Isolation of methyl syringate as a specific aflatoxin production inhibitor from the essential oil of Betula alba and aflatoxin production inhibitory activities of its related compounds

Methyl syringate was isolated from the essential oil of Betula alba as an aflatoxin production inhibitor. It inhibited aflatoxin production of Aspergillus parasiticus and Aspergillus flavus with IC₅₀ values of 0.9 and 0.8 mM, respectively, without significantly inhibiting fungal growth. Methyl syringate reduced mRNA levels of genes (aflR, pksA, and omtA) encoding proteins required for aflatoxin biosynthesis. Methyl gallate, methyl 3,4,5-trimethoxybenzoate, and methyl 3-O-methylgallate inhibited both aflatoxin production and fungal growth of A. parasiticus and A. flavus. However, their acids and syringic acid did not inhibit aflatoxin production and growth of A. parasiticus significantly, although gallic acid inhibited aflatoxin production of A. flavus with selectivity. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of methyl syringate was much weaker than that of gallic acid. These results showed that methyl syringate has a unique inhibitory activity toward aflatoxin production with a different mode of action from that of gallic acid.     

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.     

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.     

Production of aflatoxins in sausage,salami, sucuk and kavurma

Forty nine meat product samples were examined for the fungal genera. Penicillium sp. was detected in 74.8% of samples. No sample contained Aspergillus parasiticus or Aspergillus flavus. Production of aflatoxins in sausage, salami, sucuk and kavurma by A. parasiticus and A. flavus was studied at different temperatures. A. parasiticus and A. flavus produced no aflatoxins on meat products samples at 15°C. Sucuk was a poor substrate for A. parasiticus and A. flavus at 25°C. Sausage, salami and kavurma were favorable substrates for aflatoxin production by A. parasiticus at 25°C.     

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.     

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.     

Polyphasic characterization of Aspergillus section Flavi isolated from animal feeds in Algeria

In Algeria, little information is available on the population structure of Aspergillus section Flavi in raw materials and resultant animal feeds. A total of 172 isolates belonging to Aspergillus section Flavi were recovered from 57 animal feeds and identified on the basis of macro and micro-morphological characters, mycotoxin production and genetic relatedness. For the molecular analysis, sequencing of the calmodulin gene (CaM) and the internal transcribed spacer (ITS) regions were performed for representative isolates. Four distinct morphotypes were distinguished: Aspergillus flavus (78.5%), Aspergillus tamarii (19.2%), Aspergillus parasiticus (1.7%), and Aspergillus alliaceus (0.6%). All A. flavus isolates were of the L type and no correlation between sclerotia production and aflatoxigenicity was observed. Our results showed that 68% of the A. flavus strains produced aflatoxins B (AFB), and 72.7% were cyclopiazonic acid (CPA) producers. The three isolates of A. parasiticus were able to produce AFB and aflatoxins G but not CPA whereas, all the strains of A. tamarii produced only CPA. The obtained results revealed the presence of different species of Aspergillus section Flavi, among which were aflatoxin producers. This study provides evidence useful for considerations in aflatoxin control strategies.     

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.     

Antifungal Effect of Allium tuberosum,Cinnamomum cassia,and Pogostemon cablin Essential Oils and Their Components Against Population of Aspergillus Species

Antifungal activity of Allium tuberosum (AT), Cinnamomum cassia (CC), and Pogostemon cablin (Patchouli, P) essential oils against Aspergillus flavus strains 3.2758 and 3.4408 and Aspergillus oryzae was tested at 2 water activity levels (a_w: 0.95 and 0.98). Main components of tested essential oils were: allyl trisulfide 40.05% (AT), cinnamaldehyde 87.23% (CC), and patchouli alcohol 44.52% (P). The minimal inhibitory concentration of the plant essential oils against A. flavus strains 3.2758 and 3.4408 and A. oryzae was 250 ppm (A. tuberosum and C. cassia), whereas Patchouli essential oil inhibited fungi at concentration > 1500 ppm. The essential oils exhibited suppression effect on colony growth at all concentrations (100, 175, and 250 ppm for A. tuberosum; 25, 50, and 75 for C. cassia; 100, 250, and 500 for P. cablin essential oil). Results of the study represent a solution for possible application of essential oil of C. cassia in different food systems due to its strong inhibitory effect against tested Aspergillus species. In real food system (table grapes), C. cassia essential oil exhibited stronger antifungal activity compared to cinnamaldehyde.     

Colonisation and aflatoxin formation by Aspergillus spp. on brown rices differing in endosperm properties

Aspergillus flavus Link and A. parasiticus Speare differed in sporulation and aflatoxin B₁ formation on 15 brown rices differing in starch properties and protein content. There was no significant correlation between the degree of spore formation by the fungi and the amount of aflatoxin they produced in the rice samples. Differences in sporulation and aflatoxin production were not correlated with the dimensions (weight and hardness) of brown rice nor with the protein content, alkali spreading value, amylose content and gel viscosity of milled rice. Aflatoxin B₁ was mainly concentrated in the bran layers. Histological examination revealed that the hyphae of A. flavus entered through cracks in the periderm and penetrated the endosperm.     

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.     

Selected plant essential oils and their main active components,a promising approach to inhibit aflatoxigenic fungi and aflatoxin production in food

Recent research has showed that Aspergillus flavus and Aspergillus parasiticus are aflatoxigenic species that can become very competitive in the framework of climate change. Aflatoxins show carcinogenic, mutagenic, immunotoxic and teratogenic effects on human and animals. Effective and sustainable measures to inhibit these species and aflatoxins in food are required. Origanum vulgare and Cinnamomum zeylanicum essential oils (EOs) and their major active constituents, carvacrol and cinnamaldehyde, respectively, were assayed for inhibiting these species and aflatoxin production in maize extract medium under different environmental conditions. Doses of 10–1000 mg l^?1 were assayed and the effective doses for 50 (ED₅₀) and 90% (ED₉₀) growth inhibition were determined. The ED₅₀ of cinnamaldehyde, carvacrol, oregano EO, and cinnamon EO against A. flavus were in the ranges 49–52.6, 98–145, 152–505, 295–560 mg l^?1 and against A. parasiticus in the ranges 46–55.5, 101–175, 260–425 and 490–675 mg l^?1, respectively, depending on environmental conditions. In A. flavus treatments ED₉₀ were in the ranges 89.7–90.5, 770–860 and 820–>1000 mg l^?1 for cinnamaldehyde, carvacrol and cinnamon EO, and in A. parasiticus treatments in the ranges 89–91, 855–>1000 and 900–>1000 mg l^?1, respectively. ED₉₀ values for oregano EO against both species were >1000 mg l^?1. Growth rates of both species were higher at 37 than at 25°C and at 0.99 than at 0.96 a_w. Aflatoxin production was higher at 25 than at 37°C. Stimulation of aflatoxin production was observed at low doses except for cinnamaldehyde treatments. The effectiveness of EOs and their main constituents to inhibit fungal growth and aflatoxin production in contact assays was lower than in vapour phase assays using bioactive EVOH-EO films previously reported.     

Evaluation of chemically characterised essential oils of Coleus aromaticus,Hyptis suaveolens and Ageratum conyzoides against storage fungi and aflatoxin contamination of food commodities

The result of the present investigation explores the efficacy of chemically characterised essential oils (EOs) of Coleus aromaticus, Hyptis suaveolens and Ageratum conyzoides as antifungal and antiaflatoxigenic agent against some storage fungi and the toxigenic strain of Aspergillus flavus (Saktiman 3NSt). Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of these EOs were also determined against the toxigenic strain of A. flavus (Saktiman 3NSt). The EO from C. aromaticus was found to be most effective exhibiting MIC and MFC at 0.1μL mL^?1. The EOs also completely checked aflatoxin B₁ synthesis in concentration‐dependent manner. In addition, fumigation of stored wheat samples with EOs exhibited remarkable protection (>80%) from fungal infestation showing their efficacy during in vivo storage conditions. Based on the results of the present investigation, the EOs of C. aromaticus, H. suaveolens and A. conyzoides may be recommended as novel plant‐based antifungal and aflatoxin B₁ suppressor over the synthetic preservatives.     

Effect of UV-C irradiation on inactivation of Aspergillus flavus and Aspergillus parasiticus and quality parameters of roasted coffee bean (Coffea arabica L.)

ABSTRACT

This study investigated the antifungal effect of ultraviolet-C (UV-C) against Aspergillus flavus and Aspergillus parasiticus on roasted coffee beans. Also, any changes in the quality of the roasted coffee beans were measured after UV-C irradiation. As UV-C irradiation time increased (0–2 h), the number of surviving A. flavus and A. parasiticus spores significantly (P < .05) decreased. The reduction values of A. flavus in round part, crack part, and whole roasted coffee beans were 2.16, 0.71, and 1.58 log₁₀ CFU g^?1, respectively, and the reduction values of A. parasiticus in round part, crack part, and whole roasted coffee beans were 1.03, 0.37, and 0.72 log₁₀ CFU g^?1, respectively, after 2 h of UV-C irradiation. Field emission scanning electron microscopy showed that the morphology of A. flavus and A. parasiticus spores included expanded wrinkles that were deformed by UV-C irradiation. The Hunter colours were significantly reduced (P < .05). There was no significant change (P > .05) in moisture content, but the pH was significantly decreased (P < .05). Most of the sensory parameters did not change, but there was a significant difference (P < .05) in flavour. Based on this study, 2 h of UV-C irradiation was effective in reducing 90% of A. flavus, but it was not effective against A. parasiticus present on roasted coffee beans. Also, Hunter colour, pH, and sensory parameters (flavour) were changed by UV-C irradiation.     

Assessment of Thymus vulgaris L. essential oil as a safe botanical preservative against post harvest fungal infestation of food commodities

A total of 14 odoriferous angiospermic essential oils were tested against the toxigenic strain of Aspergillus flavus. The essential oil of Thymus vulgaris L. showed highest antifungal efficacy. The thyme oil absolutely inhibited the mycelial growth of A. flavus at 0.7 μl ml^− 1 and exhibited a broad fungitoxic spectrum against eight different food contaminating fungi viz. Fusarium oxysporum, Cladosporium herbarum, Curvularia lunata, Aspergillus terreus, Aspergillus niger, Aspergillus fumigatus, Alternaria alternata and Botryodiploidia theobromae. The oil also showed significant antiaflatoxigenic efficacy as it completely arrested the aflatoxin B₁ production at 0.6 μl ml^− 1. Thyme oil as fungitoxicant was also found superior over most of the prevalent synthetic fungicides. The LC₅₀ of thyme oil against mice was recorded as 7142.85 μl kg^− 1 body weight indicating its non-mammalian toxicity and strengthening its safe exploitation as preservative for stored food commodities. The findings recommend the thyme oil as potential botanical preservative in eco-friendly control of biodeterioration of food commodities during storage.

Industrial relevance

The thyme essential oil may be recommended for large scale application as a plant based preservative for stored food items because of its strong antifungal as well as antiaflatoxigenic efficacy. Because of broad antimicrobial spectrum, more efficacy over prevalent synthetic preservatives as well as non-mammalian toxicity, the thyme essential oil may be formulated as a safe and economical plant based preservative against post harvest fungal infestation and aflatoxin contamination of food commodities.     

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.     

Antiaflatoxigenic and antioxidant activity of an essential oil from Ageratum conyzoides L.

BACKGROUND: Aflatoxin contamination of various commodities can occur as a result of infection, mainly by Aspergillus flavus and Aspergillus parasiticus. Every year, almost 25% of the world's food supply is contaminated by mycotoxins. Aflatoxins B₁, B₂, G₁ and G₂, which occur naturally, are significant contaminants of a wide variety of commodities. A number of biological activities have been associated with Ageratum conyzoides. We have therefore investigated the antiaflatoxigenic, antioxidant and antimicrobial activity of essential oils of A. conyzoides. This could help to turn A. conyzoides, a nuisance weed, into a resource. RESULTS: The essential oil of Ageratum conyzoides L. shows the presence of 12 compounds when analyzed by gas chromatography–mass spectrometry. The growth and aflatoxin production of the toxigenic strain Aspergillus parasiticus was completely inhibited by essential oil. All the studied concentrations of the oil demonstrate a reduction in mycelia growth and decreased production of different aflatoxins in fungi, as revealed by liquid chomatographic–tandem mass spectrometric analysis. Volatiles from macerated green leaf tissue of A. conyzoides were also effective against A. parasiticus. The strongest antibacterial activity was observed against the bacteria Staphylococcus aureus and Bacillus subtilis in a disk diffusion bioassay. Essential oil and methanol extract of A. conyzoides L. were assayed for their antioxidant activity. Methanol extract showed the highest antioxidant activity in FRAP and DPPH assay, whereas essential oil showed greater lipid peroxidation inhibition than methanol extract. CONCLUSION: The plant's ethno‐medicinal importance, antioxidant potential, inhibitory activity against the Aspergillus group of fungi and production of aflatoxins may add a new dimension to its usefulness in the protection of stored product. Copyright © 2010 Society of Chemical Industry     

Antifungal properties and inhibitory effects upon aflatoxin production by Zingiber officinale essential oil in Aspergillus flavus

In this study, the efficacy of ginger (Zingiber officinale Roscoe) essential oil (GEO) in reducing A. flavus growth and aflatoxin production was investigated. Gas chromatography coupled to mass spectrometry and nuclear magnetic resonance spectroscopy showed that the major components of GEO were α‐zingiberene (23.85%) and geranial (14.16%). Mycelial growth of Aspergillus flavus was reduced significantly at a GEO concentration of 150 μg mL^?1, and complete inhibition of conidial germination was observed at a concentration of 10 μg mL^?1. Statistically significant inhibition of ergosterol biosynthesis was detected at a GEO concentration of 10 μg mL^?1. GEO was capable of fully inhibiting aflatoxin production by A. flavus at a concentration of 15 μg mL^?1. The results suggest that low concentrations of GEO are capable of inhibiting aflatoxin production; such ability could be valuable in the upcoming future for agricultural companies to better control aflatoxigenic fungi in agricultural products.     

Invited review: Microbe-mediated aflatoxin decontamination of dairy products and feeds

Aspergillus flavus, Aspergillus parasiticus, and Aspergillus nomius contaminate corn, sorghum, rice, peanuts, tree nuts, figs, ginger, nutmeg, and milk. They produce aflatoxins, especially aflatoxin B₁, which is classified as a Group 1 carcinogen by the International Agency for Research on Cancer. Many studies have focused on aflatoxin removal from food or feed, especially via microbe-mediated mechanisms—either adsorption or degradation. Of the lactic acid bacteria, Lactobacillus rhamnosus GG efficiently binds aflatoxin B₁, and a peptidoglycan in the bacterium cell wall plays an important role. This ability of L. rhamnosus GG should be applied to the removal of aflatoxin B₁. Aflatoxin can be removed using other aflatoxin-degrading microorganisms, including bacterial and fungal strains. This review explores microbe-associated aflatoxin decontamination, which may be used to produce aflatoxin-free food or feed.     

Potential of pulsed electric field to control Aspergillus parasiticus,aflatoxin and mutagenicity levels: Sesame seed quality

Seed processing technologies are essential for seed safety and functionality through protection of physicochemical quality, pathogen inactivation, aflatoxin detoxification and alleviation of mutagenicity. Design of a pilot-scale unit of pulsed electric fields (PEF) to treat sesame seeds with respect to quality parameters, Aspergillus parasiticus inactivation and aflatoxin reduction as well as alleviation of aflatoxin mutagenicity were prompted in this study. PEF energy ranged from 0.97 to 17.28 J achieved maximum reductions of peroxide value and acidity number of 67.4 and 85.7%, respectively, and did not change color L*, a*, b* and hue values. A 60% reduction of A. parasiticus counts occurred at the maximum PEF energy. Aflatoxins G1, G2, B1, and B2 contents decreased by 94.7, 92.7, 86.9, and 98.7%, respectively. Except for the samples treated by 2.16 J with 100 μg/plate and by 6.80 J with 10 μg/plate, PEF treatment provided elimination of aflatoxin mutagenity. It is concluded that PEF treatment can be used to treat sesame seeds with preservation of physicochemical properties, inactivation of A. parasiticus and decomposition of aflatoxins with reduced mutagenicity.