Cytotoxicity of gamma irradiated aflatoxin B1 and ochratoxin A

Toxicity of gamma irradiated mycotoxins aflatoxin B₁ (AFB₁) and ochratoxin A (OTA) was investigated in vitro. AFB₁ and OTA stock solutions (50?mM, in methanol) were gamma irradiated (5 and 10 kGy) and non-irradiated and irradiated mycotoxins solutions were tested for cytotoxicity on Pk15, HepG2 and SH-SY5Y cell lines (MTT assay, 1–500?μM concentration range; 24?h exposure). Degradation of mycotoxin molecules was examined by liquid chromatography tandem mass spectrometry (HPLC-MS/MS). AFB₁ and OTA radiolytic products were less toxic than the parent mycotoxins to all of the tested cell lines. Gamma irradiation even at 5 kGy had effect on AFB₁ and OTA molecules however, this effect was dependent on chemical structure of mycotoxin. Since gamma irradiation at low dose reduced initial level of both mycotoxins, and gamma irradiated mycotoxins had lower toxicity in comparison to non-irradiated mycotoxins, it can be concluded that gamma irradiation could be used as decontamination method.     

Application of a minicolumn detection method for screening spices for aflatoxin

The minicolumn of Holaday and Lansden was modified by increasing the height of neutral alumina and including a layer of anhydrous sodium sulphate. Using this procedure, aflatoxin was detected in 18 of 125 samples of black pepper, red pepper, ginger and turmeric. A few samples of each spice contained aflatoxin, although red pepper and turmeric showed the highest incidence. Aflatoxin B1 quantities ranged from 10 ug/kg to 120 ug/kg when estimated quantitatively by visual comparison with standards on thin-layer chromatography plates.     

Effects of probiotic bacteria on the bioaccessibility of aflatoxin B1 and ochratoxin A using an in vitro digestion model under fed conditions

In the present study, we aimed at determining the release of aflatoxin B₁ (AFB₁) and ochratoxin A (OTA) from different food products in the gastro-intestinal tract in the absence and presence of probiotics, a possible adsorbent. The average bioaccessibility of AFB₁ and OTA without probiotics was about 90%, and 30%, respectively, depending on several factors, such as food product, contamination level, compound and type of contamination (spiked versus naturally contaminated). The six probiotic bacteria showed varying binding capacity to AFB₁ and OTA depending on the bacterial strain, toxin studied, type of food and contamination level. A reduction to a maximum of 37% and 73% as observed for the bioaccessibility of AFB₁ and OTA in the presence of probiotic bacteria, respectively. This is the first report on the effect of probiotic bacteria on reducing the fraction of mycotoxins available for absorption in the gastrointestinal tract from different food products.     

Aflatoxin decomposition in various soils

The persistence of aflatoxin in the soil environment could potentially result in a number of adverse environmental consequences. To determine the persistence of aflatoxin in soil, 14C-labeled aflatoxin B1, was added to silt loam, sandy loam, and silty clay loam soils and the subsequent release of 14CO2 was determined. After 120 days of incubation, 8.1% of the original aflatoxin added to the silt loam soil was released as CO2. Aflatoxin decomposition in the sandy loam soil proceeded more quickly than the other two soils for the first 20 days of incubation. After this time, the decomposition rate declined and by the end of the study, 4.9% of the aflatoxin was released as CO2. Aflatoxin decomposition proceeded most slowly in the silty clay loam soil. Only 1.4% of aflatoxin added to the soil was released as CO2 after 120 days incubation. To determine whether aflatoxin was bound to the silty clay loam soil, aflatoxin B1 was added to this soil and incubated for 20 days. The soil was periodically extracted and the aflatoxin species present were determined using thin layer chromatographic (TLC) procedures. After one day of incubation, the degradation products, aflatoxins B2 and G2, were observed. It was also found that much of the aflatoxin extracted from the soil was not mobile with the TLC solvent system used. This indicated that a conjugate may have formed and thus may be responsible for the lack of aflatoxin decomposition.     

Aflatoxin M1 contamination of milk and ice cream in Abeokuta and Odeda local governments of Ogun State, Nigeria

A survey was undertaken to determine the aflatoxin M(1) contamination of milk and some locally produced dairy products in Abeokuta and Odeda local governments of Ogun State, Nigeria. Samples of human and cow milk, yoghurt, "wara", ice cream and "nono" were collected randomly within the local governments and analysed for aflatoxin M(1) using the two-dimensional TLC. Aflatoxin M(1) contamination in the range of 2.04-4.00 microg l(-1) was noticed only in milk and ice cream. In particular, samples of human milk, cow milk and ice cream recorded high scores of 4.0 microg l(-1), 2.04 microg l(-1) and 2.23 microg l(-1), respectively in Abeokuta local governments and a score of 4.0 microg l(-1) for cow milk in Odeda local government. This indicates a high level contamination in the local governments since the weighted mean concentration of aflatoxin M1 in milk for African diet is 0.002 microg l(-1). Therefore the concentration of AFB1 in feeds which is transformed to AFM1 in milk should be reduced by good manufacturing and good storage practices. Furthermore, there is need for stringent quality control during processing and distribution of these products.     

Application of a LC–MS/MS method for multi-mycotoxin analysis in infant formula and milk-based products for young children commercialized in Southern Brazil

ABSTRACT

An analytical method based on liquid chromatography–tandem mass spectrometry (LC–MS/MS) was validated and applied for the analysis of aflatoxin M₁ (AFM₁), ochratoxin A (OTA) and deoxynivalenol (DON) in infant formula and milk-based products for young children commercialized in Brazil. A total of 38 samples were evaluated, including 12 infant formula, 14 follow-on formula and 12 samples of milk-based products. AFM₁ was detected in 12 (32%) samples, and seven (18%) samples contained AFM₁ levels above the method limit of quantification in a concentration range between 0.013 and 0.067 ng mL^?1 (0.026 ± 0.019). Two samples of milk-based products exceeded the maximum level (ML) fixed by the European Union for AFM₁ in baby foods, however, all samples were in agreement with the levels established by the Brazilian regulation. OTA and DON were not detected in any of the analyzed samples.     

The aflatoxin contamination of some pepper types sold in Turkey

In the present study, 44 red-scaled pepper and 26 red powder pepper samples obtained from various spice retailers in Erzurum, and 20 isot (red pepper produced in Sanliurfa) were investigated for aflatoxin contamination using thin layer chromatography. Aflatoxin (B + G) was found in 8 red pepper samples (18.2%), and 3 red powder peppers (10.7%), and 1 isot sample (5%). The aflatoxin amount ranged from 1.1 to 97.5 ppb in all samples investigated. The highest amount of aflatoxin was found in red-scaled pepper. Species of Aspergillus, Penicillium and Rhizopus were generally contaminants of all samples.     

Effects of the aflatoxins on ATPase activities in mouse and rat liver.

The effects of the aflatoxins on ATPase activities in mouse and rat tissues were investigated in vitro. The hepatic oligomycin-sensitive (O.S.) Mg++ ATPase was inhibited significantly. The order of inhibition was G1 greater than B1 greater than G2 greater than B2. Mouse O.S. Mg++ ATPase was more sensitive than the corresponding rat enzyme. The oligomycin-insensitive (O.I.) Mg++ ATPase activities in rat and mouse liver were not altered. Although aflatoxins G1 and B1 were more potent inhibitors of hepatic O.S. Mg++ ATPase, no concentration-response was observed, whereas aflatoxins G2 and B2 inhibited enzyme activity in a concentration-dependent manner. Spectral analysis of aflatoxin G1 solutions suggested that solubility was not related to the observed effects. In addition, the effects of aflatoxin B1 and G1 on mouse brain microsomal Na+-K+ ATPase were examined. Although aflatoxin B1 was more potent that G1, both mycotoxins significantly inhibited enzyme activity in a concentration-dependent fashion.     

Aflatoxin decomposition in various soils

Abstract

The persistence of aflatoxin in the soil environment could potentially result in a number of adverse environmental consequences. To determine the persistence of aflatoxin in soil, ¹⁴C‐labeled aflatoxin B₁, was added to silt loam, sandy loam, and silty clay loam soils and the subsequent release of ¹⁴CO₂ was determined. After 120 days of incubation, 8.1% of the original aflatoxin added to the silt loam soil was released as CO₂ ? Aflatoxin decomposition in the sandy loam soil proceeded more quickly than the other two soils for the first 20 days of incubation. After this time, the decomposition rate declined and by the end of the study, 4.9% of the aflatoxin was released as CO_2. Aflatoxin decomposition proceeded most slowly in the silty clay loam soil. Only 1.4% of aflatoxin added to the soil was released as CO₂ after 120 days incubation. To determine whether aflatoxin was bound to the silty clay loam soil, aflatoxin B₁ was added to this soil and incubated for 20 days. The soil was periodically extracted and the aflatoxin species present were determined using thin layer chromatographic (TLC) procedures. After one day of incubation, the degradation products, aflatoxins B₂ and G₂, were observed. It was also found that much of the aflatoxin extracted from the soil was not mobile with the TLC solvent system used. This indicated that a conjugate may have formed and thus may be responsible for the lack of aflatoxin decomposition.     

The incidence of fungi and mycotoxins in South Africa wheat and wheat-based products

This investigation was undertaken to survey the fungal and mycotoxin contamination of South African wheat ranging from that growing in the field to processed wheat products. Samples of wheat were taken from various growing areas in South Africa and screened for fungi and mycotoxins, using a range of methodologies, including chromatography, immunoaffinity/fluorimetry, and cytotoxicity testing. Similar samples were taken from supermarkets and retail outlets in South Africa and analyzed in a similar manner. The result showed that a range of fungi and mycotoxins could be detected in wheat in all these sample types. The major fungal contaminants were Fusarium spp. and their attendant mycotoxins, in particular deoxynivalenol, which is in keeping with the observations made in the rest of the world. An interesting observation was that samples of wheat taken from the field with heavy Fusarium contamination were contaminated with fumonisin B1, which is not normally associated with this crop. Of more concern were the low but persistent levels of mycotoxins and fungi in wheat-based products sold directly to the public.     

Occurrence of aflatoxin M1 in human milk samples in Vojvodina,Serbia: Estimation of average daily intake by babies

The objectives of the study were to determine the aflatoxin M1 content in human milk samples in Vojvodina, Serbia, and to assess the risk of infants' exposure to aflatoxins food contamination. The growth of Aspergillus flavus and production of aflatoxin B1 in corn samples resulted in higher concentrations of AFM1 in milk and dairy products in 2013, indicating higher concentrations of AFM1 in human milk samples in 2013 and 2014 in Serbia. A total number of 60 samples of human milk (colostrum and breast milk collected 4–8 months after delivery) were analyzed for the presence of AFM1 using the Enzyme Linked Immunosorbent Assay method. The estimated daily intake of AFM1 through breastfeeding was calculated for the colostrum samples using an average intake of 60 mL/kg body weight (b.w.)/day on the third day of lactation. All breast milk collected 4–8 months after delivery and 36.4% of colostrum samples were contaminated with AFM1. The greatest percentage of contaminated colostrum (85%) and all samples of breast milk collected 4–8 months after delivery had AFM1 concentration above maximum allowable concentration according to the Regulation on health safety of dietetic products. The mean daily intake of AFM1 in colostrum was 2.65 ng/kg bw/day. Results of our study indicate the high risk of infants' exposure, who are at the early stage of development and vulnerable to toxic contaminants.     

Inhibition of pancreatic carboxypeptidase A: A possible mechanism of interaction between penicillic acid and ochratoxin A

Abstract

Penicillic acid and ochratoxin A are environmentally important toxic fungal metabolites (mycotoxins) that are synergistic in combination. The effects of penicillic acid on the pancreatic enzyme, :arboxypeptidase A were investigated in vitro and in vivo. A broad range of inhibition in vitro of the enzyme by PA was demonstrated with a half‐maximal inhibitory concentration equal to 1.1 x 10^‐4M PA. Inhibition of carboxypeptidase A was time and temperature dependent, and resulted in decreased conversion of parent ochratoxin A to the non‐toxic metabolite, ochratoxin alpha. Studies in vivo demonstrated a penicillic acid‐dependent inhibition of pancreatic carboxypeptidase A activity in the mouse and the chicken following multiple oral exposure. It is postulated that the mode of toxic interaction of the two mycotoxins may be due, in part, to impaired detoxification of ochratoxin A through peni‐cillic acid depletion of carboxypeptidase A activity.     

The mediating role of coagulation function on the association of prenatal exposure to aflatoxin B1 and postpartum hemorrhage in Guangxi,China

Environmental Science and Pollution Research - Pregnant women are vulnerable to certain environmental agents, one of which is aflatoxin. As one of the most popular aflatoxins, Aflatoxin B1 (AFB1)...     

The Incidence of Fungi and Mycotoxins in South Africa Wheat and Wheat-Based Products

This investigation was undertaken to survey the fungal and mycotoxin contamination of South African wheat ranging from that growing in the field to processed wheat products. Samples of wheat were taken from various growing areas in South Africa and screened for fungi and mycotoxins, using a range of methodologies, including chromatography, immunoaffinity/fluorimetry, and cytotoxicity testing. Similar samples were taken from supermarkets and retail outlets in South Africa and analyzed in a similar manner. The result showed that a range of fungi and mycotoxins could be detected in wheat in all these sample types. The major fungal contaminants were Fusarium spp. and their attendant mycotoxins, in particular deoxynivalenol, which is in keeping with the observations made in the rest of the world. An interesting observation was that samples of wheat taken from the field with heavy Fusarium contamination were contaminated with fumonisin B₁, which is not normally associated with this crop. Of more concern were the low but persistent levels of mycotoxins and fungi in wheat-based products sold directly to the public.     

Monitoring of bisphenol A diglycidyl ether (BADGE) and some derivatives in fish products in the Turkey market

The exposure to bisphenols and their derivatives was assessed in 33 fish products sold in Turkey using high-performance liquid chromatography-tandem mass spectrometry (LC–MS/MS). BADGE was determined in only four samples at concentrations ranging between 0.06 and 0.22 mg/kg. As the most abundant bisphenol groups, BADGE-hydrolyzed products such as BADGE·H₂O and BADGE·2H₂O were present in nine and fourteen samples in the range between 0.06–0.16 and 0.06–0.72 mg/kg, respectively. The total concentration of BADGE and hydrolyzed products was below the specific migration limit (SML) value of 9 mg/kg food, which in the European Union stated as tolerable. Chlorinated derivatives of BADGE were detected in fewer samples compared with hydrolyzed ones. BADGE·H₂O·HCl was the predominant migrant among chlorinated derivatives and was present in seven samples in a range between 0.02 and 0.06 mg/kg. All other samples contained less than or equal to 0.03 mg/kg of BADGE·HCl and BADGE·2HCl. The sum of these derivatives was lower than the SML value (1 mg/kg) of BADGE chlorohydrins legislated by the European Union. Besides these migrants, the analyzed samples did not contain any BFDGE and 3R-NOGE, which are prohibited in manufacturing food contact materials.

     

Results of the monitoring program of pesticide residues in organic food of plant origin in Lombardy (Italy)

Organic agriculture, with its restrictions on the use of synthetic chemical inputs, seems to offer a low-residue alternative to conventional methods. In Europe, the Council Regulation n. 2092/91/EEC regulates the production and trade of organic products and foodstuffs; national and regional legislation in Italy gives specific guidance on the surveillance of organic agriculture. However, monitoring of specific chemical residues in organic foodstuffs is part of the regular controls on food, aiming to safeguard consumer's health. Monitoring programs are coordinated at the national level by the Ministry of Health and at local level by Regional authorities. In Lombardy, in accordance with the provisions of the General Directorate of Health of the Region and under the supervision of the 15 Local Health Units, a monitoring program of pesticide residues in food of plant origin is undertaken every year. The International Centre for Pesticides and Health Risk Prevention (ICPS), on behalf of the General Directorate of Health of the Region of Lombardy, has been collecting and elaborating the data resulting from the analysis of food samples, carried out by the local laboratories. During the period 2002–2005, a total of 3508 samples food of plant origin were analyzed for pesticide residues, among which were 266 samples of organic farming products. Commodities were classified into groups (citrus fruits, legumes, vegetables, potatoes, processed products, cereals, and fruit other than citrus) and the outcomes of the analyses were reported by year, origin of the sample and presence/absence of pesticide residues. Results showed that the vast majority of organic farming products were in conformity with the relevant legislation and did not contain detectable pesticide residues. A limited amount of samples had residues at concentration below the Maximum Residue Limit (MRL). Only in one sample the residue level was above the MRL, however it did not pose a concern for public health, as demonstrated by the outcomes of dietary risk assessment. Organic fruits and vegetables can be expected to contain fewer agrochemical residues than conventionally grown alternatives. There is a widespread belief that organic agriculture products are safer and healthier than conventional foods. It is difficult to come to conclusions, but what should be made clear to the consumer is that “organic” does not automatically equal “safe”. In the absence of adequate comparative data, additional studies in this area of research are required.     

The incidence of fungi and mycotoxins in South African barley and barley products

Barley is grown as a crop in South Africa, mainly for use in the barley beer (lager) industry, particularly in the production of barley malt. This investigation was done to find out what fungal infection and mycotoxin contamination this barley and the malt contained. The survey, done in 2005, not only covered barley and malt but also the final product in the form of several brands of beer purchased from retail outlets. Analysis was done using a range of methodologies including chromatography, immunoaffinity/fluorimetry and cytotoxicity testing. The results show that barley, which also includes barley sold directly to the public, and malt, contain various fungi, sometimes at high incidence and also a range of mycotoxins which persisted through, although at low levels, to beer. The malt showed a different pattern of micro flora, as compared to the barley, which indicates infection during the malting process, which is not uncommon. Interestingly, fumonisin B1 was found in some of the samples, as well as the beer, although in the latter case these were at very low levels. Recent studies have shown that the immunoaffinity/fluorimetry method can give false positives for the fumonisins, so these results were confirmed by high performance liquid chromatography. As is the case with other cereals grown in South Africa, there is the concern that the public is exposed to mycotoxins on a regular basis through the consumption of products made from them.     

Results of the monitoring program of pesticide residues in organic food of plant origin in Lombardy (Italy)

Organic agriculture, with its restrictions on the use of synthetic chemical inputs, seems to offer a low-residue alternative to conventional methods. In Europe, the Council Regulation n. 2092/91/EEC regulates the production and trade of organic products and foodstuffs; national and regional legislation in Italy gives specific guidance on the surveillance of organic agriculture. However, monitoring of specific chemical residues in organic foodstuffs is part of the regular controls on food, aiming to safeguard consumer's health. Monitoring programs are coordinated at the national level by the Ministry of Health and at local level by Regional authorities. In Lombardy, in accordance with the provisions of the General Directorate of Health of the Region and under the supervision of the 15 Local Health Units, a monitoring program of pesticide residues in food of plant origin is undertaken every year. The International Centre for Pesticides and Health Risk Prevention (ICPS), on behalf of the General Directorate of Health of the Region of Lombardy, has been collecting and elaborating the data resulting from the analysis of food samples, carried out by the local laboratories. During the period 2002-2005, a total of 3508 samples food of plant origin were analyzed for pesticide residues, among which were 266 samples of organic farming products. Commodities were classified into groups (citrus fruits, legumes, vegetables, potatoes, processed products, cereals, and fruit other than citrus) and the outcomes of the analyses were reported by year, origin of the sample and presence/absence of pesticide residues. Results showed that the vast majority of organic farming products were in conformity with the relevant legislation and did not contain detectable pesticide residues. A limited amount of samples had residues at concentration below the Maximum Residue Limit (MRL). Only in one sample the residue level was above the MRL, however it did not pose a concern for public health, as demonstrated by the outcomes of dietary risk assessment. Organic fruits and vegetables can be expected to contain fewer agrochemical residues than conventionally grown alternatives. There is a widespread belief that organic agriculture products are safer and healthier than conventional foods. It is difficult to come to conclusions, but what should be made clear to the consumer is that "organic" does not automatically equal "safe". In the absence of adequate comparative data, additional studies in this area of research are required.     

The interaction of chemicals and viruses and their role in Reye's syndrome

Chemicals have not conclusively been shown to potentiate human viral infections or Reye's Syndrome, but they have been proven to potentiate viral disease in insects, mice, rats, ducks, and tissue in culture. The data suggest that we cannot rule out virus-chemical interactions in the pathogenesis of human disease.Reye's Syndrome (RS), an encephalopathy with fatty changes of the viscera, is a disease of children with a high mortality and unknown etiology. The most frequently suggested causes of RS are viruses or environmental toxins. It has also been suggested that both may cause RS acting either alone or in concert. Varicella or influenza B are the viruses most frequently associated with RS. Aflatoxin, a fungal toxin, has been the toxin most frequently implicated in RS or a disease that is almost identical to it.The hepatic tissue of 9 children with Reye's Syndrome was examined and was found to contain no aflatoxin. The sensitivity of our test was 5.2 μg of aflatoxin B₁/kg wet hepatic tissue. With some reservation, it is felt unlikely that aflatoxicosis is important in the pathogenesis of Reye's Syndrome in the United States.We also examined the serum of 12 patients with RS for pseudocholinesterase activity. Although the mean value of this activity for the RS patients was 12% less than that for the control group, there was no significant differences (P > 0.05) and, therefore, our RS patients did not demonstrate any effects of pseudocholinesterase inhibitors. It is as yet difficult to demonstrate, except coincidentally, environmental toxins in the pathogenesis of Reye's Syndrome.