Effect of Soybean Volatile Compounds on Aspergillus flavus Growth and Aflatoxin Production

ABSTRACT:  Soybean homogenates produced volatile compounds upon exposure to lipase. These induced volatiles were identified by SPME. Seventeen volatile compounds identified by SPME were chosen for determination of their ability to inhibit Aspergillus flavus growth and aflatoxin B₁ (AFB1) production in a solid media assay. These volatiles included aldehydes, alcohols, ketones, and furans. Of the tested compounds, the aldehydes showed the greatest inhibition of fungal growth and AFB1 production. These compounds inhibited up to 100% of the observed growth and AFB1 production as compared to the controls. The greatest activity by the aldehydes to disrupt growth was ranked as follows: 2,4 hexadienal > benzaldehyde > 2-octenal > ( E )-2-heptenal > octanal > ( E )-2-hexenal > nonanal > hexanal. The greatest activity by the aldehydes to reduce AFB1 was ranked as follows: ( E )-2-hexenal > 2,4 hexadienal > ( E )-2-heptenal > hexanal > nonanal. ( E )-2-hexenal and ( E )-2-heptenal were tested further in an A. flavus -inoculated corn kernel assay. Both compounds prevented colonization by A. flavus and eliminated AFB1 production when exposed to compound volumes < 10 μL as also shown in the solid media assay. The results suggest that soybeans react to lipase by production of potent antifungal volatiles.     

Effect of Cycling Temperatures on Aflatoxin Production by Aspergillus parasiticus and Aspergillus flavus in Rice and Cheddar Cheese

Initiation of growth, sporulation and aflatoxin production at cycling temperatures took less time than at 15°C but more than at 18°C and 25°C. A. parasiticus produced more aflatoxins on rice under cycling temperatures than at 25°C, 18°C or 15°C, while A. flavus produced less aflatoxin under cycling temperatures. A. parasiticus produced more aflatoxins on cheese under cycling temperatures than at 18°C or 15°C, but much less than at 25°C. A. flavus produced less aflatoxins on cheese under cycling temperatures than at 18°C and 25°C. Both organisms produced trace amounts of toxins at 15°C on cheese. Preincubation at 25°C for 2 days before temperature cycling did not increase aflatoxin production on rice but increased production on cheese. The rate of aflatoxin production on cheese decreased as the temperature decreased. No growth, sporulation or aflatoxin production was observed at 5°C on either rice or cheese.     

Effects of Sorbate and Propionate on Growth and Aflatoxin Production of Sublethally injured Aspergillus parasiticus

Growth and aflatoxin production by injured spores (heat, 55°C for 15 min, γ-irradiation, 50 Krad) of Aspergillus parasiticus in the presence of sorbate (0.05 and 0.1%) and propionate (0.2 and 0.4%) were studied in YES broth at pH 4.5, 5.5 and 6.5, and 25°C for 21 days. Aflatoxin production was accelerated in the early stages of growth by γ-irradiation, but not heat. Growth and aflatoxin were delayed in cultures from low numbers of uninjured spores. Aflatoxin increased and was produced sooner by low numbers of uninjured spores in 0.05% sorbate. Both inhibitors slowed growth of injured spores more than non-injured. Inhibition of aflatoxin was dependent on the concentration of inhibitor, pH and injury.     

Increased Aflatoxin Production by Aspergillus parasiticus Under Conditions of Cycling Temperatures

The effects of cycling temperatures (5°C for 12 hr and 25°C for 12 hr) on aflatoxin production by Aspergillus parasiticus NRRL 2999 in yeast extract sucrose (YES) medium were studied. Cycling temperatures, after preincubation at 25°C for various times, resulted in more aflatoxin B₁, G₁, and total aflatoxin production than did constant incubation at either 25°C, which is generally considered to be the optimum for aflatoxin production, or 15°C, which is the same total thermal input as the 5-25°C temperature cycling. With increased preincubation time at 25°C, toxin production increased and the lag phase of growth was shortened or not evident. Cultures that were preincubated at 25°C for 1, 2, and 3 days prior to onset of temperature cycling showed the greatest increase in maximum aflatoxin production over the 25°C and 15°C constant temperatures. Cultures that were not preincubated at 25°C but subjected to constantly fluctuating temperatures produced maximum amounts of aflatoxin equivalent to cultures incubated at a constant 25°C. The maximum aflatoxin production at all temperatures studied occurred during the late log phase of growth and at pH minimums. Aflatoxins were found in higher concentrations in the broth than the mycelia under temperature cycling conditions, at 15°C, and at 25°C during the first 21 days of incubation, whereas greater amounts of toxin were retained in mycelium at 25°C in the later incubation period (28-42 days).     

Effects of Tropical Citrus Essential Oils on Growth, Aflatoxin Production, and Ultrastructure Alterations of Aspergillus flavus and Aspergillus parasiticus

Ethyl acetate extracts and hydrodistillated essential oils from five cultivars of tropical citrus epicarps were evaluated for their inhibitory activities against Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus, Aspergillus parasiticus, and Penicillium sp. using disk diffusion and broth microdilution assays. Essential oils prepared from kaffir lime (Citrus hystrix DC) and acid lime (Citrus aurantifolia Swingle) epicarps exhibited stronger antifungal activity to all fungi than their ethyl acetate extracts with minimum inhibitory concentration and minimum fungicidal concentration values of 0.56 and 1.13 mg/ml (dry matter), respectively, against aflatoxin-producing A. flavus and A. parasiticus. The dominant components of the essential oil from kaffir lime were limonene, citronellol, linalool, o-cymene, and camphene, whereas limonene and p-cymene were major components of acid lime essential oil. Pure limonene, citronellal, and citronellol were five to six times less fungicidal than the natural essential oils, indicating the synergistic activity of many active compounds present in the oils. Kaffir and acid lime essential oils significantly reduced aflatoxin production of A. flavus and A. parasiticus, particularly lime essential oil, which completely inhibited growth and aflatoxin production of A. flavus at the concentration of 2.25 mg/ml. Target cell damage caused by acid lime essential oil was investigated under transmission electron microscopy. Destructive alterations of plasma and nucleus membrane, loss of cytoplasm, vacuole fusion, and detachment of fibrillar layer were clearly exhibited in essential-oil-treated cells.     

Aspergillus Parasiticus Growth and Aflatoxin Production on Dehydrated Taro

A study was made of Aspergillus parasiticus growth and aflatoxin production on four taro media. The critical equilibrium relative humidity (ERH) for natural mold growth on unsterilized dehydrated taro was 88% at 20°C. However, nontoxigenic A. parasiticus NRRL 1957 did not grow at this ERH on dehydrated raw taro incubated at 20°, 30°, or 40°C. Instead, the growth of A. parasiticus NRRL 1957 on dehydrated taro was optimum at 30°C and an ERH of 96%. Aflatoxin production by toxigenic A. parasiticus NRRL 2999 was investigated on four taro media under optimal growth conditions. Only moderate quantities of aflatoxins were produced by A. parasiticus NRRL 2999 on uncooked dehydrated taro, but cooking or supplementation with peptone stimualted mycelial growth and aflatoxin production slightly. Nevertheless, growth and aflatoxin production on cooked or peptone-supplemented taro media was low.     

Property Characterization of Peanut Kernels Subjected to Gamma Irradiation and Its Effect on the Outgrowth and Aflatoxin Production by Aspergillus parasiticus

Peanut kernels inoculated with Aspergillus parasiticus conidia and uninoculated kernels were gamma irradiated with 0 to 15 kGy using ⁶⁰Co. Levels of 2.5 and 5.0 KGy were effective in retarding the outgrowth of A. parasiticus and reducing the population of natural mold contaminants. However, elimination of these molds was not achieved. When irradiated with doses higher than 10 KGy, seed germinations were inhibited, changes in proteins were observed and oil stabilities decreased. After 4 wk incubation of the inoculated kernels in a humidified condition, aflatoxins produced by surviving A. parasiticus ranged from 69.12 to 13.48 μg/g depending upon the original irradiation dose.     

芝麻油挥发性风味成分的研究

采用水蒸气蒸馏，无水乙醚萃取浓缩，气相色谱质谱联用测定了市售芝麻油的挥发性风味成分，实验共分离鉴定了57种化合物。占总检出化合物的81．66％，检出化合物可分为吡嗪，吡咯，吡啶，噻唑，噻吩，呋喃和酚类。     

Streptomyces alboflavus TD-1产挥发性抑菌物质对黄曲霉菌生长及其毒素的抑制作用

利用双皿对扣和气相色谱-质谱法,研究白黄链霉菌TD-1(Streptomyces alboflavus TD-1)在不同葡萄糖浓度培养基中的生长和挥发性有机化合物(volatile organic compounds,VOCs)代谢规律,通过荧光显微镜、扫描电子显微镜及高效液相色谱等方法,研究1-辛烯-3-醇对黄曲霉菌...     

云南大叶种晒青毛茶提取物对产毒黄曲霉生长及产毒的影响

普洱茶在发酵过程中,微生物组成十分复杂,尤其是黑曲霉等霉菌起到主要作用,因此一些消费者担心普洱茶在发酵过程中受到黄曲霉毒素(AFS)的污染。本文通过测定菌落直径、孢子萌发及菌丝体干重等方法研究云南大叶种茶提取物对产毒黄曲霉AS3.4408生长的影响;采用紫外荧光法和HPLC法研究云南大叶种茶提取物对产毒黄曲霉AFS生物合成的影响;并将产毒黄曲霉AS3.4408接种到云南大叶种茶叶中,检测茶叶中的AFS含量,以求对普洱茶的安全性进行评估。研究表明,云南大叶种茶提取物对产毒黄曲霉AS3.4408菌落的生长及产毒均具有显著的抑制作用,且存在明显的剂量依赖关系;将产毒黄曲霉AS3.4408接种到云南大叶种茶叶中,菌株生长良好,但茶叶基质经HPLC检测,未检测到黄曲霉毒素B1、B2、G1和G2,表明云南大叶种茶中的某种(些)成分对黄曲霉毒素的生物合成具有抑制作用。     

基于GC-IMS 技术分析不同提取方式对辣椒籽油挥发性成分的影响

以四平头辣椒籽为原料,对原料进行120℃、10 min烘烤处理,采用气相离子迁移色谱技术对冷榨法、溶剂法和超声辅助溶剂法3种方式提取的辣椒籽油挥发性成分进行分析,运用主成分分析法明确烘烤处理及不同提取方式对辣椒籽油挥发性物质的影响。结果表明,3种不同提取方式辣椒籽油挥发性物质成分特征明显。在烘烤前后不同提取方式辣椒籽油的特征风味中,未烘烤冷榨法、烘烤冷榨法、未烘烤溶剂法、烘烤溶剂法、未烘烤超声辅助溶剂法、烘烤超声辅助溶剂法中的挥发性成分种类分别为38、38、20、18、11、13种。冷榨法对辣椒籽油挥发性成分保留的最好,其呈香物质主要为醛类、酯类、醇类和酮类。烘烤处理辣椒籽油中未出现新的挥发性成分,但促进异戊醛、正丁醛、3-甲基-2-丁醇、正丙醇、异丁醇等呈香物质的形成。     

河南鹤壁瓜蒌子和瓜蒌皮中挥发油成分分析

采用减压蒸馏的方法提取瓜蒌子和瓜蒌皮中挥发油成分。采用气相色谱-质谱联用法对河南鹤壁瓜蒌子和瓜蒌皮中挥发油的化学成分进行分离和鉴定。从瓜蒌子共鉴定出13种化合物,主要为醛类(66.58%)、脂肪酸类化合物(12.4665%),还含有少量烃类(2.1819%)、醇类(3.922%)、酮类(2.4887%)、苷类(3.1607%)和三萜类化合物(2.1459%)等,其总和占挥发油的92.82%;从瓜蒌皮中鉴定得到26个化合物,主要为醛类(57.09%)、脂肪酸类(15.04%)、烃类(9.78%)、酮类(4.7%)等化合物,其含量之和占挥发油的93.03%。瓜蒌子和瓜蒌皮挥发油中均含(E,E)-2,4-壬二烯醛、2,4-癸二烯醛、(E)-2-庚烯醛、亚油酸、棕榈酸,角鲨烯。研究结果为瓜蒌类药材产品及食品添加剂的开发提供参考。     

不产毒黄曲霉菌对产毒黄曲霉菌产毒抑制效果分析

本实验6株菌分离自广东、山东、辽宁和湖北四省的花生土壤中,通过形态学和分子生物学鉴定均为黄曲霉菌,HPLC测定其产毒能力,其中GZ-6为产毒菌,GZ-15、WF-5、WF-20、JZ-2和YC-8为不产毒菌。分别以花生和玉米为培养基,将不产毒黄曲霉菌和产毒菌(孢子浓度:104:105或105:105)进行混合培养,测定不产毒菌对产毒黄曲霉产毒的抑制效果。结果显示:不产毒菌对产毒菌产毒的抑制率随着其孢子浓度的增加而明显加强,当孢子浓度比为105:105(不产毒菌:产毒菌)时,5株不产毒菌在玉米培养基上对产毒菌产毒的抑制率为34.55%~75.94%,在花生培养基上对产毒菌产毒的抑制率为38.03%~83.03%,其中WF-5、WF-20和GZ-15这三株不产毒菌对产毒黄曲霉产毒的抑制效果均达到75.00%以上,可以作为田间防治黄曲霉毒素污染的候选菌株。     

Growth and biosynthesis of aflatoxin by Aspergillus parasiticus in cultures containing nisin

Nisin, 200 or 5000 Reading units/ml, was added to Aspergillus parasiticus cultures. The cultures were incubated at 28 degrees C for 3, 7 or 10 days and analyzed for mycelial dry weight, pH and accumulation of aflatoxin B1 and G1. During the first 3 days of incubation, dry weight, pH decrease and aflatoxin accumulation were suppressed by nisin, when compared with similar values for the nisin-free control. After longer incubation, differences in dry weight nd pH values decreased, whereas accumulation of aflatoxin in the nisin-containing cultures surpassed that of the control.     

Growth and biosynthesis of aflatoxin by Aspergillus parasiticus in cultures containing nisin

Summary Nisin, 200 or 5000 Reading units/ml, was added toAspergillus parasiticus cultures. The cultures were incubated at 28 °C for 3, 7 or 10 days and analyzed for mycelial dry weight, pH and accumulation of aflatoxin B₁ and G₁. During the first 3 days of incubation, dry weight, pH decrease and aflatoxin accumulation were suppressed by nisin, when compared with similar values for the nisin-free control. After longer incubation, differences in dry weight and pH values decreased, whereas accumulation of aflatoxin in the nisin-containing cultures surpassed that of the control.

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Wachstum und Aflatoxin-Biosynthese von Aspergillus parasiticus in Nisin-enthaltenden Kulturen

Zusammenfassung Nisin (200 oder 5000 Reading-Einheiten/ml) wurdeAspergillus parasiticus Kulturen zugegeben. Die Kulturen wurden bei 28 °C für 3, 7 oder 10 Tage bebrütet und auf Mycel-Trockengewicht, pH und Aflatoxin-Inhalt geprüft. Während der ersten drei Tage wurden das Wachstum und die Aflatoxin-Biosynthese durch Nisin etwas gehemmt, nach längere Incubation jedoch gefördert.

     

山苍子精油抑制黄曲霉菌的生长和产毒作用研究

山苍子精油是一种纯天然植物精油,本文研究了其对黄曲霉生长、代谢和毒素产生的抑制作用,探讨了山苍子精油对黄曲霉菌的抑菌能力和作用机理。本研究将花生放置于自然环境染菌并分离纯化目标菌,采用形态学并结合ITS序列法进行菌株分类鉴定;结合抑菌圈、抑菌率和最低抑菌浓度(MIC)的测定探讨山苍子精油对黄曲霉菌的抑制能力;进行了山苍子精油影响黄曲霉孢子萌发率、生长曲线和黄曲霉毒素B1产生的实验研究;从细胞膜渗透性、细胞酶活性的变化探讨了山苍子精油抑制黄曲霉的作用机理。实验结果表明:从腐败花生中分离筛选出菌株HB₂,经ITS序列法鉴定为黄曲霉(Aspergillus flavus);黄曲霉素测定结果显示其含有黄曲霉素B1(AFB1),质量浓度为3.4×10³μg·kg^-1(纯湿菌体);抑菌圈随精油浓度的增大明显变大,对黄曲霉的最低抑菌体积分数(MIC)为0.800μL·mL^-1;孢子萌发率、牙管长度、黄曲霉菌体的生长量和AFB1的浓度随培养液中精油浓度的增大呈显著下降趋势,当山苍子精油浓度为0.100μL·mL     

吲哚对黄曲霉生长及产毒的抑制作用

本文研究了吲哚对黄曲霉的生长以及产毒的抑制作用。通过96孔板微量法发现吲哚能够抑制黄曲霉的生长,其最小抑制浓度(MIC)为100μg/m L;进而采用差量法测定了吲哚对黄曲霉菌丝生长量的抑制作用,结果表明200μg/m L的吲哚处理可以完全抑制黄曲霉的菌丝生长。此外,通过高效液相色谱分析发现吲哚浓度达到50μg/m L时,尽管对菌丝生长量没有明显影响,但是可以有效地抑制黄曲霉毒素B1的产生。这些结果说明吲哚抑制毒素产生并不是通过抑制生长来实现的。为了进一步了解吲哚对毒素产生的抑制机制,本研究通过RT-PCR分析了吲哚对黄曲霉产毒相关基因表达的影响。结果显示吲哚对产毒调控基因afl R的影响与其对毒素的作用趋势相同,同时吲哚还能够下调其他一些产毒相关基因afl K和alf D的表达。我们的研究结果表明吲哚具有很高的开发价值应用于粮食和饲料中黄曲霉毒素污染的控制。     

精炼过程对油茶籽油挥发性化合物的影响研究

为了解精炼过程对油茶籽油风味的影响,采用顶空固相微萃取和气相色谱质谱联用技术（HS-SPME-GC-MS）分析了精炼各阶段油茶籽油挥发性化合物的变化规律。结果表明,精炼过程油茶籽油中共鉴定出76种挥发性化合物,其中,气味活度值（OAV）> 1的关键风味化合物有37种;毛油、脱胶油、脱酸油、脱色油、脱臭油和脱蜡油的挥发性化合物和关键风味物质分别有56与29、51与30、51和30、54和33、42和25、39与24种;油茶籽油中挥发性化合物主要是由酯类、醛类、醇类、酸类、杂环类、酮类和烯类组成,分别有24、15、13、7、6、5和4种,精炼各阶段挥发性化合物中浓度最高的基本上都是醛类,其次是醇类和酯类;随着精炼过程的进行,油茶籽油中挥发性化合物的种类逐渐减少,尤其是酯、醇和杂环的种类减少明显,由毛油中的18、11、6种化合物减少至脱蜡油中的13、7、1种,挥发性化合物的浓度,醛类、酮类、醇类、酸类和酯类整体上呈现出先上升后下降的趋势。精炼过程对油茶籽油风味的影响较大是脱色、脱臭两个工序,脱色油中酸类物质显著增加,产生发霉、腐败和汗味等不良气味,脱臭后,挥发性化合物的种类和浓度明显降低,风味强度减弱,不利于浓香油茶籽油生产。