Natural co-occurrence of aflatoxin and cyclopiazonic acid in peanuts grown in Argentina

Natural occurrence of aflatoxins and cyclopiazonic acid (CPA) contamination in peanuts was investigated. Co-occurrence of CPA and aflatoxins was detected in two of 50 samples analysed. The levels of these toxins found in positive samples were 4300 and 493 μg kg -1 for CPA, 625 and 435 μg kg -1 for aflatoxin B (AFB 1 ), and 625 and 83 μg kg -1 for aflatoxin G (AFG 1 ), respectively. Levels of CPA contamination in the positive samples were similar to those registered in other substrates. This is the first report of natural co-occurrence of CPA and aflatoxins in Argentina.     

Cyclopiazonic acid and aflatoxin production by cultures of Aspergillus flavus isolated from dried beans and maize

From the storerooms of individual households 150 samples of dried beans and 90 samples of stored maize were collected for mycological analyses. Two of 27 isolates of A. flavus grown on malt extract agar (MEA) were found to produce the mycotoxin cyclopiazonic acid (25–36 μg/g). Three of the A. flavus isolates grown on crushed moist wheat produced aflatoxin B₁ (0.72–1.6 μg/g) and 6 of 26 A. ochraceus isolates were OA positive (0.5–10.4 μg/g). None of 25 bean samples were contaminated with CPA, AF or OA, while 4 samples of 30 tested maize samples were OA positive with level of OA 0.4–400 μg/g. Toxins were determined by thin layer chromatography and colorimetric method was used for quantitations of CPA.     

Production of cyclopiazonic acid by aflatoxigenic and non-aflatoxigenic strains of Aspergillus flavus

96 strains of Aspergillus flavus isolated from samples of stored grain and smoke-dried meat products were examined for ability to produce cyclopiazonic acid and aflatoxins, grown on mycological broth medium and malt extract agar. Five strains produced cyclopiazonic acid in the range of 0.5 — 30 mg/kg and 9 produced aflatoxin B1 (0.1 — 14.8 mg/kg) but none of them produced both cyclopiazonic acid and aflatoxins.     

Phenylpyrrole-resistance and aflatoxin production in Aspergillus parasiticus Speare

Mutants of Aspergillus parasiticus highly resistant to phenylpyrroles were isolated at a high mutation frequency, after UV-mutagenesis and selection on media containing fludioxonil. Studies on the effect of mutation(s) on the aflatoxin production resulted in the identification of two fludioxonil-resistant phenotypes: aflatoxigenic (FLD(afl)(+)) and non-aflatoxigenic (FLD(afl)(-)) mutant strains. Most of the FLD(afl)(+) mutant strains produced the aflatoxin B(1) at similar or even higher (up to 2.5-fold) concentrations than the wild-type parent strain on yeast extract sucrose medium. Interestingly, in most of these mutant strains the aflatoxigenic ability significantly increased (up to 4-fold) when the mutants were grown on fungicide-amended medium. However, a significant reduction in the aflatoxin production was observed in wheat grains by all FLD(afl)(+) mutant strains. Tests on the response of mutant strains to high osmotic pressure showed that most fludioxonil-resistant mutants were more sensitive to high osmolarity than the wild-type parent strain. Study of other fitness determining parameters showed that the mutation(s) for resistance to phenylpyrroles may or may not affect the mycelial growth rate, sporulation and conidial germination. However, in a number of aflatoxigenic-mutant strains these fitness parameters were unaffected or only slightly affected. Cross resistance studies with fungicides from different chemical groups showed that the mutation(s) for resistance to fludioxonil also highly reduced the sensitivity of mutant strains to the aromatic hydrocarbon and dicarboximide fungicides. No effect of phenylpyrroles resistance mutation(s) on fungitoxicity of triazoles, benzimidazoles, anilinopyrimidines, phenylpyridinamines, strobilurin-type fungicides and to the non site-specific inhibitors chlorothalonil and maneb was observed. The above mentioned data indicate, for the first time, the potential risk of increased aflatoxin contamination of agricultural products by the appearance and predominance of highly aflatoxigenic mutant strains of A. parasiticus resistant to aromatic hydrocarbon, dicarboximide and phenylpyrrole fungicides.     

Aspergillus flavus and aflatoxin production in acid-treated maize

Isobutyric acid (IBA) and propionic-acetic acid (PA) were applied to comparable 52.8 m³ lots of freshly harvested yellow dent maize containing 27% moisture. After 6 months storage, 30% Aspergillus flavus infection and low levels of aflatoxin were detected in adjacent bins of IBA-treated and PA-treated maize. Extensive samples were taken after 7 months from moldy spots in each bin and evaluated for aflatoxin, zearalenone, ochratoxin and microorganisms. Aspergillus flavus (10⁶ propagules/g) was detected in 40% of the PA samples, but no aflatoxin was found. Also, counts of Aspergillus fumigatus, Absidia and Penicillia were high. In addition to the molds found on PA maize, Aspergillus niger was identified on IBA-treated maize. Aspergillus flavus (10⁴–10⁷ propagules/g) was present in 79% of the IBA samples; aflatoxin (from 2 to 857 ng/g) was detected in 57%. Aflatoxin contamination varied between locations within a moldy area. Among 20 individual kernels picked at random from each location, aflatoxin contamination ranged from 150 to 21.800 ng/g in positive kernels. Evidently, bulk quantities of maize must be appraised on the basis of individual kernels because toxin-free kernels often are adjacent to highly contaminated kernels.     

我国土榨花生油黄曲霉毒素B1 及圆弧偶氮酸毒素污染调查

为了解我国土榨花生油中黄曲霉毒素B1(AFB1)和圆弧偶氮酸(CPA)的污染状况,采集全国12省20市小作坊生产的土榨花生油126份,购买市售品牌花生油27份,采用高效液相色谱法检测花生油中AFB1和CPA两种真菌毒素的含量。结果表明:27份品牌花生油未检出AFB1和CPA; 126份土榨花生油中,AFB1检出率为44.44%,超标率为11.11%,CPA检出率为7.14%,其中河南、福建、广西和山东土榨花生油AFB1污染较为严重。我国土榨花生油存在真菌毒素污染安全隐患,监管部门应严格把控原料及加工环节,确保土榨花生油的质量和安全。     

反相高效液相色谱法分析米曲霉发酵液中的圆弧偶氮酸

为建立米曲霉培养液中圆弧偶氮酸的检测方法 ,采用 4 %乙酸水溶液 (三氟乙酸调至pH2 2 )与乙腈配比做流动相 (5 5 +45 ) ,选择 2 80nm紫外吸收波长 ,测定米曲霉发酵液中的圆弧偶氮酸 ,圆弧偶氮酸在C18色谱柱上获得良好的分离 ,检出限为 0 0 0 2 μg mL ;麦芽汁 -蛋白胨培养液、麦芽汁 -酵母膏培养液、马铃薯 -酵母膏培养液中的圆弧偶氮酸回收率分别为 93 4 %、95 2 %、96 7% ;RSD分别为 4 3%、3 4 %、3 2 %。该方法适宜米曲霉发酵液中圆弧偶氮酸的检测。     

Glufosinate-ammonium reduces growth and aflatoxin B1 production by Aspergillus flavus

The herbicide glufosinate-ammonium (GA) [butanoic acid, 2-amino-4-(hydroxymethylphosphinyl)-ammonium salt] was tested at concentrations from 2 to 2,000 g GA per ml for activity against growth and aflatoxin B1 (AFB) production by the mycotoxigenic fungus Aspergillus flavus Link:Fr. The highest concentration (2,000 microg GA per ml) reduced colony diameter of A. flavus strain AF13 by 80%. AFB1 production was inhibited by 90% at this concentration. Reduction in mycelial dry weight and AFB1 production in response to GA application ranged from 17.2 to 97.1% and from 39.1 to 90.1%, respectively. Of four concentrations tested, 2 microg GA per ml was weakly inhibitory. In the kernel screening assay, AFB1 production was inhibited 60 to 91% when kernels were preimmersed or immersed 5 days after incubation in 200 microg GA per ml. Both concentrations (2 and 200 microg GA per ml) reduced seed germination by 25 to 50%. Results indicate that GA has an inhibitory effect on growth and AFB1 production by A. flavus.     

Inhibition of growth and aflatoxin production of Aspergillus parasiticus by citrus oils

Summary Aspergillus parasiticus was inoculated into grapefruit juice and a glucose-yeast extract medium; both contained 500–7000 ppm of citrus oils that were incorporated into the media by sonication. Orange and lemon oil were more inhibitory to mold growth and aflatoxin production than was d-limonene, the main constituent of the two peel oils. After 7 days at 28° C, 2000 ppm of lemon and 3000 ppm of orange oil in grapefruit juice afforded maximum suppression of mold growth and toxin formation. When the glucose-yeast extract medium was used, 3000 ppm of either oil were needed to achieve the same result. After 4 days at 28° C, orange oil at 3500 ppm in either medium markedly inhibited mold growth (as evidenced by dry weight of mold mycelium) and aflatoxin production (only 14 and 1% of the amount normally produced in the juice and artificial medium, respectively). Higher concentrations of orange oil further reduced mold growth and aflatoxin production and also delayed the onset of sporulation, if it occurred. Although aflatoxin was detected in all samples, only 0.2 to 0.5% of the amount found in controls (without the citrus oil) was present when the medium contained 7000 ppm orange oil. The mold consistently grew, albeit very poorly, on the glass at the liquid-atmosphere interface even when the substrate contained a large amount of citrus oil.

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Hemmung des Wachstums und der Aflatoxinproduktion von Aspergillus parasiticus durch Orangenöl, Citronenöl und d-Limonen

Zusammenfassung Pampelmusensaft und ein Glucose-Hefeextrakt-Medium wurden beide mit 500–7000 ppm Orangenöl, Citronenöl oder d-Limonen angereichert und dann mitAspergillus parasiticus beimpft. Wachstum und Aflatoxinproduktion des Pilzes wurden stärker durch die Öle als durch d-Limonen gehemmt, obwohl dieser der Hauptbestandteil der beiden Öle ist. 2000 ppm Citronenöl bzw. 3000 ppm Orangenöl in Pampelmusensaft genügten zur starken Hemmung der Wachstums- and der Aflatoxinproduktion vonA. parasiticus während 7 Tage bei 28° C. Wenn Glucose-Hefeextrakt als Nährboden diente, dann wiesen bei 3000 ppm beide Öle gleiche Hemmung auf. Wenn beide Nahrboden nur 4 Tage bei 28° C gehalten wurden, dann waren 3500 ppm Orangendl notwendig, um Wachstum and Aflatoxinproduktion zu hemmen. Pampelmusensaft mit einem Orangenöl-Gehalt von 3500 ppm enthielt nur 14% der Aflatoxin-Menge des beimpften Saftes ohne Öl. Das Medium mit Glucose, Hefeextrakt und Orangendl hatte nur 1 % des Aflatoxin-Gehaltes der Kontrolle. Höhere Konzentrationen von Orangenöl hemmten noch stärker und verzögerten den Beginn der Konidienbildung. Wenn das Medium 7000 ppm Orangenöl enthielt, dann konnte nur geringes Pilzwachstum und Aflatoxinproduktion (0,2–0,5% der Kontrolle) beobachtet wurden; das minimale Wachstum des Pilzes geschah an der Grenzfläche Nährboden und Atmosphare.

     

Growth and aflatoxin production by Aspergillus parasiticus in a medium at different pH values and with or without pimaricin

A glucose-yeast extract-salt medium containing 0, 5, 7.5, 10, 15 or 20 micrograms pimaricin/ml with an initial pH of 3.5 or 5.5 was inoculated with Aspergillus parasiticus WB 108 and incubated at 15 degrees or 28 degrees C. The pH, weight of mycelium and amount of aflatoxin produced were determined after 3, 7, and 10 days and after 14, 21, and 30 days when incubation was at 28 degrees or 15 degrees C, respectively. Increasing the concentration of pimaricin in the medium with an initial pH of 5.5 decreased the amounts of aflatoxin B1 and G1 produced after 3 days of incubation. When the initial pH of the medium was 3.5, no growth or toxin production occurred after 3 days of incubation in the medium containing 7.5 micrograms or more of pimaricin/ml. The presence of 20 micrograms of pimaricin/ml inhibited growth and toxin production after 7 days of incubation. When cultures were incubated at 15 degrees C, there was a lag phase which extended from 9 to 16 days, and the amounts of aflatoxin produced decreased with an increasing concentration of pimaricin. Pimaricin did not completely inhibit the growth and aflatoxin production by A. parasiticus. Pimaricin, in combination with a low pH, low temperature or 4% or 6% NaCl, initially caused slow mycelial growth and low toxin production, but the mold overcame the inhibitory effects and produced substantial amounts of mycelium and toxin.     

香菇纤维素衍生物抑制黄曲霉菌产毒的研究

黄曲霉毒素具有诱导突变、抑制免疫和致癌作用。控制黄曲霉毒素污染一直是世界性难题,也是近年来的研究热点之一。实验研究了香菇纤维素衍生物GPX抑制黄曲霉菌产毒活性,初步探究了GPX抑制黄曲霉菌产毒的作用机制。结果表明,10、50μg/mL的GPX对黄曲霉菌产毒的抑制率分别达到70%、95%,而100、250、500、750、1000μg/mL的GPX对黄曲霉菌产毒的抑制率均可达到100%,抑制活性极其明显。GPX离体抗氧化能力很低,但是能够缓解黄曲霉菌丝的氧化胁迫。此外GPX促进了黄曲霉细胞内囊泡过早与大液泡的融合,该现象能够抑制黄曲霉毒素的产生。      

Water activity and temperature effects on growth of Aspergillus niger, A. awamori and A. carbonarius isolated from different substrates in Argentina

The objectives of this study were to determine the effect of water activity, temperature, and their interactions on a) mycelial growth rate and b) the lag phase prior to grow of seven isolates of Aspergillus section Nigri isolated from peanuts, maize kernels, dried grapes and coffee cherries from Argentina. Three Aspergillus niger, three A. awamori and one A. carbonarius isolates examined showed optimum a(W) level for growth at 0.97 with optimal temperature of 30 degrees C. for most of the isolates and 25 degrees C for only one (A. awamori RCP176). Minimal a(W) for growth was 0.85 at the highest temperature tested. Overall growth was reduced up to 50% at 0.93 a(W). Growth was also to a large extend inhibited at 0.85 a(W) for most isolates even after 21 days of incubation at temperatures lower than 30 degrees C. The analysis of variance of the effect of single (isolate, a(W) and temperature), two- and three-way interaction showed that all factors alone and all interactions were statistically significant (P<0.001) in relation to growth rates and lag phase for A. niger, A. awamori and A. carbonarius isolates. These data are relevant since these species are isolated in high frequency on numerous substrates for human and animal consumption in Argentina.     

Updated profile of aflatoxin and Aspergillus section Flavi contamination in rice and its byproducts from the Philippines.

Our goal was to develop an updated profile of aflatoxin (AF) and AF-producing fungi contamination in rice and its byproducts from the Philippines. The total AF levels in 78 samples of polished and brown rice, determined by an immunoaffinity column clean-up method coupled with HPLC (detection limit: 25 ng/kg), ranged from <0.025-2.7 microg/kg (mean of positive samples: 0.37 microg/kg) and 0.03-8.7 microg/kg (mean of positive samples: 2.7 microg/kg), respectively. The incidence (% of positive samples) of AF in polished and brown rice were 94% and 100%, respectively. The AF levels in polished rice imported from Thailand and Vietnam were approximately 20% of the levels found in locally produced polished rice. AF levels decreased as the rice progressed through the various stages in milling. Fungi recovered include toxigenic Aspergillus flavus and A. parasiticus with an incidence ranging from 14% in rice bran to 78% in rough rice and producing <0.025-6200 microg/kg total AF in in vitro cultures on rice. All samples of rice bran and rice hull contained AF at levels ranging from 0.27-11 microg/kg. The estimated potential daily intake of AFB(1) from rice is between 0.1 and 7.5 ng/kg of body weight/day, the mean of which is 1.0 ng representing 9.1-5.3 times the estimated tolerable daily intake for AFB(1) reported to date for Asia. Thus, Filipinos have a potentially high risk of exposure to AF that can be easily controlled through proper post-harvest handling and storage of rice and its byproducts.     

Mycoflora and natural occurrence of aflatoxin, cyclopiazonic acid, fumonisin and ochratoxin A in dried figs

Mycoflora, the mycotoxigenic properties of moulds, and natural contamination with mycotoxins such as aflatoxins (AFs), cyclopiazonic acid (CPA), fumonisin B(1) (FB(1)) and ochratoxin A (OTA) were investigated in dried figs. Dry fig samples were collected from orchards during the drying stage in the Aegean Region of Turkey. Fungal isolates were identified using morphological, chemical as well as molecular methods. Mycotoxigenic characteristics of moulds were assessed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Mycotoxins except CPA (by TLC) were determined by HPLC. All the fig samples were contaminated with moulds and 94.7% contained one or more mycotoxigenic species. The most prevalent moulds present in dried figs belong to the Aspergillus section Nigri members, being 93.9% positive for the samples, followed by Fusarium spp., Aspergillus section Flavi and Penicillium spp. On the other hand, Fusarium spp. had the highest count and the number of fumonisin producing Fusarium was also high. A total of 48% of 115 dried fig samples contained OTA (range = 0.1-15.3 ng g(-1)), 74.7% of the samples had FB(1) (range = 0.05-3.65 mg kg(-1)), 10.0% of the samples had aflatoxin (range = 0.1-763.2 ng g(-1)) and 24.3% of the samples were tentatively identified as being contaminated with CPA (range = 25-187 ng g(-1)). Dried fig samples were contaminated with one (33.0%), two (47.0%), three (5.2%) and four mycotoxins (3.5%). A total of 11.3% of dried fig samples were not contaminated with any of the four mycotoxins. To the best of our knowledge, CPA and fumonisin have been found for the first time in dried figs.     

Mycoflora and natural occurrence of aflatoxin,cyclopiazonic acid,fumonisin and ochratoxin A in dried figs

Mycoflora, the mycotoxigenic properties of moulds, and natural contamination with mycotoxins such as aflatoxins (AFs), cyclopiazonic acid (CPA), fumonisin B₁ (FB₁) and ochratoxin A (OTA) were investigated in dried figs. Dry fig samples were collected from orchards during the drying stage in the Aegean Region of Turkey. Fungal isolates were identified using morphological, chemical as well as molecular methods. Mycotoxigenic characteristics of moulds were assessed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Mycotoxins except CPA (by TLC) were determined by HPLC. All the fig samples were contaminated with moulds and 94.7% contained one or more mycotoxigenic species. The most prevalent moulds present in dried figs belong to the Aspergillus section Nigri members, being 93.9% positive for the samples, followed by Fusarium spp., Aspergillus section Flavi and Penicillium spp. On the other hand, Fusarium spp. had the highest count and the number of fumonisin producing Fusarium was also high. A total of 48% of 115 dried fig samples contained OTA (range?=?0.1–15.3?ng?g^?1), 74.7% of the samples had FB₁ (range?=?0.05–3.65?mg?kg^?1), 10.0% of the samples had aflatoxin (range?=?0.1–763.2?ng?g^?1) and 24.3% of the samples were tentatively identified as being contaminated with CPA (range?=?25–187?ng?g^?1). Dried fig samples were contaminated with one (33.0%), two (47.0%), three (5.2%) and four mycotoxins (3.5%). A total of 11.3% of dried fig samples were not contaminated with any of the four mycotoxins. To the best of our knowledge, CPA and fumonisin have been found for the first time in dried figs.     

Production of cyclopiazonic acid by molds isolated from Taleggio cheese.

Twenty-seven strains of Penicillium were isolated from the rind of Taleggio, a typical Italian cheese, so that we could test their capacity to produce cyclopiazonic acid (CPA); all strains produced CPA. The production was strongly influenced by the strain variety and growth conditions. Strains incubated at 25 degrees C for 7 days always produced CPA in mannitol broth, with concentrations ranging from 0.02 to 1 microg/ml, whereas only 33% of strains grown in yeast-extract broth produced CPA, with a maximum value of 0.1 microg/ml. In milk, maximum production (1.6 microg/ml) was observed after 14 days of incubation at 25 degrees C. In order to evaluate the presence of the toxin and its capacity for migrating into the cheeses, the rind, the cheese near the rind, and the cores from six Taleggio cheeses were analyzed. CPA was present in five cheeses, with a maximum concentration of 0.25 mg/kg in one rind, and in one cheese, the toxin migrated to the core. A positive correlation between CPA production and surface mold was found.     

植物挥发油对黄曲霉毒素产生菌及其毒素合成的影响

黄曲霉毒素(AFT)是由黄曲霉(Aspergillus flavus)和寄生曲霉(Aspergillus parasiticus)等曲霉属真菌产生的次生代谢产物,具有致畸、致癌、致突变等作用.AFT污染多种粮食作物及食品,其极强的毒性严重威胁到人体健康.寻找各种黄曲霉毒素抑制剂已成为国内外研究的热点.部分抗生素类药物及化学杀菌剂虽然具有较强的抗黄曲霉作用,但鉴于其毒副作用较明显,使用范围非常有限.近二十年来,人们开始从天然产物中寻找毒副作用低、易降解并且安全性高的黄曲霉抑制剂.本文对天然植物挥发油及其单体成分抑制黄曲霉和寄生曲霉生长并抑制AFT合成,以及在粮食、食品及其包装等方面的应用情况进行综述,为进一步研究植物挥发油类成分的抑菌机制和产品开发提供参考.