). Sixty-five % of the samples contained detectable concentrations of AFM1 with an average excretion of 3.1 ng/12 h. Assuming an excretion rate of 2-6%, this AFM1 excretion corresponds to a very low average daily AF consumption of 0.1-0.3 microgram/day (possible range, 0-11 micrograms/day). Patterns of urinary excretion of AFM1 were similar in mainland China and Taiwan.

Adult↗
PubMed2003

Aflatoxin M1 in breast-milk of UAE women.

A study was undertaken to determine whether breast-milk of mothers from the United Arab Emirates (UAE) contained aflatoxins. One hundred and forty lactating mothers, 55 who had delivered premature infants (<2500 g, </=37 wk gestational age) and 85 who had full-term infants, agreed to participate in the study. Breast-milk was collected during regular feeding of infants in the special care baby unit and postnatal wards using an electric breast pump and ten ml of milk was siphoned off into a zinc-free plastic container for analysis. Aflatoxin M(1) concentration in the breast-milk samples was measured by HPLC. Samples were collected between January 1999 and December 2000. Approximately 66% of the mothers were expatriates and 34% were UAE nationals. Babies' weight, postnatal age, sex, birthweight and gestational age, and mothers' nationality, age and parity were recorded. Overall, 92% of the breast-milk samples contained aflatoxin M(1). Both univariate and multivariate logistic regression analysis failed to show significant correlation between aflatoxin M(1) and gestational age, postnatal age, gender, nationality and clinical condition. Levels of lactose, protein and lipids did not correlate significantly with levels of aflatoxin M(1). The public should be educated about storing food and the hazards of aflatoxin ingestion to reduce the unacceptably frequent presence of aflatoxin in breast-milk.

Aflatoxin M1↗
PubMed2006

Survey of the occurrence of Aflatoxin M1 in ovine milk by HPLC and its confirmation by MS.

During the period of October-July 2000, 240 samples of dairy ewes milk, obtained from farms of Enna (Sicily, Italy), were checked for Aflatoxin M(1) (AFM(1)) by HPLC using a fluorimetric detector. The limit of detection and the limit of quantification were 250 ng/L for AFM(1). All the positive milk samples for AFM(1) were confirmed by LC-MS. AFM(1) was detected in 81% of milk samples, ranging from 2 to 108 ng/L. Three samples were over the legal limits (50 ng/L). Mean contamination of samples obtained from stabulated ewes was higher than that from grazing ewes (35.27 vs. 12.47 ng/L). Furthermore, samples collected in the period September-October showed higher contamination than samples collected during the other months (42.68 vs. 10.55 ng/L). Both differences are related to the administration of compound feed. Based on current toxicological knowledge we concluded that the AFM(1) contamination levels recorded in ewe milk did not present a serious human health hazard. However, as ewe milk is exclusively used to produce cheese due to its higher protein content, and also considering the preferential binding of AFM(1) to casein during coagulation of milk, a potentially high concentration effect could occur, thus the surveillance of contamination levels should be more continuous and widespread.

Aflatoxin M1↗
PubMed2004

Joint IDF-IUPAC-IAEA(FAO) interlaboratory validation for determining aflatoxin M1 in milk by using immunoaffinity clean-up before thin-layer chromatography.

A collaborative study was conducted under the auspices of the International Dairy Federation (IDF), the International Union of Pure and Applied Chemistry (IUPAC) and the International Atomic Energy Agency (IAEA), a collaborative Food and Agricultural Organization (FAO) body fully to validate a method combining immunoaffinity clean-up to thin-layer chromatography for the determination of aflatoxin M(1) in milk. Work was done in order to afford those laboratories not equipped with high-performance liquid chromatography, mainly from developing countries, with a simplified but fully validated method as an alternative to the European validated immunoaffinity-high performance liquid chromatography method published as an EN ISO Standard 14501, February 1999. The validation study was carried out on samples of aflatoxin M(1)-contaminated milk and milk powder at levels close to the tolerable level of 0.5 microg l(-1) as recommended by the Codex Alimentarius and to the regulatory level of 0.05 microg l(-1) as laid down by the European Commission. Fourteen laboratories representing 11 countries participated in the trial. The relative standard deviations for repeatability and reproducibility based on raw data were in the range 27-48 and 35-54%, respectively. The recovery rate varied from 32 to 120%. The mishandling of two crucial steps of the protocol such as matrix sample reconstitution and extract evaporation could explain the wide variation of the recovery rate. For this reason, data were then corrected for recovery. Consequently, the relative standard deviations for repeatability and reproducibility were recalculated after correction for recovery and were in the range 26-54 and 34-53%, respectively. The method will be published as a standard ISO/DIS 14674--IDF 190.

Aflatoxin M1↗
PubMed2005

Aflatoxin occurrence in foodstuff supplied to dairy cattle and aflatoxin M1 in raw milk in the North of Paraná state.

Moulds occur in a great variety of foods, including the concentrated and roughage destined for animal feeding, and can produce mycotoxins under certain conditions. The ingestion of mycotoxin AFB(1) by dairy cattle leads to the biotransformation of that substance, which is eliminated via milk as AFM(1), and also causes damage to human health. The present study aimed to analyze the presence of aflatoxins in foodstuff destined for dairy cattle and in the milk produced by these animals. The contamination of the foodstuff by aflatoxins happened mainly in the feeds and the silages did not present contamination by aflatoxins. Out of the 42 samples of milk analyzed, 10 (24%) were contaminated by AFM(1), and 3 (7%) were above the 0.5 microg/l limit.

Aflatoxin B1↗
PubMed2001

Distribution of aflatoxin M1 in cheese obtained from milk artificially contaminated.

Small-scale manufacture of cheese using artificially AFM1 contaminated milk as raw material to study the distribution of such toxin both in whey and in cheese, was carried out. Whole milk with undetectable levels of AFM1 was used. The toxin was added in concentration that varied from 1.7 to 2.0 microg/l of milk. After the home-made production of cheese, the concentration of AFM1 was determined both in whey and in cheese, using the enzymatic immunoassay technique. The greatest proportion, 60%, was detected in whey while 40% AFM1 remained in cheese.

Aflatoxin M1↗