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Fusarium species infect maize crops leading to the production of fumonisin by their toxigenic members. Elimination of microbes is critical in mitigating further postharvest spoilage and toxin accumulation. The current study investigates the efficacy of a previously described multispectral sorting technique to analyze the reduction of fumonisin and toxigenic Fusarium species found contaminating maize kernels in Kenya. Maize samples (n = 99) were collected from six mycotoxin hotspot counties in Kenya (Embu, Meru, Tharaka Nithi, Machakos, Makueni, and Kitui County) and analyzed for aflatoxin and fumonisin using commercial ELISA kits. Aflatoxin levels in majority (91%) of the samples were below the 10 ng/g threshold set by the Kenya Bureau of Standards and therefore not studied further. The 23/99 samples that had >2,000 ng/g of fumonisin were selected for sorting. The sorter was calibrated using kernels sourced from Ghana to reject visibly high-risk kernels for fumonisin contamination using reflectance at nine distinct wavelengths (470-1,550 nm). Accepted and rejected streams were tested for fumonisin using ELISA, and the presence of toxigenic Fusarium using qPCR. After sorting, there was a significant (p < 0.001) reduction of fumonisin, by an average of 1.8 log ng/g (98%) and ranging between 0.14 and 2.7 log ng/g reduction (28-99.8%) with a median mass rejection rate of 1.9% (ranged 0% to 48%). The fumonisin rejection rate ranged between 0 and 99.8% with a median of 77%. There was also a significant reduction (p = 0.005) in the proportion of DNA represented by toxigenic Fusarium, from a mean of 30-1.4%. This study demonstrates the use of multispectral sorting as a potential postharvest intervention tool for the reduction of Fusarium species and preformed fumonisin. The spectral sorting approach of this study suggests that classification algorithms based on high-risk visual features associated with mycotoxin can be applied across different sources of maize to reduce fumonisin., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Asha B. Mohamed reports financial support was provided by International Atomic Energy Agency. Editorial Board member of the Journal of Food Protection – MJS., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Abstract: A total of 133 samples of whole wheat and barley grains and wheat and barley flour collected from retail markets in the main cities of Punjab, Pakistan, were analyzed for the mycotoxin fumonisin B1 (FB1) using reverse phase high-performance liquid chromatography with fluorescence detection. Of these samples, 120 (90%) were positive for FB1, and 75 (63%) of the 120 positive samples had FB1 concentrations higher than the European Union maximum (200 μg/kg). The limit of detection was 4 μg/kg. The highest mean (±SD) concentration of FB1 was found in whole wheat samples, 980.5 ± 211.4 μg/kg. The calculated dietary intakes of FB1 from wheat and barley flours were 4,456 and 503.7 ng/g of body weight per day, respectively., (Copyright ©, International Association for Food Protection.)

Maize ( Zea mays) is a staple in many developing countries but is known to be prone to pest (insects, birds, and rodents) and fungal infestation. In Guatemala, mycotoxin contamination of cultivated products may occur owing to such factors as environmental conditions and the use of traditional agriculture operations. To assess the current maize conditions in Guatemala, a small-scale study was performed. Mold and insect counts and mycotoxin (aflatoxin and fumonisin) concentrations were determined on 25 farms in two townships (Chiantla and Todos Santos) of the Huehuetenango Department. Total fungal counts were 3.6 to 6.83 log CFU/g with no significant differences ( P > 0.05) across farms at different altitudes. Farms where maize was not produced but was purchased were at higher risk of fumonisin contamination, whereas local producers were mostly affected by aflatoxins. Aflatoxin was present in maize from 100% of farms at 1.0 to 85.3 ppb, and fumonisin was detected on 52% of farms at 0.4 to 31.0 ppm. Average mycotoxin consumption amounts were above the recommended maximum intake for aflatoxin in both produced and purchased maize and above the provisional maximum tolerable daily intake for fumonisin in purchased maize. Estimated daily intake was 0.01 to 0.85 μg/kg of body weight per day for aflatoxin and 2.9 to 310.0 μg/kg of body weight per day for fumonisin. An entomological analysis revealed overall 32% prevalence of Ephestia kuehniella (flour moth), 16% prevalence of Sitophilus zeamais (maize weevil), and 8% prevalence of Tribolium sp. (flour beetle) on the analyzed farms. This study highlighted poor agricultural practices used in the highlands of Guatemala. Current practices should be revised for the production of maize that is safe for consumption by the population in this region.

The genus Aspergillus section Nigri, or the black aspergilli, represents genetically closely related species that produce the mycotoxins, ochratoxins and the fumonisins. Fumonisin B1 (FB1) is of an added concern because it is also a virulence factor for maize. Our preliminary data indicated that black aspergilli could develop asymptomatic infections with maize and peanuts plants. Symptomless infections are potential problems, because under favorable conditions, there is a potential for accumulation of ochratoxins and the fumonisins in contaminated postharvest crops. In the present report, the ability of black aspergilli from peanuts and maize to produce ochratoxin A and FB1 on maize kernels was assessed. One hundred fifty strains from peanuts and maize were isolated from several southeastern and midwestern states. Aspergillus nigri (A. nigri var. nigri) was the dominant species (87%), while Aspergillus foetidus, Aspergillus japonicus, Aspergillus tubingensis, and Aspergillus carbonarius were infrequently isolated. None of the wild isolates produced detectable amounts of ochratoxins. However, we do report the occurrence of the fumonisins B1, B2, and B3. Of 54 field isolates, 30% (n = 16) produced FB1, 61% (n = 33) produced FB2, and 44% (n = 24) produced FB3. The amounts of fumonisins produced during the test period of 30 days suggest that these strains might be weak to moderate producers of fumonisin on maize. To our knowledge, this is a first report of FB1 and FB3 production by isolates of black aspergilli from an American cereal and legume.

Powdery mildew (PM), caused by the fungus Erysiphe necator, is one of the most widespread fungal disease of grape and may cause extensive openings on the berry surface during the infection. We evaluated the effect of damage caused by PM in grape berries on the growth of and mycotoxin production by Aspergillus and on the oxidative stress in infected berries. Berries of Vitis vinifera L. cv. Negroamaro with sound skin (SS) and those naturally infected by PM were surface sterilized and inoculated with either fumonisin B2(FB2)-producing strains of Aspergillus niger or ochratoxin A (OTA)-producing strains of Aspergillus carbonarius and incubated at 20 and 30°C. The PM berries were significantly more susceptible to both Aspergillus colonization (5 to 15 times more susceptible) and OTA and FB2 contamination (2 to 9 times more susceptible) than were SS berries. The highest toxin concentration was detected in inoculated PM berries both for OTA (9 ng/g) at 20°C and for FB2 (687 ng/g) at 30°C. In inoculated SS and PM berries, although malondialdehyde and hydrogen peroxide concentrations did not increase, the two black Aspergillus species caused a significant decrease in ascorbate content, thus inducing a pro-oxidant effect. These results indicate that grape berries affected by PM are more susceptible to black Aspergillus growth and to production and/or accumulation of FB2 and OTA. Thus, preventive control of E. necator on grape berries could reduce the mycotoxicological risk from black Aspergillus infection.

In a previous study, inedible almond pick-out samples were assayed for aflatoxin and aflatoxigenic Aspergillus species. These samples contained high populations of black-spored Aspergillus section Nigri species. To investigate whether these species may contribute to the total potential mycotoxin content of almonds, Aspergillus section Nigri strains were isolated from these samples and assayed for ochratoxin A (OTA) and fumonisin B2 (FB2). The majority of isolates (117 strains, 68%) were identified as Aspergillus tubingensis, which do not produce either mycotoxin. Of the 47 Aspergillus niger and Aspergillus awamori isolates, 34 strains (72%) produced FB2 on CY20S agar, and representative strains produced lower but measurable amounts of FB2 on almond meal agar. No OTA-producing strains of Aspergillus section Nigri were detected. Almond pick-out samples contained no measurable FB2, suggesting that properly dried and stored almonds are not conducive for FB2 production by resident A. niger and A. awamori populations. However, 3 of 21 samples contained low levels (<1.5 ng/g) of OTA, indicating that sporadic OTA contamination may occur but may be caused by OTA-producing strains of other Aspergillus species.

A total of 255 corn samples collected in 2010 from three main corn production provinces of China (Liaoning, Shandong, and Henan) were analyzed for contamination with fumonisins (FB(1) and FB(2)). The incidence of contamination was significantly higher in samples from Liaoning than in samples from the other two provinces. Approximately 80.0% of the samples from Liaoning were contaminated with fumonisins, with a mean total fumonisin concentration of 3,990 ng/g. In contrast, the mean total fumonisin concentrations were 845 and 665 ng/g in samples from Shandong and Henan, respectively. The probable daily intake of fumonisins (0.3 μg/kg of body weight) is within the provisional maximum tolerable daily intake of 2.0 μg/kg of body weight set by the Joint Food and Agriculture Organization and World Health Organization Expert Committee on Food Additives.

Fungi belonging to Aspergillus section Nigri occur frequently and in high populations on grapes. Species within this section include Aspergillus niger, A. tubingensis, and A. carbonarius, and they are potential sources for mycotoxins including ochratoxin A and fumonisin B(2) (FB(2)) in grapes and grape products. Aspergillus section Nigri strains were isolated from California raisins to examine the frequency and extent of FB(2) production. Of 392 strains isolated, 197 strains were identified as A. niger, 131 of which produced FB(2). These strains produced from 1.2 to 27 μg/ml FB(2) in culture. PCR amplification of fum1 and fum19 gene fragments showed that all FB(2)-producing strains and nearly all nonproducing strains of A. niger contain these genes. An additional 175 strains were identified as A. tubingensis, none of which produced FB(2). PCR with fum1 and fum19 primers amplified gene fragments of 14 and 25% of A. tubingensis strains, respectively, suggesting that putative orthologs of A. niger fumonisin biosynthetic genes might occur in A. tubingensis. These results indicate that FB(2) production is common among field isolates of A. niger and suggest that the potential for FB(2) contamination of California raisins should be addressed further.

Between 2004 and 2007 we examined foods from Japanese retail shops for contamination with ochratoxin A (OTA) and fumonisins B(1), B(2), and B(3). A total of 1,358 samples of 27 different products were examined for OTA, and 831 samples of 16 different products were examined for fumonisins. The limits of quantification ranged from 0.01 to 0.5 microg/kg for OTA and 2 to 10 microg/kg for the fumonisins. OTA was detected in amounts higher than limits of quantification in wheat flour, pasta, oatmeal, rye, buckwheat flour and dried buckwheat noodles, raisins, wine, beer, coffee beans and coffee products, chocolate, cocoa, and coriander. OTA was found in more than 90% of the samples of instant coffee and cocoa, and the highest concentration of OTA, 12.5 microg/kg, was detected in raisins. The concentration of OTA in oatmeal, rye, raisins, wine, and roasted coffee beans varied remarkably from year to year. Fumonisins were detected in frozen and canned corn, popcorn grain, corn grits, cornflakes, corn soups, corn snacks, beer, soybeans, millet, and asparagus. The highest concentrations of fumonisins B(1), B(2), and B(3) were detected in corn grits (1,670, 597, and 281 microg/kg, respectively). All of the samples of corn grits were contaminated with fumonisins, and more than 80% of the samples of popcorn grain and corn snacks contained fumonisins. OTA and fumonisins were detected in several food products in Japan; however, although Japan has not set regulatory levels for these mycotoxins, their concentrations were relatively low.

Fumonisins are mycotoxins mainly produced by Fusarium verticillioides, which is a major contaminant of corn. However, there are sporadic reports of fumonisin contamination in wheat worldwide. The rice adherent fungus Gibberella fujikuroi is taxonomically closely related to F. verticillioides. Therefore, the potential risk of fumonisin contamination in rice and wheat is significant. Previously, a sensitive detection method utilizing liquid chromatography with tandem electrospray mass spectrometry (LC-ESI-MS-MS) was developed for the determination of fumonisins in brown rice. In the present study, the incidence of fumonisins in brown rice and wheat harvested in Japan was investigated using LC-ESI-MS-MS. Forty-eight rice samples and 47 wheat samples were screened and analyzed for the major B-type fumonisins: fumonisin B1 (FB1) and fumonisin B2 (FB2). About 1 kg of rice or wheat seed was divided into three subsamples, and 10 g from each subsample was used for the analysis. The limits of detection were 0.012 and 0.011 mg/kg for FBt and FB2, respectively, in rice samples and 0.010 and 0.008 mg/kg for FB1 and FB2, respectively, in wheat samples. The mean (standard deviation) recoveries of FB1 spiked at 0.50 mg/kg into toxin-free rice and wheat samples were 77.6 (4.2)% and 84.5 (3.1)%, respectively. One of the wheat samples was positive for FBt with a value greater than the limit of detection,but no fumonisin was found in any of the rice samples. This is the first report of fumonisins detected in Japanese wheat.

To survey fumonisins B1 (FB1) and B2 (FB2) in agricultural products consumed in South Korea and provide an exposure assessment, ground samples were extracted (80% MeOH), filtered (0.2 microm), and cleaned up. After evaporation, dry residues were reconstituted in 50% MeOH, and a 50-micro1 aliquot of this sample was mixed with 200 micro1 of o-phthaldialdehyde for derivatization. The derivatives were analyzed with a high-performance liquid chromatography system equipped with a fluorescence detector. For validation of the detection procedure, linearity, accuracy, precision, detection limit, and quantification limit were determined. The validated detection method was then used to survey fumonisins in white rice, brown rice, barley, barley tea, beer, wheat flour, millet, dried corn, corn flour, corn tea, canned corn, popcorn, and breakfast cereal. Retention times for FB1 and FB2 standards were 7 and 18 min, respectively. Linearity (R2 = 0.99995 to 0.99998), accuracy (81.47 to 108.83%), precision (2.35 to 5.77), detection limit (25 ng/g or ng/ml), and quantification limit (37 ng/g or ng/ml) indicated that this procedure is capable of quantifying fumonisins in agricultural products. Only FB1-positive samples (5.12%, three dried corn samples and five corn flour samples) were found at 90.89 to 439.67 ng/g. According the survey results, an estimated daily intake of FB1 and FB2 in Korea was 0.087 ng/kg of body weight per day. These results indicate that continuous monitoring of these mycotoxins is necessary to establish appropriate risk assessment, and the maximum tolerable daily intake of fumonisins in Korea is lower than the 2 microg/kg set by the Joint Food and Agriculture Organization-World Health Organization Expert Committee.

Dried figs from Turkey that were visibly moldy (or fluorescent under UV light) and thus rejected as unsuitable for human food were screened for the presence of fungal metabolites. Crude solvent extracts from individual figs were directly analyzed by liquid chromatography combined with time-of-flight mass spectrometry to generate accurate mass data for all detectable components. A comparison of these data with a metabolite database indicated the presence of fumonisins B2 and B4, patulin, HT-2 toxin, and zearalenone among various other metabolites. Portions of the same figs were reextracted and then analyzed by conventional liquid chromatography-mass spectrometry. On the basis of coincident retention times and by matching selected ion monitoring for coincident ions with that of authentic standards, the identification of fumonisin B2, HT-2 toxin, patulin, and zearalenone was confirmed.

Sixty samples of corn from both conventional and organic farms were tested for internal fungal contamination. Molds were identified to genus, and those belonging to the genus Fusarium were identified to species. Twenty isolates of Fusarium verticillioides were tested with a high-performance liquid chromatography-naphthalene dicarboxaldehyde-fluorescence method for their ability to produce fumonisins B1 and B2. The internal fungal infection in organic maize (63.20%) was significantly higher than that in conventional maize (40.27%) (P < 0.05). However, the distribution of fungal genera indicated a significantly higher prevalence of Fusarium in conventional (34.93%) than in organic (18.15%) maize, making Fusarium the predominant fungus in conventional maize. This difference in mold distribution between organic and conventional maize was attributed to the difference in cultivation system. The dominant Fusarium species in both conventional and organic samples was F. verticillioides. There were no significant differences in the ability of 20 selected isolates of F. verticillioides to produce fumonisins on conventional or organic corn. Up to 13.3% of the conventional corn samples contained fumonisins B1 and B2 at mean concentrations of 43 and 22 ng/g, respectively. Organic corn samples had somewhat lower levels of contamination: 35 ng/g fumonisin B1 and 19 ng/g fumonisin B2 (P > 0.05). The organic farming system, with well-balanced crop rotation, tillage, and compost fertilization, produced corn that was less likely to be contaminated with Fusarium species, although no significant difference in fumonisin concentrations was found between the two types of contaminated corn.

We conducted a survey of aflatoxin B1, B2, G1, and G2, ochratoxin A, and fumonisin B1, B2, and B3 contamination in various foods on the retail market in Japan in 2004 and 2005. The mycotoxins were analyzed by high-performance liquid chromatography, liquid chromatography-mass spectrometry, or high-performance thin-layer chromatography. Aflatoxins were detected in 10 of 21 peanut butter samples; the highest concentration of aflatoxin B1 was 2.59 microg/kg. Aflatoxin contamination was not found in corn products, corn, peanuts, buckwheat flour, dried buckwheat noodles, rice, or sesame oil. Ochratoxin A was detected in oatmeal, wheat flour, rye, buckwheat flour, green coffee beans, roasted coffee beans, raisins, beer, and wine but not in rice or corn products. Ochratoxin A concentrations in contaminated samples were below 0.8 microg/kg. Fumonisins were detected in popcorn, frozen corn, corn flakes, and corn grits. The highest concentrations of fumonisins B1, B2, and B3 in these samples were 354.0, 94.0, and 64.0 microg/kg, respectively.

Fusarium species are fungi that infect maize products worldwide and elaborate mycotoxins, which have been associated with cancer. This study was carried out to investigate the potential of lactic acid bacteria fermentation in reducing mycotoxin concentration and toxicity in maize meal products. Maize meal was spiked separately with fumonisin B1 and zearalenone and then allowed to ferment for 4 days. The potential cytotoxicity of the mycotoxin-spiked fermented extracts was also investigated using the SNO human esophageal carcinoma cell line (the SNO cell line was explanted from a cancer patient, S.N., a 62-year-old Zulu man, in July 1972). A significant decrease (P < 0.05) in the concentration of the two mycotoxins was observed, with a 56 to 67% and a 68 to 75% reduction in the third and fourth days, respectively. The two mycotoxins were not detectable in commercially fermented maize meal (amahewu) samples. After fermentation, mycotoxin-spiked maize meal samples containing lactic acid bacteria culture were comparatively less toxic to SNO cells than were samples without lactic acid bacteria. However, this difference in toxicity was not significant (P > 0.05). These results indicate that lactic acid bacteria fermentation can significantly reduce the concentration of mycotoxins in maize. However, such a reduction may not significantly alter the possible toxic effects of such toxins. The exact mechanism of toxin reduction warrants further investigation.

The two major fumonisin-producing Fusarium species are Fusarium verticillioides and Fusarium proliferatum. The growth and fumonisin production of these two isolates on corn was studied at water activities (a(w)) between 0.860 and 0.975 and at temperatures between 15 and 30 degrees C. Growth rates (g, mm/day) were obtained by linear regression during the linear phase of growth. In general, growth rates for both isolates increased significantly (P < 0.05) with increases in a(w) and temperature. Both fumonisin production and radial growth (mycelial development) for both isolates increased with a(w) at all temperatures investigated, but the effect of temperature on this relationship was not obvious. The effect of temperature on fumonisin production at high a(w) values optimal for growth was only marginal, whereas at lower a(w) values the effect of temperature was more pronounced, with more fumonisin production occurring at temperatures not optimal for growth. The optimum temperature for fumonisin production was between 15 and 25 degrees C. For F. proliferatum, the optimum temperature for growth at all a(w) values, 30 degrees C, resulted in the poorest fumonisin production. For both isolates, the slowest initial rate of fumonisin production was at 15 degrees C, the temperature at which the slowest growth rates were obtained.

This review focuses on the fumonisin-producing Fusarium species and the ecophysiology of these species. The effects of environmental biotic and abiotic factors on germination, growth, and fumonisin B1 production by Fusarium verticillioides and Fusarium proliferatum have been investigated under laboratory, field, and storage conditions. An understanding of the factors involved in production of fumonisins is the first step in preventing accumulation of these toxins.

The purpose of this study was to measure the potential levels of fumonisin B1 (FB1) and fumonisin B2 (FB2) contamination in several herbal teas and medicinal plants that are consumed regularly in Turkey. FB1 and FB2 were detected using high-performance liquid chromatography with fluorescence detection after derivatization with o-phthaldialdehyde. A total of 115 commercially available herbal tea and medicinal plant samples were analyzed. The recoveries in black tea were 86.9+/-8.42% for FB1 and 102+/-6.80% for FB2 spiked with 1 microg/g of each analyte. Similarly, the mean recovery results in lime (linden) for FB1 and FB2 were 85.2+/-9.76% and 78.6+/-5.67%, respectively. The minimum detectable amounts for the o-phthaldialdehyde derivatives of FB1 and FB2 were 0.025 microg/g (1 ng injected) and 0.125 microg/g (5 ng), respectively. FB1 was detected in two samples (0.160 and 1.487 microg/g), and FB2 was detected in none of the samples.

Fusarium verticillioides is considered to be the main source of fumonisins, a group of toxins that contaminate commodities and result in chronic and acute diseases affecting humans and animals. The detection and control of this species is crucial to prevent fumonisins from entering the food chain. The objective of the present research was to develop a specific, sensitive, and robust PCR assay to detect F. verticillioides strains using two pairs of specific primers for F. verticillioides, which have been designed on the basis of the intergenic spacer region of the rDNA units. The first pair of primers was F. verticillioides species specific, whereas the second pair of primers detected fumonisin-producing F. verticillioides strains. This second pair of primers allowed for the discrimination between the major group of F. verticillioides strains, fumonisin-producing strains that are mainly associated with crops, and a minor group of strains, non-fumonisin-producing strains that are associated with bananas. Fifty-four strains of F. verticillioides from different geographical regions and hosts were tested using both sets of primers. Sixteen additional Fusarium species were examined. The specificity of the primer sequences provides the basis for a simple, rapid, accurate, and sensitive detection and identification method of this fungal species that represents a risk for human and animal health.

The aim of this study was to investigate the distribution of fumonisin B1 in various corn milling fractions processed by an industrial plant. Corn kernels and six derived milling fractions (germ, bran, large and small grits, animal feed flour, and flour) were sampled. In addition, in order to evaluate the effect of cooking, samples of polenta were prepared starting from naturally contaminated flour obtained from the industrial processing cycle. The industrial plant worked continuously at a rate of 60 tons per day. Two sublots of 5 tons each were investigated with samples of derived products taken at regular time intervals. Due to a similar heterogeneous distribution of fumonisin B1 with other mycotoxins, such as aflatoxins, the sampling scheme was derived from the European Directive 98/53 for aflatoxins. Both lots of kernels showed fumonisin contamination at 4.54 and 5.09 mg/kg, respectively. Germ, bran, and animal feed flour showed contamination levels, namely 8.92 mg/kg (lot 1) and 9.56 mg/kg (lot 2), 7.08 mg/kg (lot 1) and 8.08 mg/kg (lot 2), and 9.36 mg/kg (lot 1) and 6.86 mg/kg (lot 2) higher than large and small grits and flour (0.39 mg/kg [lot 1] and 0.42 mg/kg [lot 2], 0.60 mg/kg [lot 1] and 1.01 mg/kg [lot 2], and 0.40 mg/kg [lot 1] and 0.45 mg/kg [lot 2], respectively). These results seem to account both for the industrial yields of the derived products and the distribution of fumonisin contamination in a kernel. The cooking of polenta in a domestic pressure cooker did not affect fumonisin contamination because the mycotoxin concentrations were similar to those of the starting flour (0.40 and 0.45 mg/kg).

Some species of Fusarium can produce mycotoxins during food processing procedures that facilitate fungal growth, such as the malting of barley. The objectives of this study were to develop a 5' fluorogenic (Taqman) real-time PCR assay for group-specific detection of trichothecene- and fumonisin-producing Fusarium spp. and to identify Fusarium graminearum and Fusarium verticillioides in field-collected barley and corn samples. Primers and probes were designed from genes involved in mycotoxin biosynthesis (TRI6 and FUM1), and for a genus-specific internal positive control, primers and a probe were designed from Fusarium rDNA sequences. Real-time PCR conditions were optimized for amplification of the three products in a single reaction format. The specificity of the assay was confirmed by testing 9 Fusarium spp. and 33 non-Fusarium fungal species. With serial dilutions of purified genomic DNA from F. verticillioides, F. graminearum, or both as the template, the detection limit of the assay was 5 pg of genomic DNA per reaction. The three products were detectable over four orders of magnitude of template concentration (5 pg to 5 ng of genomic DNA per reaction); at 50 ng template per reaction, only the TRI6 and FUM1 PCR products were detected. Barley and corn samples were evaluated for the presence of Fusarium spp. with traditional microbiological methods and with the real-time PCR assay. The 20 barley samples and 1 corn sample that contained F. graminearum by traditional methods of analysis tested positive for the TRI6 and internal transcribed spacer (ITS) PCR products. The five corn samples that tested positive for F. verticillioides by traditional methods also were positive for the FUMI and ITS PCR products. These results indicate that the described multiplex real-time PCR assay provides sensitive and accurate differential detection of fumonisin- and trichothecene-producing groups of Fusarium spp. in complex matrices.

The persistence of fumonisins during cooking is known to be affected by several factors, including thermal degradation and the presence of various ingredients in corn-based food recipes that can react with the toxin. A method for the production of corn kernels containing 14C-fumonisins was developed. The corn kernels were colonized by Fusarium verticillioides MRC 826 and supplemented with 1,2-14C-sodium acetate. The specific activity of 14C-FB1 produced made the study of its fate in cornmeal muffins possible. The double-extraction acetonitrile-water-methanol/immunoaffinity column/o-phthaldialdehyde high-performance liquid chromatography (HPLC) method was used to determine FB1 levels in cornmeal muffins. Reductions in FB1 levels in muffins spiked with 14C-labeled and unlabeled FB1 (43 and 48%, respectively) were similar, indicating that the extraction method was efficient and consistent with previous reports. However, with the labeled corn kernel material, recovery levels based on the 14C counts for the eluate from an immunoaffinity column were much higher (90%). This finding indicates that some fumonisin-related compounds other than FB1 that were present in the cornmeal were recognized by the antibodies but not by the HPLC method.