Mycotoxins in corn-based food products consumed in Brazil: an exposure assessment for fumonisins

Samples from 10 different corn-based food products commercially sold in the Federal District of Brazil were analyzed for fumonisins (FB1 and FB2) using HPLC/fluorescence following naphthalene-2,3 dicarboxaldehyde (NDA) derivatization (limit of quantification (LOQ) = 0.020 mg/kg). Samples were also analyzed for aflatoxins (B1, B2, G1, and G2) on a thin-layer chromatrography (TLC) plate under UV light (LOQ of 2 microg/kg). From the 208 samples analyzed, 80.7 and 71.6% had quantifiable levels of FB1 and FB2, respectively. Mean levels of total fumonisins (FB1 + FB2) ranged from 0.127 mg/kg for corn flakes to 2.04 mg/kg for cornmeal ( creme de milho). No FBs were detected in any of the fresh, sweet corn on the cob samples analyzed. Aflatoxins were not detected in any of the 101 samples analyzed. The daily intakes of fumonisins through the consumption of corn-based food products was estimated using consumption data estimated from the 2002/2003 Brazilian Household Budget Survey and the level of fumonisins found in this and other studies conducted in Brazil. In the Federal District, the calculated total daily intake for the total and the consumers-only populations represented, respectively, 9.0 and 159% of the provisional maximum total daily intake (PMTDI) of 2 microg/kg body weight per day. At the national level, the intakes were calculated based on the fumonisin levels found in the Federal District and on published data from studies conducted elsewhere in the country. They represented 24.1 and 355% PMTDI for the total and the consumers-only populations, respectively. The high incidence of fumonisins in some corn-based products and the exposure levels found for specific subpopulations in the present study indicate the need for setting safe regulatory levels for fumonisins in food in Brazil.     

Surveillance of fumonisins in maize-based feeds and cereals from spain.

A survey has been carried out to determine the levels of fumonisins in 171 samples of maize-based feeds and cereals available in Spain. Also, the samples were examined for mold count and fungal species. Aspergillus, Penicillium, and Fusarium were the most frequent genera, and Fusarium and Aphanocladium had the highest individual percentage counts. Regarding Fusarium species, F. moniliforme (47. 4%) was the predominant species; F. proliferatum (5.3%) and F. subglutinans (7.0%) were isolated at low frequency. The high-performance liquid chromatography-o-phthaldialdehyde fluorescence method was used for the analysis of fumonisins. The highest levels of fumonisins were detected in maize. Overall, fumonisin B(1) (FB(1)) and fumonisin B(2) (FB(2)) were detected in 79.5 and 14.6%, of samples respectively, with average FB(1) levels of 3.3 microg/g and average FB(2) levels of 1.7 microg/g. Low levels of fumonisins in wheat, barley, and soybean were detected. This would appear to be the first report of concentrations of fumonisins in these commodities.     

Toxicological assessment of recombinant xylanase X(22) in wine

Toxicological evaluation of xylanase X(22) from Aspergillus nidulans expressed in a wine yeast strain was carried out. The safety of the X(22) intake was assessed by digestibility, bioinformatic, and mouse short-term repeated dosing studies, although X(22) shows resistance to proteolytic degradation in the gastrointestinal system, is a minority protein component (<0.5 10(-)(6) %) of the produced wine, and shows no significant amino acid sequence homology to any known food allergens. The 4-week oral toxicity study was performed in Swiss mice at a dose level of 0.01, 0.1, or 1 mg/kg/day (these dosages correlate to 8, 80, and 800 times, respectively, the enzyme amount contained in 250 mL of wine). Body weight, food and fluid intake, urinalysis, and hematology data were obtained. Postmortem examinations and histopathology by both light and electron microscopy were performed. According to the results of this study, no adverse effects were detected by oral administration of X(22).     

Determination of N-(carboxymethyl)fumonisin B(1) in corn products by liquid chromatography/electrospray ionization--mass spectrometry

It is well-known that fumonisin B(1) (FB(1)) in corn meal decreases during baking, frying, and cooking, but it is still not exactly clear how heating affects the formation of N-(carboxymethyl)fumonisin B(1) (NCM-FB(1)), the reaction product of FB(1) and reducing sugars. In model experiments corn grits were spiked with FB(1) (2 mg/kg) and D-glucose (50 g/kg) or sucrose (50 g/kg) and manufactured into extrusion products at various temperatures (160--180 degrees C) and moisture levels (16--20%). A liquid chromatography/electrospray ionization-mass spectrometry method using isotopically labeled fumonisin FB(1)-d(6) as an internal standard was developed for the determination of NCM-FB(1). For sample cleanup solid-phase C18 cartridges were used. The detection limit achieved with this method was 10 ng/g (signal-noise ratio = 3:1) using the protonated molecule [M + H](+) signal of NCM-FB(1) (m/z 780) in the selected ion monitoring mode. Low concentrations of NCM-FB(1) (29-97 ng/g) were detected in all samples spiked with D-glucose and FB(1), whereas those spiked with FB(1) and sucrose showed only NCM-FB(1) in samples produced at 180 degrees C (NCM-FB(1) = 27 ng/g). Various corn-containing food samples from the German market were analyzed for the presence of NCM-FB(1), FB(1), and hydrolyzed fumonisin B(1) (HFB(1)). All samples were contaminated with FB(1) (22--194 ng/g) and HFB(1) (5--247 ng/g). Six of nine samples contained NCM-FB(1) in low concentrations ranging from 10 to 76 ng/g. From these data and the low toxicity of NCM-FB(1) it can be concluded that the significance of NCM-FB(1) in food seems to be a minor one.     

Occurrence of fumonisins B(2) and B(4) in retail raisins

Concerns that raisins may be contaminated by fumonisins stem from the persistent occurrence of Aspergillus niger spores on raisins and the recent discovery of fumonisin production by A. niger on grapes, which leads to the widespread occurrence of fumonisin B(2) in wine. This study presents an LC-MS/MS survey of fumonisins in retail raisins. In 10 of 21 brands collected in Denmark, Germany, and The Netherlands, fumonisins B(2) and B(4) were detected at levels up to 13 and 1.3 μg/kg, respectively. Only fumonisin B(2) has been detected in wine, so the presence of fumonisin B(4) in raisins suggests that the fumonisins are produced mainly during the drying process concomitant with the decreasing water activity. Analysis of multiple packages from one manufacturer showed a 3-fold package-to-package variation, suggesting that a few raisins per package are contaminated.     

Evaluation of fumonisin contamination in cornflakes on the Belgian market by "flow-through" assay screening and LC-MS/MS analyses

A total of 205 cornflake samples collected in Belgian retail stores during 2003-2004 were surveyed for the natural occurrence of fumonisin B1 (FB1), B2 (FB2), and B3 (FB3). These cornflake samples, originating from conventional as well as from organic production, were analyzed using an intralaboratory-validated LC-MS/MS method. Additionally, 90 cornflake samples were subjected to rapid screening using a flow-through enzyme immunoassay method to demonstrate the practicability of a screening test coupled to a validated confirmatory LC-MS/MS method for the management of food safety risks. FB(1) concentrations ranged from not detected (nd) [LOD (FB1) = 20 microg/kg] to 464 microg/kg with mean and median concentrations of respectively 104 +/- 113 and 54 microg/kg. For FB2 and FB3, the concentration ranges varied respectively from nd [LOD (FB2) = 7.5 microg/kg] to 43 microg/kg and from nd [LOD (FB3) = 12.5 microg/kg] to 90 microg/kg. Mean concentrations for FB2 and FB3 were respectively 12 +/- 8 and 21 +/- 15 microg/kg, while the median concentration was 11 microg/kg for FB2 and 19 microg/kg for FB3. From the statistical tests (chi2 and ANOVA model III), it could be concluded that the agricultural practice did not have any significant effect on the fumonisin concentrations but that the variation between different batches was significant (p < 0.0001).     

Natural occurrence of fumonisins B(1) and B(2) in domestic maize of Taiwan.

Samples of maize grown in various districts of Taiwan were collected and analyzed for the presence of fumonisin B(1) (FB(1)) and fumonisin B(2) (FB(2)) using high-performance liquid chromatography. Forty-nine (44.5%) and 2 (1.8%) of 110 samples were found to contain FB(1) (109-1148 ng/g) and FB(2) (222-255 ng/g), respectively. The frequency of detection and also the maximum FB(1) concentration were found in samples from Penton (2/2, 262 ng/g), followed by Chiayi (18/26, 264 ng/g), Tainan (8/16, 160 ng/g), Hualinen (5/14, 1148 ng/g), Taitung (7/20, 109 ng/g), and Yunlin (9/26, 361 ng/g). Of the 110 samples examined, only 2 samples from Hualinen had been detected containing FB(2). During an analysis of the distribution pattern of FB(1), it became apparent that >79% of tested samples had FB(1) concentrations <100 ng/g, whereas 2.7% (or 3 samples) contained FB(1) >300 ng/g. These results clearly illustrated that domestically produced maize for human consumption is frequently contaminated with FB(1).     

Occurrence of ochratoxin A- and aflatoxin B1-producing fungi in Lebanese grapes and ochratoxin a content in musts and finished wines during 2004

This paper reports the results of an extensive survey on the occurrence of filamentous fungi isolated from wine-grapes in Lebanon and to test their ability to produce ochratoxin A (OTA) and aflatoxin B1 (AFB1) on CYA culture medium, in order to assess their potential for producing these mycotoxins on grapes. From the 470 grapes samples taken during season 2004, 550 fungi strains were isolated with 490 belonging to Aspergillus spp. and 60 belonging to Penicillium spp. All these isolated fungi starins were tested for their ability to produce OTA and AFB1. Aspergillus carbonarius shows that it is the only species able to produce OTA with a production percentage reaching 100% and a maximum concentration of 52.8 microg/g of Czapek yeast extract agar (CYA). In its turn, Aspergillus flavus was considered as the only AFB1-producing species with production percentage of 45.3% and a maximum concentration reaching 40 microg/g CYA. A total of 47 handmade musts produced from the collected grapes were also analyzed in order to correlate the presence of OTA in must and the occurrence of filamentous fungi on grapes; 57.4% were contaminated with OTA at low level with concentrations ranging between 0.011 and 0.221 microg OTA L(-1). The analysis of these must samples was not performed with regard to AFB1. Seventy samples of finish red wine were also assayed for OTA content. The results showed that 42 of the tested samples (60%) were found to be positive for OTA with low levels (0.012-0.126 microg OTA L(-1)).     

Reaction of fumonisin with glucose prevents promotion of hepatocarcinogenesis in female F344/N rats while maintaining normal hepatic sphinganine/sphingosine ratios

The reaction of the primary amine of fumonisin B(1) (FB(1)) with glucose was hypothesized to detoxify this mycotoxin. Eighty 10-day-old female F344/N rats were injected intraperitoneally with diethylnitrosamine (DEN; 15 mg/kg of body weight). At 4 weeks of age, the weaned rats were randomly assigned to one of four treatment groups with 20 rats each. At 9 weeks of age, four rats from each treatment group were killed. At 12 weeks, another five rats from each group were killed. At 20 weeks of age, the remaining rats were killed. In comparison with the rats fed basal diet or FB(1)-glucose (containing 25 ppm of FB(1)), rats fed 8 ppm (residual amount of free FB(1) in the FB(1)-glucose mixture) or 25 ppm of FB(1) had greater alanine aminotransferase activity at 9 and 20 weeks of age (P < 0.001), greater endogenous hepatic prostaglandin E(2) production at 20 weeks of age (P < 0.05), and significantly lower plasma cholesterol at 20 weeks of age (P < 0.01). Placental glutathione S-transferase (PGST)-positive and gamma-glutamyltransferase (GGT)-positive altered hepatic foci (AHF) occurred only in rats fed 25 ppm of FB(1) at 20 weeks of age. Hepatic natural killer (NK) cell activities were similar among the four groups, but the percentage of total liver-associated mononuclear cells exhibiting the NKR-P1(bright) marker was significantly greater in rats fed FB(1)-glucose, FB(1) (8 ppm) and FB(1) (25 ppm) than in control rats at 9 weeks of age, and FB(1)-glucose-treated rats had significantly lower NKR-P1(bright) cells as a percentage of total liver-associated mononuclear cells than rats fed 25 ppm of FB(1) at 20 weeks of age (P < 0.05). PGST- or GGT-positive AHF were not detected in any treatment group at 9 or 12 weeks of age. At 20 weeks of age, half of the rats fed 25 ppm of FB(1) had PGST- and GGT-positive AHF. The sphinganine (Sa) concentration and the Sa/sphingosine (So) ratio were significantly greater in the rats fed 25 ppm of FB(1) diet as compared with the control groups at, respectively, 12 or 20 weeks of age. Therefore, modifying FB(1) with glucose seems to prevent FB(1)-induced hepatotoxicity and promotion of hepatocarcinogenesis. The Sa/So ratio was not the most sensitive biomarker of FB(1) toxicity.     

Production of the mycotoxins fusaproliferin and beauvericin by South African isolates in the Fusarium section Liseola

The production of fusaproliferin (FUS), a recently described mycotoxin, and beauvericin (BEA), a mycotoxin recently reported to co-occur with FUS in Fusarium-infected corn, by South African isolates in the Fusarium section Liseola, was investigated. Five isolates each of F. verticillioides, F. proliferatum, F. subglutinans, and F. globosum were cultured on corn kernels. Four each of the five South African isolates of F. proliferatum and F. subglutinans produced FUS (10-1725 and 330-2630 mg/kg, respectively). BEA was produced by four of the F. proliferatum strains (310-1130 mg/kg) and three of the F. subglutinans strains (140-700 mg/kg). The isolates of F. verticillioides failed to produce significant levels of either of these secondary metabolites. F. globosum was a weak producer of both in that one isolate of five produced 25 mg/kg FUS and five out of five produced BEA at levels ranging between 10 and 110 mg/kg. To further characterize these strains, their production of fumonisins B(1), B(2), and B(3), as well as moniliformin, was investigated. Of the four species investigated, fumonisins were produced by all except F. subglutinans, which in turn was the only species whose isolates in this study produced moniliformin (four of five isolates, ranging from 155 to 2095 mg/kg). Analysis of visibly Fusarium-infected home-grown corn collected in the Transkei region of the Eastern Cape Province of South Africa showed that nine of the ten samples contained low levels of FUS (up to 62 microg/kg), whereas all ten samples showed BEA contamination ranging from 8 to 1734 microg/kg with a mean of 258 microg/kg.     

Liquid chromatography/mass spectrometry investigation of the impact of thermal processing and storage on peach procyanidins

Normal-phase liquid chromatography/mass spectrometry (LC/MS) was used to determine the levels and fate of procyanidins in frozen and canned Ross clingstone peaches as well as in the syrup used in the canning over a 3 month period. Procyanidin oligomers, monomers through undecamers, were identified in Ross clingstone peaches. Optimized methods allowed for the quantitation of oligomers through octamers. The profile of procyanidins in peaches is similar to profiles found in grapes, chocolate, and beverages linked to health benefits such as tea and wine. The monomer content in frozen peeled peaches was found to be 19.59 mg/kg. Dimers (39.59 mg/kg) and trimers (38.81 mg/kg) constituted the largest percent composition of oligomers in the peaches. Tetramers through octamers were present in levels of 17.81, 12.43, 10.62, 3.94 and 1.75 mg/kg, respectively. Thermal processing resulted in an 11% reduction in monomers, a 9% reduction in dimers, a 12% reduction in trimers, a 6% reduction in tetramers, and a 5% reduction in pentamers. Hexamers and heptamers demonstrated an approximate 30% loss, and octamers were no longer detected. Analysis of the syrup after thermal processing indicates that there is a migration of procyanidin monomers through hexamers into the syrup that can account for the losses observed during the canning process. Storage of canned peaches for 3 months demonstrated a time-related loss in higher oligomers and that by 3 months oligomers larger than tetramers are not observed. At 3 months postcanning, levels of monomers had decreased by 10%, dimers by 16%, trimers by 45%, and tetramers by 80%. A similar trend was observed in the canning syrup.     

Evolution of chlorpyrifos, fenarimol, metalaxyl, penconazole, and vinclozolin in red wines elaborated by carbonic maceration of Monastrell grapes

The influence of the different steps involved in the wine-making process on the disappearance of chlorpyrifos, fenarimol, metalaxyl, penconazole, and vinclozolin in red wines elaborated by carbonic maceration of Monastrell grapes was studied. The initial levels of the residues in grapes ranged from 0.28 mg/kg (penconazole) to 1 mg/kg (chlorpyrifos). Ten days after the beginning of maceration, the compound that had decreased least was chlorpyrifos (83% of initial value remaining), whereas metalaxyl, the least persistent of the residues, had decreased to 49% of its initial value. In the free-run juice, on the other hand, the highest percentage remaining corresponded to metalaxyl (10%) and the lowest to chlorpyrifos (0. 1%). After pressing, the percentages of initial values eliminated in pomace varied from 82.7% for chlorpyrifos to 17.7% for metalaxyl, whereas in the press juice the opposite was the case (37% metalaxyl and 2% chlorpyrifos remaining). In finished wine, there were residues of all the pesticides, with the exception of chlorpyrifos, metalaxyl (21%) being the most persistent. The percentages eliminated in the lees varied from 1.5 to 2.5% of the initial value.     

Residues of spiroxamine in grapes following field application and their fate from vine to wine

Dissipation of the fungicide spiroxamine in grapes of two vine varieties, Roditis and Cabernet Sauvignon, exposed to field treatments was evaluated. Vines of a grape vineyard located in central Greece were sprayed once or twice with a commercial formulation of the fungicide at 30 g a.i./hL. Residues in grapes, must, and wine were determined by gas chromatography/IT-MS after extraction with cyclohexane-dichloromethane (9:1), with a limit of quantitation 0.02 mg/kg in grapes and 0.012 mg/kg in wine. Under field conditions, spiroxamine dissipation on grapes was faster during the first 2 weeks and then slower to the sixth week. About 7 days after application, half of the initial spiroxamine concentration remained on the grapes; the respective proportion at 42 days was about 10%. At 14 and 35 days, residues were lower than 0.44 and 0.22 mg/kg, respectively, values below the maximum residue levels set by the European Union (1 mg/kg). Spiroxamine residues transferred from grapes into the must and through the vinification process into the wine were also studied. Mean transfer factors of 0.26 and 0.55 were found from grapes into wine for the wines obtained without maceration and with maceration, respectively. Residues in wine, prepared from grapes with a spiroxamine content of 0.11-0.20 mg/kg, varied from <0.026 to 0.09 mg/kg. Spiroxamine diastereomer B was found to dissipate slower than diastereomer A in the field as well as during the vinification process.     

水稻秸秆降解复合菌系的筛选构建及其田间应用效果

  【目的】  筛选并构建适宜原位还田水稻秸秆快速腐解的高效多功能复合菌系,以提高秸秆原位还田的腐解速率。  【方法】  秸秆原位腐解菌株从水稻田带有腐烂秸秆的表层土壤中筛选分离,分别采用DNS法、摇瓶培养观察法和失重法测定了腐解菌株的羧甲基纤维素酶活性、滤纸崩解能力及水稻秸秆降解能力,对降解效果较好的菌株进行了16S rRNA或18S rRNA鉴定。选取无拮抗作用的菌株构建复合菌系,并在实验室条件下测定各复合菌系的滤纸酶活性和秸秆降解率。挑选降解效果较好的复合菌系进行田间试验,分析了秸秆降解率、土壤有机质及速效养分含量,调查了水稻产量。  【结果】  从土壤中共分离到秸秆降解率均高于19.8%的菌株有6个,包括B2、B5、B6、F1、F5、F12,经鉴定分别为贝莱斯芽孢杆菌(Bacillus velezensis,B2)、耐盐芽孢杆菌(Bacillus halotolerans,B5)、枯草芽孢杆菌(Bacillus subtilis,B6)、米曲霉(Aspergillus oryzae,F1)、黑曲霉(Aspergillus niger,F5)、长枝木霉(Trichoderma longibrachiatum,F12),其中F12 (长枝木霉)的秸秆降解率最高(29.1%)。以该6个菌株构建了7组复合菌系,复合菌系B2+F12、F1+F5+F12、B2+F5+F12的秸秆降解效果较好,在液体培养基中,对秸秆的降解率分别达到32.9%、29.8%、40.3%。利用这3组复合菌系进行田间试验,结果表明,复合菌系B2+F5+F12表现最好,与未施菌剂相比,原位还田的水稻秸秆降解率提高了37.1个百分点,土壤有机质、碱解氮、速效磷、速效钾含量分别增加了2.1 g/kg、1.9 mg/kg、0.6 mg/kg、1.7 mg/kg,水稻产量提高了6.3% (P<0.05)。  【结论】  施用复合菌系B2+F5+F12不仅具有良好的水稻秸秆降解能力,同时能增加土壤有机质、碱解氮、速效磷、速效钾含量,并促进作物产量的提高,具有较好的应用潜力。     

Production of fumonisin B and C analogues by several fusarium species

Six strains of Fusarium verticillioides, two of F. oxysporum, one strain of F. proliferatum, and a strain of an unidentified species were cultured on maize patties and rice and evaluated for their ability to simultaneously produce fumonisin B (FB) and C (FC) series analogues. Fumonisins were quantified by LC-MS-MS using positive ion electrospray ionization. FC1 provided characteristic fragment ions at m/z 690, 672, 654, 532, 514, and 338 corresponding to sequential loss of H2O and tricarboxylic acid moieties from the alkyl backbone, while FC3 and FC4 provided equivalent product ions 16 and 32 amu lower than the corresponding FC1 fragments, respectively. All isolates cultured on maize produced FC4. All isolates except for that of F. proliferatum also produced FC1, and three of the six strains of F. verticillioides produced FC3. All isolates except those of F. oxysporum produced detectable amounts of FB1, FB2, and FB3. Isolates that produced fumonisin B analogues produced at least 10 fold more of the B series analogues than they did of the C series analogues. The results confirm that at least some strains of F. oxysporum produce FC, but not FB, fumonisin analogues and also suggest that the genetics and physiological regulation of fumonisin production may be more complicated than previously envisaged since some strains of F. verticillioides and F. proliferatum as well as the strain of the unidentified species can simultaneously produce both FB and FC analogues.     

Effect of baking and frying on the in vivo toxicity to rats of cornmeal containing fumonisins

Fumonisins are mycotoxins produced by Fusarium verticillioides (=F. moniliforme) and other Fusarium species. They are found in corn and corn-based foods. Cooking decreases fumonisin concentrations in food products under some conditions; however, little is known about how cooking effects biological activity. Baked cornbread, pan-fried corncakes, and deep-fried fritters were made from cornmeal that was spiked with 5% w/w F. verticillioides culture material (CM). The cooked materials and the uncooked CM-spiked cornmeal were fed to male rats (n = 5/group) for 2 weeks at high (20% w/w spiked cornmeal equivalents) or low (2% w/w spiked cornmeal equivalents) doses. A control group was fed a diet containing 20% w/w unspiked cornmeal. Toxic response to the uncooked CM-spiked cornmeal and the cooked products included decreased body weight gain (high-dose only), decreased kidney weight, and microscopic kidney and liver lesions of the type caused by fumonisins. Fumonisin concentration, as determined by HPLC analysis, in the 20% w/w pan-fried corncake diet [92.2 ppm of fumonisin B(1) (FB(1))] was slightly, but not statistically significantly, lower than those of the 20% w/w baked cornbread (132.2 ppm of FB(1)), deep-fried fritter (120.2 ppm of FB(1)) and CM-spiked cornmeal (130.5 of ppm FB(1)) diets. Therefore, baking and frying had no significant effect on the biological activity or concentration of fumonisins in these corn-based products, and the results provided no evidence for the formation of novel toxins or "hidden" fumonisins during cooking.     

Fumonisin B(1)-nonproducing strains of Fusarium verticillioides cause maize (Zea mays) ear infection and ear rot

Fumonisins are polyketide mycotoxins produced by Fusarium verticillioides (synonym F. moniliforme), a major pathogen of maize (Zea mays) worldwide. Most field strains produce high levels of fumonisin B(1) (FB(1)) and low levels of the less-oxygenated homologues FB(2) and FB(3), but fumonisin B(1)-nonproducing field strains have been obtained by natural variation. To test the role of various fumonisins in pathogenesis on maize under field conditions, one strain producing FB(1), FB(2), and FB(3), one strain producing only FB(2), one strain producing only FB(3), and one fumonisin-nonproducing strain were applied to ears via the silk channel and on seeds at planting. Disease severity on the harvested ears was evaluated by visible symptoms and by weight percent symptomatic kernels. Fumonisin levels in kernels were determined by high-performance liquid chromatography. The presence of the applied FB(1)-nonproducing strains in kernels was determined by analysis of recovered strains for fumonisin production and other traits. All three FB(1)-nonproducing strains were able to infect ears following either silk-channel application or seed application at planting and were as effective as the FB(1)-producing strain in causing ear rot following silk-channel application. These results indicate that production of FB(1), FB(2), or FB(3) is not required for F. verticillioides to cause maize ear infection and ear rot.     

Fumonisin in tortillas produced in small-scale facilities and effect of traditional masa production methods on this mycotoxin

Four small tortilla plants were visited in Cameron County, Texas, where observations were made on their production methods. Samples of liquids and solids were collected at each stage of the nixtamalization process, and the pH was recorded. Samples were analyzed for fumonisin B(1) (FB(1)) using an immunoaffinity column/HPLC method chosen for its sensitivity for FB(1). It was found that production methods were highly variable among the producers visited, with major differences particularly in the amount of lime added and boiling times. As reported by others working in Mexico and Central America, FB(1) was found in some tortillas. This led to studies of the effects of the various recipes and across a greater range of initial FB(1) concentration/damaged corn than has typically been reported. Five initial concentrations of FB(1) were tested using irradiated corn kernels inoculated with Fusarium verticillioides MRC 826 as the source of FB(1). The amount of FB(1) detected in the masa and tortillas decreased as the concentration of Ca(OH)(2) increased, and boiling time had no apparent effect. Unexpectedly, as the initial concentrations were increased in the corn prior to nixtamalization, greater percentage reductions in FB(1) were observed.     

Resistance screening essay of wine lactic acid bacteria on lysozyme: efficacy of lysozyme in unclarified grape musts

In wine making, the bacteriolytic activity of lysozyme has primarily been used to control the malolactic fermentation in wines. The use of lysozyme in musts before settling and the beginning of the alcoholic fermentation to inhibit the growth of lactic acid bacteria could be very beneficial. In a resistance test carried out in MT/b broth, lysozyme had greater antimicrobial activity toward Oenococcus oeni than Lactobacillus species. Several strains of wine bacteria belonging to Oenococcus proved sensitive to the bacteriolytic activity of lysozyme at low concentrations in both synthetic medium (MT/b) (50 mg/L), white must, or red must made with or without the skins (100 mg/L). Lactobacillus and Pediococcus strains survived at lysozyme concentrations of 200-500 and 500 mg/L, respectively, in MT/b and musts. Suspended solids in unclarified musts may strongly bind to lysozyme thereby causing its removal by filtration or centrifugation. One hour after lysozyme was added to musts, it was quantified by HPLC and found after centrifugation to be 40-50% and only 10% in musts made with or without the skins, respectively. Although appreciable amounts of lysozyme were bound to wine components, this did not appear to be a serious hindrance to lysozyme activity.     

Determination of free proline and monosaccharides in wine samples by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD)

A sensitive and selective analytical method for the simultaneous separation and quantitative determination of proline and free monosaccharides in wine samples by high-performance anion-exchange chromatography coupled with pulsed amperometric detection is described. Under optimized experimental conditions, a complete separation was obtained in less than 30 min, using an isocratic elution with 10 mM NaOH and 1 mM Ba(OAc)(2). No postcolumn addition of strong bases to the eluent for enhancing detection sensitivity was needed. Upon 25-fold sample dilution and purification to avoid interference of tannins, pigments, and phenolic compounds, the fingerprinting of common monosaccharides (i.e., arabinose, glucose, fructose, galactose, and xylose) and proline in wines, musts, and vinegars can be easily accomplished. The method allows high recovery and satisfies the necessary requirements for accuracy, repeatability, and sensitivity. Values obtained for proline content ranged from 470 to 1190 mg/L in "Aglianico" red wines (mean value, 870 +/- 192 mg/L, n = 21) and from 168 to 286 mg/L in white wines (mean value, 208 +/- 32 mg/L, n = 11). Lower levels were found in musts of red and white grapes, 550 and 87 mg/L, respectively. The lowest content of proline, ca. 10 mg/L, was found both in white and red vinegars.