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.     

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.     

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.     

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。     

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。     

Effect of Volatile Components of Carrot Seed Oil on Growth and Aflatoxin Production by Aspergillus parasiticus

The effect of the volatile fraction of carrot seed oil (VCSO) and its components on the growth and aflatoxin production by Aspergillus parasiticus was studied. Geraniol, citral and terpineol prevented growth and therefore aflatoxin production. VCSO inhibited growth and no aflatoxin was produced. Limonene and terpinene did not affect growth but inhibited aflatoxin production. The addition of VCSO, limonene and terpinene decreased the rate of primary metabolism as demonstrated by the higher final pH of the medium. VCSO, limonene and terpinene reduced the growth rate, measured by the incorporation of [³H] amino acids into trichloracetic acid (TCA) insoluble protein. The addition of VCSO at anytime up to 5 days reduced the aflatoxin accumulation at 7 days.     

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...     

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).     

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.     

Aflatoxin and Cyclopiazonic Acid Production by Aspergillus flavus Isolated from Contaminated Maize

Seven truck-loads of maize were tested for mycotoxin contamination. Aflatoxin was identified in all 7 at concentrations from 3 ng/g-501 ng/g (aflatoxin B₁+ B₂). Cyclopiazonic acid was identified in 4 loads with concentrations from 25-250 ng/g. Deoxynivalenol was found in 4 of 5 loads tested, over a range of 46-676 ng/g. Ninteeen isolates of Aspergillus flavus from the samples were tested for ability to accumulate cyclopiazonic acid and aflatoxin in liquid culture. Fourteen produced cyclopiazonic acid (0.5-135 μg/mL), 12 produced aflatoxin (0.01-0.70 μg/mL, aflatoxin B₁+ B₂), and one aflatoxin-producing isolate did not produce cyclopiazonic acid.     

Influence of Temperature, pH, Water Activity and Antifungal Agents on Growth of Aspergillus flavus and A. parasiticus

Effect of temperature, pH, water activity, and nine antifungal agents on growth of Aspergillus flavus and A. parasiticus was determined on Sabouraud-Dextrose Agar and on corn. Maximal growth of the two molds occurred at 33°C, the highest temperature used, pH of 5.0 and a_w of 0.99. At 15°C, growth was observed at a_w of 0.95 but not 0.90. Slight growth was observed at an a_w, of 0.85 at 27°C and 33°C. Nine antifungal agents (Botran, Orthocide, Poly-ram 80, Topsin-M, Thiram, Imazalil, sodium propionate, sodium sulfite and DDVP) were tested for inhibition of growth. Activity of the antifungals increased as the a_w was decreased. All antifungals showed inhibitory activity, but Imazalil and DDVP were the most effective agents at the lowest concentrations.     

Influence of Some Spice Essential Oils on Aspergillus Parasiticus Growth and Production of Aflatoxins in a Synthetic Medium

Gas-liquid chromatography was used to determine the essential oil compositions of thyme, cumin, clove, caraway, rosemary, and sage. The basic components of these oils were thymol, cumin aldehyde, eugenol, carvonc, borneol and thujonc, respectively. The antifungal potential of the oils against Aspergillus parasiticus were investigated. The essential oils caused complete inhibition of both mycelial growth and aflatoxin production. The effectiveness followed the sequence: thyme > cumin > clove > caraway > rosemary > sage. The major components of the essential oils produced an inhibitory effect at minimum inhibitory concentrations equal to those obtained with the oils.     

Aflatoxin-producing potential of Aspergillus flavus and Aspergillus parasiticus isolated from samples of smoked-dried meat

Over a period of three years 420 samples of various smoke-dried meat products, collected from individual households in different region of Croatia were analysed for the presence of aflatoxigenic strains of the Aspergillus flavus group. Strains of A. flavus and A. parasiticus were present in 17,8% of the samples, and aflatoxin-producing ability was tested in 75 strains. In relation to sequential method of aflatoxin detection, 5 of 8 isolates were found in the first step (fluorescence in aflatoxin-producing ability medium - APA) and all of them in the second step (extraction method from syntheses on moist shredded wheat - SW). A. flavus strains produced mainly aflatoxin B₁, and had various levels of toxigenicity (1.4–3.12 mg/kg). Some strains of A. parasiticus produced all four aflatoxins B₁ B₂ G₁ G₂, while the other ones produced AF B₁ + G₁ only, with concentrations of aflatoxins from 0.1 to 450 mg/kg.     

Aqueous extracts of Tulbaghia violacea inhibit germination of Aspergillus flavus and Aspergillus parasiticus conidia

Aspergillus flavus and Aspergillus parasiticus are important plant pathogens and causal agents of pre- and postharvest rots of corn, peanuts, and tree nuts. These fungal pathogens cause significant crop losses and produce aflatoxins, which contaminate many food products and contribute to liver cancer worldwide. Aqueous preparations of Tulbaghia violacea (wild garlic) were antifungal and at 10 mg/ml resulted in sustained growth inhibition of greater than 50% for both A. flavus and A. parasiticus. Light microscopy revealed that the plant extract inhibited conidial germination in a dose-dependent manner. When exposed to T. violacea extract concentrations of 10 mg/ml and above, A. parasiticus conidia began germinating earlier and germination was completed before that of A. flavus, indicating that A. parasiticus conidia were more resistant to the antifungal effects of T. violacea than were A. flavus conidia. At a subinhibitory extract dose of 15 mg/ml, hyphae of both fungal species exhibited increased granulation and vesicle formation, possibly due to increased reactivity between hyphal cellular components and T. violacea extract. These hyphal changes were not seen when hyphae were formed in the absence of the extract. Transmission electron microscopy revealed thickening of conidial cell walls in both fungal species when grown in the presence of the plant extract. Cell walls of A. flavus also became considerably thicker than those of A. parasiticus, indicating differential response to the extract. Aqueous preparations of T. violacea can be used as antifungal treatments for the control of A. flavus and A. parasiticus. Because the extract exhibited a more pronounced effect on A. flavus than on A. parasiticus, higher doses may be needed for control of A. parasiticus infections.     

Efficacy of Some Essential Oils Against Aspergillus flavus with Special Reference to Lippia alba Oil an Inhibitor of Fungal Proliferation and Aflatoxin B1 Production in Green Gram Seeds during Storage

During mycofloral analysis of green gram (Vigna radiata (L.) R. Wilczek) seed samples taken from different grocery stores by agar and standard blotter paper methods, 5 fungal species were identified, of which Aspergillus flavus exhibited higher relative frequency (75.20% to 80.60%) and was found to produce aflatoxin B₁. On screening of 11 plant essential oils against this mycotoxigenic fungi, Lippia alba essential oil was found to be most effective and showed absolute inhibition of mycelia growth at 0.28 μL/mL. The oil of L. alba was fungistatic and fungicidal at 0.14 and 0.28 μL/mL, respectively. Oil had broad range of fungitoxicity at its MIC value and was absolutely inhibited the AFB1 production level at 2.0 μL/mL. Chemical analysis of this oil revealed geranial (36.9%) and neral (29.3%) as major components followed by myrcene (18.6%). Application of a dose of 80 μL/0.25 L air of Lippia oil in the storage system significantly inhibited the fungal proliferation and aflatoxin production without affecting the seed germination rate. By the virtue of fungicidal, antiaflatoxigenic nature and potent efficacy in storage food system, L. alba oil can be commercialized as botanical fungicide for the protection of green gram seeds during storage.     

Effects of Onion (Allium cepa L.) and Garlic (Allium sativum L.) Essential Oils on the Aspergillus versicolor Growth and Sterigmatocystin Production

Abstract: In the present study the effects of individual and combined essential oils (EOs) extracted from onion (Allium
cepa L.) bulb and garlic (Allium sativum L.) clove on the growth of Aspergillus versicolor and sterigmatocystin (STC) production were investigated. The EOs obtained by hydrodistillation were analyzed by GC/MS. Twenty one compounds were identified in onion EO. The major components were: dimethyl‐trisulfide (16.64%), methyl‐propyl‐trisulfide (14.21%), dietil‐1,2,4‐tritiolan (3R,5S‐, 3S,5S‐ and 3R,5R‐ isomers) (13.71%), methyl‐(1‐propenyl)‐disulfide (13.14%), and methyl‐(1‐propenyl)‐trisulfide (13.02%). The major components of garlic EO were diallyl‐trisulfide (33.55%), and diallyl‐disulfide (28.05%). The mycelial growth and the STC production were recorded after 7, 14, and 21 d of the A. versicolor growth in Yeast extract sucrose (YES) broth containing different EOs concentrations. Compared to the garlic EO, the onion EO showed a stronger inhibitory effect on the A. versicolor mycelial growth and STC production. After a 21‐d incubation of fungi 0.05 and 0.11 μg/mL of onion EO and 0.11 μg/mL of garlic EO completely inhibited the A. versicolor mycelial growth and mycotoxins biosynthesis. The combination of EOs of onion (75%) and garlic (25%) had a synergistic effect on growth inhibition of A. versicolor and STC production. Practical Application: A substitution of synthetic preservatives with natural antimicrobial compounds in food safety to control fungal contamination and mycotoxin production.     

Inhibition of growth and mycotoxin production of Aspergillus flavus and Aspergillus parasiticus by extracts of Agave species

In this work, the effect of ethanolic, methanolic and aqueous extracts of Agave asperrima and Agave striata on growth and production of aflatoxin (in A&M medium) and cyclopiazonic acid (CPA; in Czpaek-Dox medium) and on growth in corn under storage conditions was determined. Aspergillus strains were inoculated (10(6) conidia per ml of medium or per 6 g of corn), then plant extracts were added and incubated without shaking at 28 degrees C for 8 days (for aflatoxin-producing analysis) or for 12 days (for CPA-producing analysis). Aflatoxin was assayed by HPLC and cyclopiazonic acid by absorbance at 580 nm using the Erlich reagent. The extracts that most effectively inhibited growth were those from the flowers of both plants. These exhibited an MIC from 0.5 to 2 mg/ml in culture media. Extracts from scape showed an MIC from 15 to 30 mg/ml in culture media. The MIC of the flower extracts was higher (>30 mg/g) when examined in corn. However, concentrations lower than the MIC drastically inhibited production of aflatoxins in culture medium or in corn. Half of the MIC inhibited 99% of the production of aflatoxins and 85% of cyclopiazonic acid.     

Effect of Calcium Propionate and Water Activity on Growth and Aflatoxins Production by Aspergillus flavus

ABSTRACT: The efficacy of calcium propionate at 2 different doses (0.5% and 1%) against growth and aflatoxins production by Aspergillus flavus (A-2092) was investigated in vitro on Czapek yeast extract agar at different levels of water activity (a_w) in the range of 0.94 to 0.996a_w. A. flavus spores germinated on all calcium propionate and a_w treatments; however, 1% calcium propionate at 0.94 a_w delayed the germination process for up to 10 d. The growing rate of mycelia was slower (0.28 mm/d) at 1% calcium propionate and 0.94 a_w. Aflatoxins (B1, B2, G1, and G2) were also produced minimally (36.1, 1, 1.86, and 1.01 ng/g of media, respectively) at the aforementioned dose rate of calcium propionate and water activity. It was concluded that addition of calcium propionate and a_w amelioration can prove effective tools for suppressing the germination, growth rate, and aflatoxins production by A. flavus in substrate.