Aflatoxin M1 contamination in commercial pasteurized milk from local markets in Fariman, Iran

Contamination of milk and dairy products with aflatoxin M₁ (AFM₁) presents a risk for human health. The aim of this study was to investigate the presence of AFM₁ in pasteurized milk samples in Fariman, located in the province of Khorasan Razavi, Iran, by enzyme-linked immunosorbent assay (ELISA). Forty-five samples of pasteurized milk from different supermarkets were collected during 3 months in summer (July to September, 2012). AFM₁ contamination was detected in all of milk samples. The mean concentration of aflatoxin M₁ was 27.2 ng/l. The range of AFM₁ content was 8.8–64 ng/l. Thirteen (28.8 %) of the samples had AFM₁ levels exceeding the maximum levels (50 ng/l) accepted by the European Union. Due to the fact that milk is used by all the age groups including infants and children in Fariman city, it is necessary to minimize the health risk from AFM1 contamination in milk. For this reason, the level of its precursor, aflatoxin B₁ (AFB₁), in dairy feeds must be reduced, requiring constant aflatoxin monitoring of relevant agricultural commodities.     

In vivo covalent binding of aflatoxin B1 and aflatoxin M1 to liver DNA of rat,mouse and pig

[¹⁴C]Aflatoxin B₁ (AFB₁) was isolated from cultures of Aspergillus parasiticus grown on [1-¹⁴C]sodium acetate. Covalent binding of AFB₁ to liver DNA of rat and mouse was determined 6–8 h after oral administration. The effectiveness of covalent binding, expressed as DNA binding per dose in the units of a ‘Covalent Binding Index’ (CBI), (μmol aflatoxin/mol DNA nucleotides)/(mmol aflatoxin/kg animal), was found to be 10 400 for rats and 240 for mice. These CBI partly explain the different susceptibility of the two species for the incidence of hepatic tumors.The corresponding values for pig liver DNA, 24 and 48 h after oral administration, were found to be as high as 19 100 and 13 300. DNA-binding has not so far been reported for this species although it could represent an appropriate animal model for studies where a human-like gastrointestinal tract physiology is desirable.Aflatoxin M₁ (AFM₁) is a metabolite found in the milk of cows that have been fed AFB₁-contaminated diet. [¹⁴C]AFM₁ was also found to be produced by cultures of A. parasiticus giving a yield of about 0.3% of the total aflatoxins. A test for covalent binding to rat liver DNA revealed a CBI of 2100 showing that AFM₁ must also be regarded as a strong hepatocarcinogen. It is concluded that AFB₁ contaminations should be avoided in dairy feed.     

Occurrence of aflatoxin M<Subscript>1</Subscript> in commercial pasteurized milk samples in Sari,Mazandaran province,Iran

The frequency and levels of aflatoxin M₁ (AFM₁) in pasteurized milk samples in Sari, located in Mazandaran province, Iran, were determined by enzyme immunoassay. Seventy-six samples of pasteurized milk from different retail stores were randomly collected over four seasons during the year 2015. AFM₁ contamination was detected in all milk samples. The mean concentration of aflatoxin M₁ was 65.8 ng/l, with a range of 11.7–106.6 ng/l. The highest AFM₁ level was detected in milk samples collected during spring. Forty-six (60.53 %) samples had AFM₁ levels that exceeded the maximum acceptable levels (50 ng/l) recommended by the European Union (EU). Comparison of these results with previously published data for AFM₁ in milk in Iran shows that the percentage of samples exceeding the EU maximum level is consistently high over the years, indicating a general problem related to AFB₁ contamination in dairy feedingstuff.     

Reduction in the urinary aflatoxin M1 biomarker as an early indicator of the efficacy of dietary interventions to reduce exposure to aflatoxins

Abstract

Aflatoxin B₁ is a persistent public health issue in Ghana. Assessment of AFB₁ intervention efficacy is currently dependent on long-term biomarkers. This study was designed to determine whether daily AFM₁ biomarker levels could be utilized as an early detection method for intervention efficacy. Participants were treated with a refined calcium montmorillonite clay (UPSN) or a placebo (calcium carbonate) in a crossover study. Urine samples were assessed for AFM₁ levels daily. UPSN treatment reduced AFM₁ biomarkers by 55% compared to the placebo. This is the first study to show that daily urinary AFM₁ levels can be used as a biomarker of internal aflatoxin B₁ exposure in short-term intervention trials to determine efficacy.     

Mycotoxins in South African foods: a case study on aflatoxin M<Subscript>1</Subscript> in milk

The contamination of cow’s milk at the farm level with aflatoxin M₁ was investigated in South Africa. Samples of feeds, forage, maize and milk were taken at nine dairy farms, and at the same time samples of the processed milk (retail milk) were collected from the respective dairies to which the farms delivered their milk. The feeds were analysed for aflatoxin B₁ and the milk samples for aflatoxin M₁ using high performance liquid chromatography (HPLC) and fluorescence detection. All milk samples from the dairy farms were positive for aflatoxin M₁, ranging from 0.02 μg/l to 1.5 μg/l. Retail milk was also frequently contaminated with AFM₁, at levels of 0.01-3.1 μg/l. High AFB₁ levels in feed materials on the farms supplying the raw milk indicate that various sources account for this contamination frequency in milk.     

Aflatoxin M<Subscript>1</Subscript> levels in different marketed milk products in Nairobi,Kenya

Milk is an important source of energy and nutrients, especially for children, and in Kenya, milk consumption is higher than other countries in the region. One major concern with milk is the risks of chemical contaminants, and reports of high levels of aflatoxin M₁ (AFM₁) in milk in Kenya has been causing public health concerns. This study collected marketed milk products every month during 1 year, just as a consumer would purchase them from retailers and traders in a low-income area, and a major supermarket in a middle/high-income area. In total, 291 sampled milk products (raw, pasteurised, UHT milk, yoghurt and lala) were collected and analysed for AFM₁ using a commercial ELISA kit. More than 50% of the samples exceeded 50 ng/kg (the level allowed in the EU), but only three samples exceeded 500 ng/kg (the level allowed in the USA). Geometric mean AFM₁ level was 61.9 ng/kg in the 135 samples from the low-income area while it was 36.1 ng/kg in the 156 from the higher income area (p?<?0.001). The levels varied significantly depending on the time of year, with lowest levels of milk in January. There were also differences between manufacturers and products, with UHT milk having lower levels. There was no difference depending on the price for all dairy products, but when only including milk, higher price was associated with lower levels of AFM₁. In conclusion, this study shows that milk purchased by a consumer is likely to contain AFM₁ above 50 ng/kg, and that further research is needed to find ways to mitigate AFM₁ contamination through working with farmers and milk processors both in the formal and informal sectors.     

A six year survey (1999–2004) of the ocurrence of aflatoxin M1 in daily products produced in Portugal

Aflatoxin M₁, a mutagenic and carcinogenic metabolite of aflatoxin B₁, occurs in milk from animals fed on food contaminated with some species ofAspergillus. Aiming to investigate the occurrence of AFM in dairy products produced in Portugal. 598 samples of raw milk were analysed during six years (1999–2004). 25 samples of powder milk and 42 traditional fresh cheeses were also analysed. The toxin was extracted using an immunoaffinity column method and quantified by HPLC. AFM₁ was detected in 394 (65.8%) of the raw milk samples. Along the analysed period AFM: was detected at a low level (0.005–0.05μg/l) in 54.8% of the samples and at a level ranging from 0.041–0.05 in 2.8% of the samples. From 2001 to 2004, 49 samples (8.2%) were contaminated with levels above the maximum permitted level (>0.051 to 0.08). None of the samples of powder milk and traditional fresh cheese revealed to be contaminated with AFM₁.     

Vaccination of Heifers with Anaflatoxin Improves the Reduction of Aflatoxin B1 Carry Over in Milk of Lactating Dairy Cows

It was previously reported that injection of anaflatoxin B₁ (AnAFB₁) conjugated to keyhole limpet hemocyanin (KLH), together with Freund''s adjuvant, was effective in inducing in cows a long lasting titer of anti-aflatoxin B₁ (AFB₁) antibodies (Abs), cross-reacting with other aflatoxins, which were able to hinder, proportionally to their titer, the secretion of aflatoxin M₁ (AFM₁) into the milk of cows continuously fed with AFB₁. According to anti-AFB₁ Ab titer, 50% of the vaccinated cows were recognized as high responder animals. In an attempt to prepare a more effective formulation for vaccination of cows, it was compared the immunogenicity, in Holstein Friesian heifers, of AnAFB₁ covalently conjugated to KLH or to recombinant diphtheria toxin (CRM₁₉₇) molecules, and injected together with various adjuvants. This study demonstrated that injection of AnAFB₁ conjugated to KLH and mixed with complete (priming) and incomplete Freund''s adjuvant (boosters), as in the previous schedule of immunization, was the most effective regimen for inducing Ab responses against AFB₁, although pre-calving administration could increase the effectiveness of vaccination, resulting in 100% high responder animals. After one booster dose at the beginning of the milk production cycle, anti-AFB₁ Ab titers were comparable to those recorded at the end of the immunization schedule, and proved to be effective in reducing significantly AFB₁ carry over, as AFM₁, from feed to milk. Pre-calving vaccination of dairy heifers with conjugated AnAFB₁, adjuvated with complete and incomplete Freund''s adjuvant, may represent the most effective tool for preventing the public health hazard constituted by milk and cheese contaminated with aflatoxins.     

Isolation and molecular characterization of mycotoxigenic fungi in agarwood

Agarwood (Oudh), is often used by people in the Kingdom of Saudi Arabia. The Oudh has been mentioned in the Hadith and is traditionally used for its aroma (perfuming smell) and potential medicinal applications. The aim of the study was to isolate mycotoxigenic fungi that grow on agarwood and the factors and storage conditions that enhance their growth potential. In addition to the detection of associated mycotoxins like: Aflatoxin B1 (AFB₁) and ochratoxin A (OTA) from agarwood. Agarwood samples were collected from local markets of Jeddah governorate, Kingdom of Saudi Arabia. Standard dilution plate method was used for the isolation of fungi. Isolated fungi were identified based on morphological characteristics and confirmed using molecular biology techniques. AFB₁ and OTA were detected by High Performance Liquid Chromatography (HLPC). The results indicated that the most commonly isolated fungal genera were in the following descending order: Aspergillus, Penicillium, Fusarium and Rhizopus. Among Aspergillus genera, A. flavus and A. ochraceus were detected based on their morphology and confirmed by PCR using specific primers. It was also noted that AFB₁ was released by 15.3 and 55.0% of A. flavus and A. parasiticus isolates respectively with levels reaching up to 14.60 µg/L. The moisture content in the samples ranged from 3% to 10% affected fungal growth. AFB₁ was detected in 22 out of 50 of the samples. The maximum level of AFB₁ (50.7 µg/kg) was detected in samples with higher moisture content (12%) stored at a temperature of 32 °C. Aspergillus fungi were found to be the most predominant fungal genera found on agarwood. Moisture content (9–10%) and storage temperature (32 °C) stimulated fungal growth and their ability to produce mycotoxins. For this reason, storage conditions at the marketing place should be adequate in order not to provide a conducive environment for fungal growth which is associated with the mycotoxin production. In order to prevent fungal growth and mycotoxin production, it would be recommended to store agarwood at temperatures not exceeding 25 °C and moisture content of up to a maximum of 5–6%.     

Invitro metabolism of aflatoxin B1 by rat liver nuclei

Rat liver nuclei were used to study the metabolism of aflatoxin B₁ (AFB₁). Nuclei were found to metabolize AFB₁ to aflatoxins M₁(AFM₁), Q₁(AFQ₁), and P₁(AFP₁); the formation of AFP₁ was relatively negligible. AFM₁ was preferentially formed when rats were pretreated with 3-methylcholanthrene (MC), whereas phenobarbital (PB) pretreatment enhanced the formation of both AFM₁ and AFQ₁. PB pretreatment also resulted in increased binding of AFB₁ metabolites to DNA and to other macromolecules in the nucleus. Following PB and MC pretreatment, induction specificities of the nuclear metabolism of AFB₁ were similar to those of the microsomal metabolism.     

Limited survey on aflatoxin contamination in rice

Aflatoxins (AFS) are toxic and carcinogenic fungal metabolites. Aflatoxin B1 is the most toxic and has been classified as a Group I carcinogen by the International Agency for Research on Cancer (IARC). Samples of imported rice were analyzed for their AFS content. Finley ground rice subsamples were extracted with water/methanol (100:150 v/v) followed by purification with Immunoaffinity columns (IAC). AFS purified from extracts were determined with RP-HPLC-FLD using post column electrochemical derivatization with a Kobra Cell. Concentrations of aflatoxin B1 and total AFS in test rice samples were ≤0.123 and ≤2.58 µg/kg, respectively. Tween 80 improved recoveries (86 and 106%) of aflatoxin B1 and aflatoxin G1 from brown rice. Recoveries of Aflatoxin B2 and aflatoxin G2 were substantially reduced (non-detected to 27%) by Tween 80 used in IAC cleanup of brown rice extracts. Visible dense growth of Aspergillus parasiticus (food isolate) occurred at 25 °C but higher aflatoxin B1amounts (23.9–39.3 µg/kg) accumulated when the mold grew at 37 °C in rice seeds stored for three weeks. It could be concluded that levels of aflatoxin B1 and total AFS in rice samples were within the permissible amounts of the EU and other international legislations.     

Aflatoxin M1 in breast milk of nursing Sudanese mothers

The presence of aflatoxin M₁ (AFM₁) in the breast milk of nursing Sudanese mothers was investigated using AOAC official method 980.21 as the extraction method and HPLC with fluorescence detector for separation and detection. Following informed consent, 94 breast milk samples of mothers were collected, and 51 samples were found to be positive for AFM₁, with an average concentration of 0.401?±?0.525?ng?g^?1 and a maximum level of 2.561?ng?g^?1. The volunteers completed a questionnaire concerning their dietary preferences. The data collected suggest that peanut butter, vegetable oils and rice are the main sources responsible for the AFM₁ burden in breast milk. The toxin levels are alarmingly high, and indicate that Sudanese infants are exposed to high levels of AFM₁. A wide range of harmful effects, and consequently health problems, can be expected due AFM₁ toxicity.     

Aflatoxin M<Subscript>1</Subscript> in human breast milk in southeastern Turkey

This study was performed to determine aflatoxin M₁ (AFM₁) in human breast milk samples collected in ?anl?urfa, located in Southeastern region of Turkey, and to investigate a possible correlation between AFM₁ occurrence (frequency and levels) and sampling seasons. Human breast milk samples collected in December 2014 and in June 2015 from a total of 74 nursing women, both outpatient and inpatient volunteers in hospitals located in ?anl?urfa, Turkey, were analyzed using competitive enzyme-linked immunosorbent assay (ELISA) for the presence of AFM₁. AFM₁ was detected in 66 (89.2%) out of 74 samples at an average concentration of 19.0 ± 13.0 ng/l (min.-max., 9.6–80 ng/l). There was a statistically significant difference between December and June concerning AFM₁ levels (p < 0.05). Further detailed studies will be needed to determine the main sources of aflatoxins in food, to establish protection strategies against maternal and infant exposure to these mycotoxins.     

Survey of aflatoxin M1 in cows’ milk from free-grazing cows in Abeokuta,Nigeria

Aflatoxin M₁ (AFM₁) in milk from 100 different herds of free-grazing cows in Abeokuta, Nigeria, was analysed by immunoaffinity column cleanup and HPLC with fluorescence detection. AFM₁ was found in 75 % of the samples, the toxin levels in positive samples ranged from 9.0 to 456.0 ng/l. The mean AFM₁ level in positive samples was 108.15 ng/l, exceeding, for example, the European Union maximum level by a factor of two. These results indicated that there is an urgent need to more closely control the milk of free-grazing cows for AFM₁ in order to protect the health of humans consuming milk and milk products.     

Predictive formulae for goat cheese yield based on milk composition

Prediction of the yield and quality of different types of cheeses that could be produced from a given type and/or amount of goat milk is of great economic benefit to goat milk producers and goat cheese manufacturers. Bulk tank goat milk was used for manufacturing hard, semi-hard and soft cheeses (N = 25, 25 and 24, respectively) to develop predictive formulae of cheese yield based on milk composition. Fat, total solids, total protein and casein contents in milk and moisture-adjusted cheese yield were determined to establish relationships between milk composition and cheese yield. Soft, semi-hard and hard cheeses in this study had moisture contents of 66, 46 and 38%, respectively, which could be used as reference standards. In soft cheese, individual components of goat milk or a combination of two or three components predicted cheese yield with a reasonably high correlation coefficient (R² = 0.73–0.81). However, correlation coefficients of predictions were lower for both semi-hard and hard cheeses. Overall, total solids of goat milk was the strongest indicator of yield in all three types of cheeses, followed by fat and total protein, while casein was not a good predictor for both semi-hard and hard cheeses. When compared with moisture-adjusted cheese yield, there was no difference (P > 0.05) in predicting yield of semi-hard and hard goat milk cheeses between the developed yield formulae in this study and a standard formula (the Van Slyke formula) commonly used for cow cheese. Future research will include further validation of the yield predictive formulae for hard and semi-hard cheeses of goat milk using larger data sets over several lactations, because of variation in relationships between milk components due to breed, stage of lactation, season, feeding regime, somatic cell count and differences in casein variants.     

Presence of adropin,nesfatin-1, apelin-12, ghrelins and salusins peptides in the milk,cheese whey and plasma of dairy cows

Biological fluids (milk and serum/plasma) and cheese whey milk-derived fluid contain numerous molecules, especially amino acids and proteins. Therefore, the purpose of this study was to find out whether cheese whey (n:6), cow milk (n:6) and its blood (n = 6) have adropin, nesfatin-1, apelin-12, ghrelins and salusin peptides. Adropin, nesfatin-1, apelin-12 concentrations were measured by ELISA, whereas ghrelin and salusin concentrations were measured by EIA methods. It was found that adropin, nesfatin-1, apelin-12, des-acylated ghrelin and salusins in cheese whey were higher than in the corresponding milk peptides and plasma of dairy cows, with the exception of salusin alpha and acylated ghrelin in milk being the same than that of the corresponding cheese whey concentration and plasma of dairy cows. A correlation was also found between milk peptides and cheese whey, as also with plasma of dairy cows. The data suggest that peptides in cow milk might be an important and nutritious food for (neonatal) calves and human diet due to their biological and physiological properties.     

Dietas ricas en grasa y composición corporal a lo largo de dos generaciones. Estudio experimental

Introduction and objectiveDespite recent findings reported on the nutritional factors that induce epigenetic changes, little information is available at early ages. This study analyzed in an experimental model, over two generations, potential changes in body composition and potential expression of epigenetic changes as the result of the intake of isoenergetic diets with different fat levels.Materials and methodsAt weaning, Wistar female rats were divided into two groups that were fed either a control diet (fat = 7% w/w) or a high-fat diet (15% w/w). Rats were mated at 70 days (M₁) and their pups (P₁) were the first generation; P₁ rats were mated at 70 days (M₂) and their pups (P₂) represented the second generation. At weaning, mothers and pups (M₁, M₂ and P₁, P₂) were measured body weight (W) and composition (% body fat, %BF), and total skeleton bone mineral content (BMC), expressed as %BMC, using chemical and DXA methods respectively.ResultsAt weaning, high-fat diet groups M₂ and P₂ showed significant increases in W and %BF (p < 0.05); increased %BF values were already found in the M₁ and P₁ groups (p < 0.001). By contrast, %BMC significantly decreased in M₂ and P₂ rats (p < 0.001).ConclusionThis study demonstrates the need to review certain eating habits to avoid perpetuation of unhealthy patterns generation after generation.     

TEP/TDM 18F-choline dans les récidives biologiques des cancers de la prostate traites par radiothérapie externe ou curiethérapie : impact du PSA et de sa cinétique

AimTo determine the impact of PSA and its kinetics on ¹⁸F-Choline PET/CT (FCH PET) ability to detect site of relapse in prostate cancer initially treated with external beam radiotherapy (EBRT) or brachytherapy (IBT).MethodsWe retrospectively enrolled PET FCH performed for suspicion of biochemical relapse after EBRT/IBT from January 2010 to January 2017 at Institut Curie. PSA_trigger, ΔPSA_nadir (PSA_trigger-PSA_nadir), PSA doubling time (PSA_dt) and velocity (PSA_vel) were compared between positive and negative results. Logistic regression analysis was used to determine the relationship between these parameters and PET ability to detect True Positives (TP).ResultsIn all, 271 FCH PET met the inclusion criteria: 169 after treatment with EBRT and 102 after IBT. Positivity rate was 67.9%, and 63.4% of TP were local relapses. Overall sensitivity and specificity were 81.2% and 71.0%. PSA_trigger was 3.32 ng/mL (interquartile space: IQS 2.28–5.77) when PET was negative and 5.15 ng/mL (IQS 3.16–10.17) when positive, ΔPSAnadir was respectively 2.76 ng/mL (IQS 1.84–4.69) and 4.57 ng/mL (IQS 2.48–8.85), PSA_dt 10.78 months (IQS 5.46–20.07) and 7.23 months (EI 2.58–14.14), and PSA_vel 2.16 ng/mL/year (EI 1.02–4.80) et 4.92 ng/mL/year (1.89–16.02) (P < 0.001). Positivity rate increased with PSA_trigger and ΔPSA_nadir. We found PSA_dt ≤ 9 months (P = 0.029; OR = 2.97, IC_95% [1.12–7.88]) and ΔPSA_nadir ≥ 3 ng/mL (P = 0.03; OR = 2.56, IC_95% [1.37–4.77]) to be independent predictive factors of PET sensitivity.ConclusionDetection of relapse after EBRT or IBT with PET FCH is influenced by PSA and its kinetics. In our study, PSA_dt and ΔPSA_nadir were independant predictors of PET performance, but initial treatment and tumor characteristics were not.     

Study of aflatoxin B1 production by Aspergillus parasiticus in bee pollen of Greek origin

Aflatoxin B₁ (AFB₁) is a carcinogenic metabolite produced by certain Aspergillus species such as A. parasiticus and A. flavus. The beneficial properties of bee pollen have transformed this commodity into an increasingly frequent component of the human diet. As bee pollen is a substrate on which aflatoxigenic fungi can grow, AFB₁ production is likely. In the present study, we describe a method for aflatoxin B₁ determination in bee pollen utilising high pressure liquid chromatography (HPLC) with a fluorescence detector (FD). The recovery factor of the method was found to be 111% (RSD% 1.61), while the detection limit (LOD) was 0.08 ng AFB₁/g. An additional aim of this study was to investigate the growth of A. parasiticus and AFB₁ production in bee pollen. Results indicated that no mycelial growth was observed and no AFB₁ was detected in bee pollen samples containing natural microbiota throughout the entire observation period (20 days). In contrast, AFB₁ production in treated bee pollen samples (15 g pollen/flask) inoculated with A. parasiticus was significantly higher (p?≤?0.05) compared to control samples (treated but not inoculated) throughout the entire incubation period, while no mycelial growth was apparent. Maximum production was observed on the 12th day (79.29 ng AFB₁/flask and 32.44 ng AFB₁/flask for inoculated and non-inoculated bee pollen, respectively). As a result, AFB₁ production in bee pollen is likely even following a minor contamination, which could occur randomly.     

Preparation, purification and characteristics of an aflatoxin degradation enzyme from Myxococcus fulvus ANSM068

Aims: To prepare, purify and characterize an extracellular enzyme from Myxococcus fulvus ANSM068, designated as myxobacteria aflatoxin degradation enzyme (MADE), which possesses degradation activity against aflatoxin B₁ (AFB₁), G₁ (AFG₁) and M₁ (AFM₁) in solution. Methods and Results: The culture supernatant of strain M. fulvus demonstrated high degradation ability against AFB₁ (71·89%), AFG₁ (68·13%) and AFM₁ (63·82%) after 48 h of incubation. An enzyme was purified from the supernatant of M. fulvus using ethanol precipitation and chromatography on DEAE‐Sepharose and Superdex 75. An overall 166‐fold purification of the enzyme with a recovery of 57% and a final specific activity of 569·44 × 10³ U mg^?1 was obtained using the present purification protocol. The apparent molecular mass of MADE was estimated to be 32 kDa by SDS‐PAGE. AFG₁ and AFM₁ were significantly degraded, by 96·96 and 95·80%, respectively, when treated with pure MADE (100 U ml^?1) produced by strain ANSM068. MADE exhibited the largest amount of activity at 35°C and pH 6·0, with Mg²⁺ ions greatly promoting and Zn²⁺ strongly inhibiting MADE activity. Conclusions: An aflatoxin degradation enzyme from bacterial isolates can effectively remove aflatoxin B₁, G₁ and M₁ in solution. Significance and Impact of the Study: The high activity and wide temperature and pH range of MADE for the degradation of aflatoxin have promising applications in control of mycotoxins during food and feed processing.