Influence of light on food relevant fungi with emphasis on ochratoxin producing species

The influence of light of varying wavelength on growth and ochratoxin A biosynthesis of Aspergillus carbonarius, A. niger, A. steynii and on Penicillium nordicum and P. verrucosum was analysed. For comparison the influence of light on various other food relevant fungi, including citrinin producers, was also tested. Generally the Aspergilli seem to be more resistant to light treatment than the Penicillia. Interestingly wavelengths from both sides of the spectrum, e. g. red (long wavelength, 627 nm) and blue (short wavelength 470-455 nm) had the strongest inhibitory effects on growth and ochratoxin A biosynthesis. Blue light generally had a stronger effect. Light of moderate wavelength, 590 to 530 nm, (yellow to green) had more a positive than a negative influence on growth or ochratoxin A biosynthesis compared to the control (dark incubation). The light effect on growth and ochratoxin A biosynthesis was dependent on the growth medium. In contrast to malt extract medium (MEA), YES medium, as an especially nutrient rich medium, had an attenuating effect on the reactivity against light. However the tendency of the response in both media was the same. Moreover, the light intensity strongly influences how the fungus reacts. Depending on the intensity and the resistance of the species a complete cessation of growth and/or inhibition of ochratoxin A biosynthesis could be achieved. Light irradiation has the opposite effect on ochratoxin A than citrinin, two mycotoxins which can be produced simultaneously in P. verrucosum. Citrinin was produced essentially under light conditions which inhibited ochratoxin A biosynthesis. The same was true for a derivative of ochratoxin, in particular a derivative of ochratoxin β in A. carbonarius. A. carbonarius produced high amounts of the ochratoxin β derivative under blue light when the production of ochratoxin A was ceased at the most inhibiting conditions used (MEA, royal blue light, 455 nm, 1700 lx). Light has a growth stalling but not inactivating effect on aerial mycelia. If a non-growing colony under light is shifted to the dark it immediately grows normally. However on spores blue light has a deactivating effect. After incubation of spores of P. verrucosum for 24 h under blue light up to 97% of the spores were no longer able to germinate. Again the spores of the Aspergilli were much more resistant.     

Occurrence of the C-series fumonisins in maize from the former Transkei region of South Africa

Fumonisins are a group of structurally related mycotoxins produced mainly in maize by Fusarium verticillioides and F. proliferatum. The most abundant naturally occurring analogue is fumonisin B(1) (FB(1)), with lesser amounts of fumonisin B(2) (FB(2)) and fumonisin B(3) (FB(3)) occurring. The C-series fumonisins (FCs) are structurally analogous to the B-series but lack the C-1 methyl group. Good and mouldy subsistence-grown maize samples were collected from the Centane and Bizana districts in the former Transkei region of South Africa. After extraction with methanol/water and clean-up on strong anion exchange solid phase extraction cartridges, FB(1), FB(2), FB(3), FC(1), FC(3) and FC(4) were determined by reversed-phase LC-MS/MS using positive ion electrospray ionisation. FB(1) levels in both good and mouldy maize from Centane (means (±SD) 2.75 ± 2.24 and 23.4 ± 12.5 mg kg(-1), respectively) were higher than the corresponding levels in maize samples from Bizana (means 0.056 ± 0.157 and 3.71 ± 5.01 mg kg(-1), respectively). Similarly, FC(1) levels in both good and mouldy maize from Centane (means 0.107 ± 0.099 and 0.814 ± 0.391 mg kg(-1), respectively) were higher than in Bizana, where FC(1) was detected in only one (0.018 mg kg(-1)) of 19 good maize samples and occurred in mouldy maize with a mean of 0.102 ± 0.135 mg kg(-1). A significant correlation (r=0.982, p<0.01) was observed between FB(1) and FC(1) levels in all samples, with FC(1) levels at 3.3% of the corresponding FB(1) levels. FC(4) levels were similar to FC(1), whereas only low amounts of FC(3) were detected.     

Occurrence of fumonisins B1 and B2 in broa, typical Portuguese maize bread

Fumonisin B(1) (FB(1)) and fumonisin B(2) (FB(2)) are mycotoxins mainly produced by Fusarium verticillioides, and Fusarium proliferatum, fungi species most commonly isolated from maize. The natural occurrence of FB(1) and FB(2) in broa, typical Portuguese maize bread, was evaluated in 30 samples. Twenty five were found positive with levels ranging from 142 to 550 microg kg(-1). The limit established by the European regulations was exceeded by 27% of the samples. The tolerable daily intake for fumonisin B(1), and B(2), alone or in combination, for all of the analysed samples, was lower than 2 microg kg(-1) body weight per day established by the European Commission.     

Fumonisin B(1) in maize harvested in Iran during 1999.

The fumonisin B(1) (FB(1)) contamination of maize collected in two areas of Iran during 1999 was determined. The 20 maize samples from Mazandaran Province, situated on the Caspian littoral of Iran, consisted of random samples of farmers' lots and were all contaminated with FB(1) at a mean level of 3.18 mg kg(-1) (range 0.68-7.66 mg kg(-1)). The 10 samples (of the same maize cultivar) from Isfahan Province in central Iran were purchased as maize cobs in local retail markets and had mean FB levels of 0.22 mg kg(-1) (mean of all samples, 6/10 samples positive, range <0.01-0.88 mg kg(-1)). The FB levels in Mazandaran, an area of high oesophageal cancer, were significantly (p < 0.0001) higher than the FB levels found in maize from Isfahan, an area of low oesophageal cancer in Iran.     

Two-dimensional profiles of fumonisin B1 production by Fusarium moniliforme and Fusarium proliferatum in relation to environmental factors and potential for modelling toxin formation in maize grain

This study has examined in detail the effect of temperature (7-37 degrees C) and water availability (water activity, a(w), 0.89-0.97) on fumonisin B1 (FB1) production by an isolate of Fusarium moniliforme and F. proliferatum on irradiated maize grain after incubation for 28 days. The optimum conditions for F. moniliforme and F. proliferatum were 30 degrees C at 0.97 a(w) and 15 degrees C at 0.97 a(w), respectively. The maximum concentrations were 2861 mg kg(-1) and 17,628 mg kg(-1) dry wt. maize grain, respectively. At marginal a(w)/temperature conditions for growth (e.g. 0.89-0.91 a(w)) no FB1 was detected (<0.1 mg kg(-1)). A high variability was found between replicates for F. moniliforme, but not for F. proliferatum. These data were used to construct two-dimensional diagrams of all the a(w) x temperature conditions favourable for FB1 production for the first time. The data were also subjected to a polynomical regression, which demonstrated that there was a very good fit for the 15-30 degrees C range of temperature and at 0.97 a(w). However, at marginal environmental conditions this was not possible. This suggests that it may be possible to predict within a limited environmental range the potential for significant FB1 production.     

Survey of the mycobiota of Spanish malting barley and evaluation of the mycotoxin producing potential of species of Alternaria, Aspergillus and Fusarium

The present work deals with the toxigenic mycobiota occurring in Spanish malting barley and the capability for producing mycotoxins by several important toxigenic fungi. One hundred and eighty seven samples of malting barley were gathered from Spanish breweries before processing. One hundred and fifty kernels per sample were surface-sanitized with a 2% sodium hypochlorite solution and incubated on three culture media. The most abundant fungi were species of Alternaria, Aspergillus, Penicillium and Fusarium, which were present in 93%, 82.3%, 57.8% and 27.8% of the samples, respectively. To evaluate their mycotoxin producing potential a number of isolates belonging to each genus, except Penicillium, were randomly selected and incubated on culture media known to be appropriate for production of mycotoxins. Alternariol and alternariol monomethyl ether were produced by 26.7% of Alternaria spp. isolates (all belonged to Alternaria alternata). All tested isolates of F. verticillioides produced fumonisin B(1) (FB(1)) and 61.3% of them produced fumonisin B(2) (FB(2)), whereas FB(1) was synthesized by 83.3% and FB(2) by 77.8% of F. proliferatum isolates. Twenty percent of the isolates of the Aspergillus flavus/A. parasiticus group had the capability to produce aflatoxin B(1) and aflatoxin B(2). Thirty out of 34 isolates of F. graminearum produced deoxynivalenol and zearalenone whereas the other 4 isolates produced nivalenol. Ochratoxin A was detected in 75% and 15% of isolates of Aspergillus section Nigri and A. ochraceus, respectively. This is the first survey carried out in Spain on the toxigenic mycobiota contaminating malting barley in breweries and the mycotoxin producing capacity of several species. The information obtained is useful for assessing the risk of mycotoxins in beer.     

Comparison of fumonisin B1 biosynthesis in maize germ and degermed kernels by Fusarium verticillioides

Fusarium verticillioides produces a group of mycotoxins known as fumonisins in maize kernels. Fumonisins are associated with a variety of mycotoxicoses in humans and animals; thus, their presence in food is a considerable safety issue. This study addressed fumonisin B1 (FB1) production in two components of the maize kernel, namely the germ tissues and the degermed kernel. Growth of F. verticillioides was similar in colonized germ tissue and degermed kernels, but FB1 production was at least five times higher in degermed maize kernels than in germ tissue. Expression of the fumonisin polyketide synthase gene, FUM1, as measured by beta-glucuronidase (GUS) and Northern blot analysis, followed the same pattern as FB1 production. Also correlated to FB1 was a concomitant drop in pH of the colonized degermed kernels. A time course experiment showed that degermed kernels inoculated with F. verticillioides became acidified over time (from pH 6.4 to 4.7 after 10 days of incubation), whereas colonized germ tissue became alkaline over the same period (from pH 6.5 to 8.5). Because conditions of acidic pH are conducive to FB1 production and alkaline pH is repressive, the observed correlation between the acidification of degermed kernels and the increase in FB1 provides one explanation for the observed differences in FB1 levels.     

Comparison of methods and optimisation of the analysis of fumonisins B₁ and B₂ in masa flour, an alkaline cooked corn product

A comparison study of different extraction and clean-up procedures for the liquid chromatographic analysis of fumonisins B(1) (FB(1)) and B(2) (FB(2)) in corn masa flour was performed. The procedures included extraction (heat or room temperature) with acidic conditions or EDTA-containing solvents, and clean-up by immunoaffinity or C18 solid-phase extraction columns. Thereafter an analytical method was optimised using extraction with an acidic mixture of methanol-acetonitrile-citrate/phosphate buffer, clean-up through the immunoaffinity column and determination of fumonisins by liquid chromatography with automated pre-column derivatisation with o-phthaldialdehyde reagent. Recovery experiments performed on yellow, white and blue masa flours at spiking levels of 400, 800 and 1200 μg kg(-1) FB(1) and of 100, 200 and 300 μg kg(-1) FB(2) gave overall mean recoveries of 99% (±6%) for FB(1) and 88% (±6%) for FB(2). Good recoveries (higher than 90% for both FB(1) and FB(2)) were also obtained with corn tortilla chips. The limits of quantification of the method (signal-to-noise ratio of 10) were 25 μg kg(-1) for FB(1) and 17 μg kg(-1) for FB(2). The method was tested on different commercial corn masa flours as well as on white and yellow corn tortilla chips, showing fumonisin contamination levels (FB(1) + FB(2)) up to 1800 μg kg(-1) (FB(1) + FB(2)) in masa flour and 960 μg kg(-1) in tortilla chips. Over 30% of masa flours originating from Mexico exceeded the European Union maximum permitted level.     

伏马毒素的危害及防控技术研究进展

伏马毒素(Fumonisin,FB)是一种产毒真菌的次级代谢产物,广泛存在于玉米等农作物中,严重危害人畜健康。近年来伏马毒素已成为全球污染最严重的霉菌毒素之一,对其的预防和控制已成为研究热点。本文重点介绍了伏马毒素的特性及危害,并对其预防控制技术的研究进展进行了综述,其中生物防治农作物及产品中伏马毒素的方法由于高效且环境友好,具有潜在的应用前景。     

Fumonisin production on irradiated corn kernels: effect of inoculum size.

Production of fumonisins B1, B2, and B3 by Fusarium moniliforme was evaluated on irradiated corn kernels inoculated with different spore concentrations (10, 10(2), 10(3), 10(5), and 10(6)), a water activity of 0.97, and a temperature of 25 degrees C. There was a direct relationship between the level of toxin produced and inoculum size. The highest levels of total fumonisin produced after 35 days of incubation were 5,028 and 9,063 ng/g at 10(5) and 10(6) spores per ml, respectively. The pattern of fumonisin production (FB1 > FB2 > FB3) in cultures growing from different inocula was not affected during the 35 days of incubation. The ratio between FB2 and FB1 varied from 0.15 to 0.42, whereas the ratio between FB3 and FB1 varied from 0.34 to 0.87.     

Fumonsin B1, B2, and B3 content of commercial unprocessed maize imported into South Africa from Argentina and the USA during 1992.

The widespread occurrence of F. moniliforme and the toxic effects of its secondary metabolites, the fumonisins B1(FB1), B2(FB2) and B3(FB3), make it imperative that fumonisin contamination of maize, a major constituent of animal feed as well as the staple diet of many populations, be closely monitored to reduce the risk of fumonisin exposure. Equine leukoencephalomalacia and porcine pulmonary oedema have been associated with the intake of feed heavily contaminated with fumonisins. In addition, high levels of fumonisins in the maize-based staple diets of certain populations have been linked to a high incidence of oesophageal cancer in the Transkei region of South Africa and in Linxian and Cixian Counties, China. Bulk shipments of maize imported into South Africa from the USA and Argentina during 1992 were sampled at the port of entry to determine fumonisin levels. Of the 79 samples from two US shipments, all were positive for fumonisins, with FB1 constituting approximately 71% of the total fumonisins with an overall mean of 2.35 micrograms/gFB1. The maximum FB1 level observed was 3.9 micrograms/g. These levels contrast with those obtained from two Argentinian bulk shipments, which also were all positive for fumonisins, but had a mean FB1 level of 0.31 microgram/g and a maximum observed level of 0.7 microgram/g FB1 measured over 47 composite samples.     

不同光照条件对啤酒花中黄腐酚异构化影响

目的研究不同光照条件,如光照强度、光照时间、处理光的波长等因素对啤酒花中黄腐酚(xanthohumol,Xn)光异构化的影响。方法啤酒花样品在不同光照强度、光照时间、波长和不同颜色玻璃瓶储藏条件下处理后,于OD_(370)处测量Xn含量。结果相同功率光源的紫外光引起Xn异构化较白光显著(P0.05),白光照射Xn样品随着光照时间和光照强度的增大,Xn光异构化程度提高;相同功率光源及光照时间条件下,红光[(625±5)nm]、黄光[(590±5) nm]、绿光[(555±5)nm]、蓝光[(450±5)nm]处理引起的Xn异构化速度差异极显著(P0.01),其中蓝光最强,其次是绿光、红光、黄光;白光光源光照条件下,深棕色玻璃瓶储存对Xn的稳定性显著优于绿色、蓝色、无色玻璃瓶等(P0.05),并且深棕色玻璃瓶长期储存Xn显著优于绿色玻璃瓶(P0.05)。结论光照时间、波长及储存条件对Xn光异构化有显著的影响,且避光储存利于Xn稳定,这为含有Xn的饮品、食品、保健品等的光照或包装条件的选择提供了依据。     

A survey of fumonisins, deoxynivalenol, zearalenone and aflatoxins contamination in corn-based food products in Argentina.

The presence of mycotoxins in corn-based foods available in Argentina was determined in order to make a preliminary exposure assessment. Thirty-eight samples [corn meal ('polenta') and corn flakes] of different local brands were analysed for zearalenone, deoxynivalenol and aflatoxins by TLC and fumonisins (FB1, FB2 and FB3) by HPLC. None of the 38 samples contained any detectable amount of aflatoxins (< 2 micrograms/kg), zearalenone (< 50 micrograms/kg) and deoxynivalenol (< 50 micrograms/kg). By contrast fumonisin contamination was found in 95% of the samples. The highest fumonisin levels were found in corn meal: FB1 (range positives: 60-2860 micrograms/kg; mean positive value: 556 micrograms/kg), FB2 (61-1090 micrograms/kg; 232 micrograms/kg) and FB3 (18-1015 micrograms/kg; 150 micrograms/kg). Low levels of fumonisin B1 were detected in 16/17 corn flakes samples (2-38 micrograms/kg). Total fumonisin levels in corn meal were more than 1000 micrograms/kg in 24% (5/21) of the samples. Although it is not the staple food in Argentina, maize consumption is very important, especially among children. A daily fumonisin intake of 11.3 micrograms/kg of body weight was estimated for child consumers (1-5 years old) based on an average consumption of 200 g of corn meal/day. Calculated at an average rate for all children (consumers or not) the intake estimate was 0.9 microgram/kg of body weight.     

Potential use of antioxidants for control of growth and fumonisin production by Fusarium verticillioides and Fusarium proliferatum on whole maize grain

The effect of interactions between two food grade antioxidants butylated hydroxyanisole (BHA) and propyl paraben (PP, 100, 200, 500 microg g(-1)) and water activity (a(w), 0.995, 0.98, 0.95) of irradiated maize on lag phase prior to growth, growth rate and fumonisin production by Fusarium verticillioides and Fusarium proliferatum was evaluated at 25 degrees C. Both antioxidants had an effect on growth characteristics, and fumonisin production. However, this was dependent on the dose used and the a(w) treatment. At 500 microg g(-1) BHA and PP increased the lag phase prior to growth, and reduced the growth rate of both Fusarium species significantly, especially at 0.95 a(w). Both antioxidants significantly reduced the production of fumonisin by both Fusarium species, especially at 0.98 and 0.95 a(w). These results suggest that these antioxidants have potential for treatment of maize grain for controlling growth of these mycotoxigenic species and prevent fumonisin accumulation.     

基于磁性纳米颗粒伏马菌素B双探针竞争ELISA检测方法的建立

建立一种基于磁性纳米颗粒能同时检测样品中伏马菌素B1(fumonisin B1,FB1)、FB2和FB3总量的双探针竞争酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)检测方法。采用通过活化酯法制备磁性纳米颗粒-FB1捕获探针,利用改良过碘酸钠法合成McAb-HRP探针,对检测步骤进行优化,确定最佳反应条件。所建立检测方法对FB1和FBs的检测范围分别为0.07～1.98 ng/mL和0.10～9.86 ng/mL。在玉米样品中加标回收率范围为86.2%～105.1%。所建立方法检测结果与液相色谱-串联质谱检测结果的相关系数(R²)为0.996 6。双探针竞争ELISA检测法检测速度快、准确性高、重复性好,为玉米中伏FBs的快速筛选提供了新方法。     

Determination of the mycotoxin fumonisin B1 in maize by reversed-phase thin-layer chromatography: a collaborative study

A simple and cost-effective method using thin-layer chromatography for the determination of the mycotoxin fumonisin B₁ in maize is described. The analytical method consisted of the extraction of ground maize by shaking with methanol/water (75:25) for 60 min and clean-up of the resultant extract by means of strong anion exchange solid-phase extraction. The purified residue, formed by evaporation of the elution solvent, was reacted with fluorescamine and the fumonisin B₁-derivative was separated by reversed-phase thin-layer chromatography using a developing solution of methanol/aqueous 4% potassium chloride (70:30). The derivatized FB₁ was readily visualized as a greenish-yellow spot under long wavelength ultraviolet light and quantified by visual comparison with a set of similarly derivatized standards in the range 20-300 ng FB₁ spotted on plate. Based on visual comparison, levels down to 0.5 mg kg^-1 were successfully estimated. The method was collaboratively studied in 14 laboratories using four duplicate maize meal samples (including a blank) and a spiked sample for determination of recovery. No significant difference was observed between mean FB₁ levels by high-performance liquid chromatography or thin-layer chromatography. Based on within-laboratory relative standard deviations of 27.1-41.7% and between-laboratory relative standard deviations of 35.0-63.3%, the method can be considered semiquantitative. The mean recovery achieved by participants at a spiking level of 2.00 mg kg^-1 was 74.5%.     

Local post-harvest practices associated with aflatoxin and fumonisin contamination of maize in three agro ecological zones of Tanzania

A survey was undertaken of a total of 120 farmers, 40 from each of the three studied agro-ecological zones of Tanzania, to determine local post-harvest management practices associated with aflatoxin (AF) and fumonisin (FB) contamination of maize. Data on practices (collected using a structured questionnaire) and maize samples were obtained from each of the 120 farmers. FB and AF contamination in the samples were analysed by HPLC. A total of 45% and 85% of maize samples were positive for AF and FB respectively, with levels ranging from 0.1 to 269 μg kg^?1 for AF and from 49 to 18 273 μg kg^?1 for FBs. Significant differences in contamination level were observed among the three agro-ecological zones. Farmers in the three agro-ecological zones practised similar practices in varying degrees. Drying, sorting and protecting maize against insect infestation are practices that showed significant association with AF or FB contamination of maize. Drying maize on mat/raised platform, sorting (damaged, discoloured and moulded grains) and application of synthetic insecticides during storage are practices that were associated with less contamination of maize with AF and FB. The results can be used to advise on effective post-harvest strategies for prevention of AF and FB contamination of maize in rural Tanzania.     

Occurrence of fumonisin B1 in maize imported into Taiwan

Samples of maize imported into Taiwan during 1997-1998 were collected and analyzed for the presence of fumonisin B1 (FB1) using high performance liquid chromatography. Eight (6.8%) of 118 samples were found to contain FB1 (334-1614 microg kg(-1)). The frequency of FB1 found in maize samples imported from Australia was 20%, followed by Thailand (10%), and USA (5.1%). In analyzing the distribution pattern, it was found that 93.2% of the samples had FB1 concentrations below 100 microg kg(-1), and only 3.4% (or four samples) were in excess of 300 microg kg(-1).     

Formation of ethyl carbamate in umeshu (plum liqueur)

Samples of umeshu, a Japanese plum liqueur made from unripe plums, shochu and crystal sugar, were stored under fluorescent light, in the dark and in the refrigerator. The amount of ethyl carbamate formed in umeshu exposed to light or room temperature was larger than that in the dark or at low temperature. The amount of ethyl carbamate formed in umeshu to which cyanide had been added was larger than that in the absence of added cyanide. Thus, the amount of ethyl carbamate formed in the umeshu was increased by not only light and higher temperature, but also cyanide. Samples of model alcoholic beverages were stored under various conditions using red, yellow and blue cellophanes. The amount of ethyl carbamate formed in the model alcoholic beverage with blue cellophane was larger than in the cases of red and yellow cellophanes. It was found that the amount of ethyl carbamate formed in the model alcoholic beverage was increased by light in the wavelength range of 375-475 nm.     

Absorption, distribution and elimination of fumonisin B(1) metabolites in weaned piglets

The absorption, distribution and elimination of fumonisin B(1) (and B(2)) after oral administration of Fusarium verticillioides (MRC 826) fungal culture, mixed into the experimental feed for 10 days, was studied in weaned barrows. In order to determine the absorption of FB(1) from the feed marked by chromium oxide, a special T-cannula was implanted into the distal part of pigs' ileum. During the feeding of toxin-containing diet (45 mg FB(1) kg(-1)) and until the tenth day after the end of treatment, the total quantity of urine and faeces was collected and their toxin content analysed. At the end of the trial, samples of lung, liver, kidney, brain, muscle, and fat were also collected and their fumonisin content analysed by LC-MS. The fumonisins appeared to decrease the reduced glutathione content in blood plasma and red blood cell haemolysate, possibly associated with in vivo lipid peroxidation. From a data set of 80 individual data and the concentration and rate of C(r) and fumonisins (FB(1), partially hydrolysed FB(1) and aminopentol) in the chymus, it could be established that the accumulative absorption of fumonisin B(1) was 3.9% +/- 0.7%. In the chymus, the FB(1) conversions into aminopentol and partially hydrolysed FB(1) were 1.0 and 3.9%, respectively. The degree of metabolism in faeces was variable, although the main product was the partially hydrolysed form, with very small amounts of the aminopentol moiety being recovered. In the investigated tissues the FB(1) conversion to aminopentol and partially hydrolysed FB(1) was 30 and 20%, respectively.