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Mycotoxins in infant cereal foods from the Canadian retail market.

Three hundred and sixty-three samples of cereal-based infant foods were collected from the Canadian retail marketplace over 3 years. The samples included oat-, barley-, soy-, and rice-based infant cereals, mixed-grain infant cereals, teething biscuits, creamed corn, and soy-based formulas. Samples were analysed for targeted mycotoxins (deoxynivalenol, nivalenol, HT-2 toxin, zearalenone, ochratoxin A, fumonisins B(1) and B(2), and five ergot alkaloids). Soy-based cereals (which usually contain corn) exhibited the highest incidences of deoxynivalenol (100%), zearalenone (46%) and fumonisins (75%). Overall, deoxynivalenol was the most frequently detected mycotoxin--it was detected in 63% of samples analysed. Survey results demonstrated the regular occurrence of multiple mycotoxins in cereal-based infant foods.

Canada↗

Occurrence of mycotoxins and ergosterol in maize harvested over 5 years in Northern Italy.

Maize samples collected from storage bins and feed mills in Northern Italy between 1995 and 1999 were surveyed for the occurrence of aflatoxin B1 (AFB1), zearalenone (ZEA), deoxynivalenol (DON) and fumonisin (FB1); further, ergosterol was analysed as a fungal growth marker. The incidence and mean content of AFB1 were generally low; nevertheless, a remarkable contamination was found in two samples (109 and 158 microg kg(-1)), while five others exceeded 20 microg kg(-1). DON and ZEA mean levels were significantly higher in 1996 (2716 and 453 microg kg(-1)) with respect to the other years, when mean contents ranged from 7 to 30% and from 3 to 17%, respectively, expressed in per cent of 1996 contents. FB1 was present in all samples and was by far the most remarkable mycotoxin in Northern Italian maize, with the exception of samples from 1996. The average level was 3064 microg kg(-1), 69.6% of samples resulted over 1000 microg kg(-1) and 16.9% over 5000 microg kg(-1). Significant correlations were found between ergosterol and the major mycotoxin(s) in each year (FB1 in 1995 and 1997-99; ZEA + DON in 1996). Consequently, ergosterol seems to be a good index of the toxicological quality of maize. Climatic conditions influenced the growth of different fungal species. In 1996, the first 20 days of October were extremely rainy; these weather conditions delayed the harvest until the first week of November and favoured the growth of DON and ZEA producing fungi and the synthesis of mycotoxins. On the contrary, the temperate and dry climate of the other years supported the growth of FB1-producing fungi.

Climate↗

Estimated dietary exposure to principal food mycotoxins from the first French Total Diet Study.

This study reports estimates on dietary exposure from the first French Total Diet Study (FTDS) and compares these estimates with both existing tolerable daily intakes for these toxins and the intakes calculated during previous French studies. To estimate the dietary exposure of the French population to the principal mycotoxins in the French diet (as consumed), 456 composite samples were prepared from 2280 individual samples and analysed for aflatoxins, ochratoxin A, trichothecenes, zearalenone, fumonisins and patulin. Average and high percentile intakes were calculated taking account of different eating patterns for adults, children and vegetarians. The results showed that contaminant levels observed in the foods examined 'as consumed' complied fully with current European legislation. However, particular attention needs to be paid to the exposure of specific population groups, such as children and vegans/macrobiotics, who could be exposed to certain mycotoxins in quantities that exceed the tolerable or weekly daily intake levels. This observation is particularly relevant with respect to ochratoxin A, deoxynivalenol and zearalenone. For these mycotoxins, cereals and cereal products were the main contributors to high exposure.

Adult↗

The role of sampling in mycotoxin contamination: an holistic view.

The need to obtain a representative sample deserves particular consideration since a wrong sampling plan can greatly affect the reliability of the measured levels of mycotoxins. This can even result in legal disputes and barriers to trade. Reported here is an holistic view for an ideal sampling plan, which is based on two consecutive steps: (i) To establish 'why, where and when' sampling has to be performed by assessing the purpose, the appropriate time and the site for collecting the samples; (ii) To establish 'how' to draw samples by assessing practical ad hoc guidelines, considering that, for bulk goods in particular, mycotoxins are not at all homogeneously distributed in a lot. So far, step 1 is not yet covered by specific guidelines while for step 2, European regulations establish the procedures for the sampling of bulk and retail products potentially contaminated by mycotoxins.

Environmental Monitoring↗

Production and characterization of a monoclonal antibody that cross-reacts with the mycotoxins nivalenol and 4-deoxynivalenol.

Nivalenol is a mycotoxin produced by certain fungi that are pathogenic to important cereal crops, in particular maize, wheat, and barley. This toxin, 3alpha,4beta,7alpha,15-tetrahydroxy-12,13-epoxytrichothec-9-en-8-one, is found worldwide and is closely related to 4-deoxynivalenol (DON or vomitoxin), a mycotoxin associated with outbreaks of Fusarium head blight in North America. The literature on the toxicity of nivalenol suggests it is similar, if not more toxic, than DON. Despite the development of rapid immunologically based assays for detecting DON, such assays have not existed for detecting nivalenol without chemical modification of the analyte. This paper describes the development of a monoclonal antibody using a nivalenol-glycine protein conjugate. The monoclonal antibody was most specific for an acetylated form of DON (3-Ac-DON), but it exhibited sensitivity and cross-reactivity that were useful for detecting nivalenol and DON at relevant levels without the need to modify either toxin chemically. In an competitive indirect ELISA format, the concentrations of toxins able to inhibit colour development by 50% (IC50) were 1.7, 15.8, 27.5, 68.9, and 1740 ng ml(-1) for the mycotoxins 3-Ac-DON, DON, nivalenol, 15-Ac-DON, and fusarenon-X, respectively. The antibody was also used to develop a competitive direct ELISA for DON and nivalenol, with IC50's of 16.5 ng ml(-1) (DON) and 33.4 ng ml(-1) (nivalenol). These assays are capable of detecting both DON and nivalenol simultaneously, a property that may be useful in regions where these toxins co-occur or in formats, such as immunoaffinity columns, where co-isolation of both toxins is desirable.

Animals↗

Development and application of analytical methods for the determination of mycotoxins in organic and conventional wheat.

The aim of this study was to develop a multicomponent analytical method for the determination of deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEN), nivalenol (NIV), 3-acetyl-DON (3-acDON), 15-acetyl-DON (15-acDON), zearalenol (ZOL) and citrinin (CIT) in wheat. It also aimed to survey the presence and amounts of DON, OTA and ZEN in Belgian conventionally and organically produced wheat grain and in wholemeal wheat flours. After solvent extraction, an anion-exchange column (SAX) was used to fix the acidic mycotoxins (OTA, CIT), whilst the neutral mycotoxins flowing through the SAX column were further purified by filtration on a MycoSep cartridge. OTA and CIT were then analysed by high-performance liquid chromatography (HPLC) using an isocratic flow and fluorescence detection, while the neutral mycotoxins were separated by a linear gradient and detected by double-mode (ultraviolet light fluorescence) detection. The average DON, ZEN and OTA recovery rates from spiked blank wheat flour were 92, 83 and 73% (RSDR = 12, 10 and 9%), respectively. Moreover, this method offered the respective detection limits of 50, 1.5 and 0.05 microg kg-1 and good agreement with reference methods and inter-laboratory comparison exercises. Organic and conventional wheat samples harvested in 2002 and 2003 in Belgium were analysed for DON, OTA and ZEN, while wholemeal wheat flour samples were taken from Belgian retail shops and analysed for OTA and DON. Conventional wheat tended to be more frequently contaminated with DON and ZEN than organic samples, the difference being more significant for ZEN in samples harvested in 2002. The mean OTA, DON and ZEA concentrations were 0.067, 675 and 75 microg kg-1 in conventional samples against 0.063, 285 and 19 microg kg-1 in organically produced wheat in 2002, respectively. Wheat samples collected in 2003 were less affected by DON and ZEN than the 2002 harvest. Organic wholemeal wheat flours were more frequently contaminated by OTA than conventional samples (p < 0.10). The opposite pattern was shown for DON, organic samples being more frequently contaminated than conventional flours (p < 0.10).

Belgium↗

Natural occurrence of Alternaria mycotoxins in sorghum and ragi from North Bihar, India.

The natural occurrence of major Alternaria mycotoxins i.e. alternariol (AOH), alternariol methyl ether (AME), altenuene (ALT), altertoxin-1 (ATX-1) and tenuazonic acid (TA) has been investigated in sorghum (Sorghum vulgare Pers.) and ragi (Eleusine coracana Gaertn.) collected from North Bihar. Nine out of 20 sorghum samples, and three out of eight ragi samples, were found to be contaminated with one to three Alternaria mycotoxins. The toxin elaborating potential of Alternaria alternata (Fr.) Keissler strains isolated from sorghum and ragi have also been investigated. Out of 16 isolates of A. alternata (Fr.) Keissler, mycotoxins were detected in only 10.

Alternaria↗

Natural occurrence of mycotoxins and mycotoxigenic fungi in cereals in the United Kingdom.

Approximately 50 samples of poorly stored cereals were examined for mycotoxins, fungi and biological activity. While some mycotoxins such as ochratoxin A and citrinin occurred frequently, many other fungal metabolites present could not be identified. Cereal extracts and isolates of some fungi, when grown in culture, were often toxic to brine shrimp larvae but, in some toxic extracts, no known mycotoxins were identified. A preliminary report of this study forms the subject of this paper.

Animals↗

Mycotoxin methodology.

Sensitive, specific, accurate and precise methods of analysis are needed for enforcement of mycotoxin regulations, other monitoring programmes, and research studies. Rapid screening tests are useful for control at all stages of food and feed production. There is a wide choice of both quantitative and qualitative methods for the more well known mycotoxins. Those at present covered by method standardization organizations such as AOAC International are aflatoxins (including M1), Alternaria toxins, citrinin, cyclopiazonic acid, ergot alkaloids, fumonisins, ochratoxins, patulin, trichothecenes, and zearalenone. Methodology for mycotoxins is selectively reviewed in this paper with emphasis on the procedures comprising the analytical method--sampling, extraction of naturally contaminated samples, clean-up, detection and determination, and confirmation. Also covered are automation, method comparison, and method assessment.

Chromatography, High Pressure Liquid↗

Survey of natural occurrence of trichothecene mycotoxins and zearalenone in Korean cereals harvested in 1992 using gas chromatography/mass spectrometry.

For the survey of the natural occurrence of trichothecene mycotoxins, produced by species of fungi imperfecti such as Fusarium and Trichothecium, a sensitive analytical method was developed for the simultaneous detection and quantitation of the major trichothecene mycotoxins, viz. T-2 toxin (T-2), HT-2 toxin (HT-2), nivalenol (NIV), fusarenon-X (F-X), deoxynivalenol (DON), 3-acetyl deoxynivalenol (3-Ac DON), and zearalenone (ZEN), using gas chromatography/mass spectrometry-selected ion monitoring (GC/MS-SIM) mode after trimethyl silyl derivatization. The incidence of NIV and DON in 30 barley samples were 93% and 67%, respectively; the average contents of NIV and DON in positive samples were 390 ng/g (range 40-2038) and 106 ng/g (range 5-361) respectively. In 15 maize samples, the incidences of NIV and DON were 53% and 93% respectively and the average contents were 168 ng/g and 145 ng/g, respectively. These results suggest that NIV and DON were the major contaminating trichothecene mycotoxins in Korean barley and maize samples harvested in 1992.

Edible Grain↗

Determination of the fate of three Fusarium mycotoxins through wet-milling of maize using an improved HPLC analytical technique.

The fate of three Fusarium mycotoxins, nivalenol (NIV), deoxynivalenol (DON) and zearalenone (ZEN), all common contaminants in New Zealand-grown maize, has been measured in fractions of maize after passage through a commercial wet-milling plant. Distribution of the three toxins follows a pattern reasonably expected from their physical solubility characteristics. The highly water-soluble mycotoxins, NIV and DON, were found at high concentrations (up to 8.8 mg/kg) in concentrated steep liquor (CSL) fractions, but at low levels (less than 0.3 mg/kg) in the solid (germ, fibre and gluten) fractions. The converse was true for ZEN, which is relatively insoluble in water. For ZEN, the maximum concentration found in CSL was 0.6 mg/kg compared with 2.2-4.8 mg/kg in germ, fibre and gluten fractions. Accordingly, an animal food byproduct composed mainly of pressed fibre and concentrated steep liquor was usually found to contain concentrations of all three mycotoxins above those existing in the input maize. A single sample of corn oil recovered during the study also had a high concentration (4.6 mg/kg) of ZEN. The analytical clean-up method used converts all trichothecenes present to parent alcohols, therefore results are indicative of total trichothecene content. HPLC analytical conditions suitable for the analysis of NIV and DON in complex process grain products are also described.

Chromatography, High Pressure Liquid↗

Survey of selected mycotoxins in food.

The contamination of a variety of foods with the mycotoxins aflatoxins B1, B2, G1, M1, ochratoxin A, patulin, and byssochlamic acid was investigated. With the developed methods, the identification was made possible by in situ fluorescence spectral analysis with a far reaching exclusion of substances which simulate sycotoxins. Aflatoxin B1 was detected in 2 samples of 105 spontaneously moulded food specimens and only in 1 of a group of 198 food samples that were obviously not moulded. Aflatoxin M1 was identified in 4 of 60 samples of the so-called wintermilk. Investigated dairy products did not contain aflatoxin M1. Ochratoxin A could be detected twice in 49 spontaneously moulded corn samples, aflatoxin, however, was not found. Further investigations of 50 samples of moulded food revealed ochratoxin A in 2 samples of raw coffee. Food samples obviously not moulded did not contain ochratoxin A. Patulin was detected in 19 of 110 samples of fruit and fruit products, especially in commercial apple juice, in rotten parts of apples, and in moulded fruit products. Byssochlamic acid was not detected in fruit juices or in fruit. The toxicological consequences of mycotoxins in food and possibilities to reduce mycotoxins are discussed.

Aflatoxins↗

Influence of organically or conventionally produced wheat on health, performance and mycotoxin residues in tissues and bile of growing pigs.

From 1999-2001 three different varieties of wheat [Contur (susceptible to Fusarium), Batis and Petrus (less susceptible to Fusarium)] were cultivated under organic and conventional conditions in order to determine mycotoxin burden. Soil quality, preceding crop and weather conditions were comparable in the different production systems. The wheat batches were analysed for moulds, and the contents of zearalenone (ZEN) and deoxynivalenol (DON). Feeding trials were carried out with growing pigs (n = 96; average initial live weight 22.2 +/- 1.5 kg [mean +/- SD]) to examine a possible influence on the animal performance and on mycotoxin residues. The data recorded were clinical conditions, performance, biochemical and hematological data. Residues of ZEN, alpha- and beta-zearalenol (ZEL) and of DON were determined in bile, liver and muscle after slaughtering. Conventionally cultivated wheat was more frequently contaminated with Fusarium and contained more frequently ZEN and DON in higher concentrations than the organically produced wheat. Hematological and biochemical parameters of pigs fed with organically cultivated diets were not different from those of conventionally fed pigs. Pigs fed with organically produced wheat showed a slightly higher daily weight gain, but a lower carcass yield than the conventionally fed animals. The highest residues of DON and total-ZEN (ZEN + alpha-ZEL + beta-ZEL) were found in bile. Bile samples of organically fed pigs contained lower concentrations of total-ZEN than those of conventionally fed pigs. Altogether, these data suggest that wheat from an organic farming does not have higher mycotoxin-contamination than wheat from the conventional farming system.

Agriculture↗

Influence of a Fusarium culmorum inoculation of wheat on the progression of mycotoxin accumulation, ingredient concentrations and ruminal in sacco dry matter degradation of wheat residues.

The Fusarium head blight (FHB)-susceptible winter wheat cv. Ritmo was inoculated with spores of Fusarium culmorum at the beginning of full blossom. Samples of whole wheat plants were taken once weekly from anthesis until harvest and subsequently fractionated into straw, glumes and spindles, which were examined for deoxynivalenol (DON) and zearalenone (ZON). Additionally, the content of crude protein (CP) and non-starch polysaccharides (NSP) was scrutinized. Synthesis of the Fusarium toxins DON and ZON generally differed in terms of date of formation and concentration. Final mean DON concentrations of 37.5, 28.1 and 5.0 mg/kg DM were measured in glumes, spindles and straw, respectively, at the time of harvest. At this time, maximal mean ZON concentrations of 587, 396 and 275 microg/kg DM in spindles, glumes and straw, respectively, were determined. Moreover, Fusarium infected wheat residues contained higher CP but lower NSP contents at the last three sampling dates. In addition, collective samples of wheat straw and chaff were taken to investigate the effect of the Fusarium contamination on their in sacco DM degradation in dairy cows. Samples were analysed for mycotoxins and selected quality parameters. The dried and milled collective samples of straw and chaff were weighed into nylon bags and subjected to ruminal incubation for 4, 8, 16, 24, 48, 72, 96 and 120 h in two dairy cows equipped with a permanent rumen cannula. Marked differences in level of mycotoxin contamination as well as in ingredient composition between the variants of straw and chaff were detected. Moreover, after 120 h rumen incubation the in sacco DM degradation of inoculated straw and chaff were lower compared to the accordant controls. The soluble fraction was increased in inoculated samples, whereas a diminishment in the potentially degradable but insoluble fraction was more pronounced. Thereby, a decrease in the potential degradability was obtained for inoculated straw and even if less pronounced for chaff compared to the non inoculated corresponding controls. In conclusion, infection with F. culmorum of wheat involves an increased risk of mycotoxin contamination in straw. Also, a Fusarium infection may have an impact on chemical composition and may result in Fusarium growth-related modifications of host cell wall components.

Animal Feed↗

Effect of feed-borne pollution with some mycotoxin combinations on broiler chicks.

Broiler chicks were divided into five groups and fed starter mash from the first day after hatching. The first group feed (control) was mycotoxin free, whereas the mycotoxins sterigmatocystin (350 ppb) and aflatoxin B1 (100 ppb) were added to the second group diet, patulin (100 ppb) and aflatoxin B1 (100 ppb) to the third group feed, penicillic acid (850 ppb) and aflatoxin B1 (100 ppb) to the fourth group, and aflatoxins B2a (0.9 ppb) +G2 alpha (25 ppb) +M1 (0.9 ppb) +M2 (1 ppb) to the fifth group. This contaminated feeding lasted for four weeks followed by another four weeks as recovery period during which all groups fed finishing mash without mycotoxins. At the end of the experiment, the chickens of groups two, three, four and five were significantly lower in body weight and feed conversion and reflected higher mortality rates than those of the control group.

Animal Feed↗

The initiator tRNA acceptance assay as a short-term test for carcinogens. 4. Results with 20 mycotoxins.

The activity of 20 mycotoxins was tested by the recently developed initiator tRNA acceptance assay for carcinogens. With the exception of citrinin, all compounds carcinogenic for rodents stimulated the charging of tRNA with L-methionine. In three out of four non-carcinogenic mycotoxins the test was negative. Five carcinogenic mycotoxins were negative in mutagenicity tests but positive in the acceptance assay indicating that also non-mutagenic carcinogens may be detected by the latter procedure.

Carcinogenicity Tests↗

Study of the genotoxic potential of 17 mycotoxins with the SOS Chromotest.

Seventeen mycotoxins [aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2), aflatoxicol, sterigmatocystin, patulin, citrinin, penicillic acid, T-2 toxin, diacetoxyscirpenol, zearalenone, zearalenol (alpha and beta isomers 1:1), ochratoxin A, norsolorinic acid, averufin, versicolorin A] were tested using the SOS Chromotest (PQ37 and PQ35). Six of the mycotoxins (AFB1, AFG1, AFB2, aflatoxicol, sterigmatocystin and versicolorin A) were genotoxic on PQ37 strain with and without metabolic activation. The results obtained with metabolic activation are in agreement with positive results obtained in other tests of genotoxicity. Except for AFB2, the presence of a double bond C8-C9 in the dihydrobenzofurane (DHBF) ring explained the activity due to the formation of an epoxide, but the coumarin cyclopentenone ring also plays a role in the qualitative differences of genotoxic activity. The wild-type uvrB gene in PQ35 decreases the genotoxic response with and without metabolic activation. Without metabolic activation, only mycotoxins possessing the DHBF ring group and double linkage C8-C9 exhibit a genotoxic effect.

Animals↗

Effects of feeding blends of grains naturally contaminated with Fusarium mycotoxins on production and metabolism in broilers.

Three hundred sixty, 1-d-old male broiler chicks were fed diets containing grains naturally contaminated with Fusarium mycotoxins for 56 d. The four diets included control (0.14 mg/kg deoxynivalenol, 18 mg/ kg fusaric acid, < 0.1 mg/kg zearalenone), low level of contaminated grains (4.7 mg/kg deoxynivalenol, 20.6 mg/kg fusaric acid, 0.2 mg/kg zearalenone), and high level of contaminated grains without (8.2 mg/kg deoxynivalenol, 20.3 mg/kg fusaric acid, 0.56 mg/kg zearalenone) and with (9.7 mg/kg deoxynivalenol, 21.6 mg/kg fusaric acid, 0.8 mg/kg zearalenone) 0.2% esterified-glucomannan polymer derived from Saccharomyces cerevisiae1026 (E-GM). Body weight gain and feed consumption responded in a significant quadratic fashion to the inclusion of contaminated grains during the finisher period. Efficiency of feed utilization, however, was not affected by diets. The feeding of contaminated grains in the finisher period also caused significant linear increases in blood erythrocyte count and serum uric acid concentration and a significant linear decline in the serum lipase activity. Dietary inclusion of contaminated grains resulted in a significant quadratic effect on serum albumin and y-glutamyltransferase activity. Blood hemoglobin and biliary IgA concentrations, however, responded in significant linear and quadratic fashions. Supplementation of E-GM counteracted most of the blood parameter alterations caused by the Fusarium mycotoxin-contaminated grains and reduced breast muscle redness. It was concluded that broiler chickens may be susceptible to Fusarium mycotoxicoses when naturally contaminated grains are fed containing a combination of mycotoxins.

Animal Feed↗