Modelling the effect of temperature and water activity on the growth rate of Aspergillus flavus and aflatoxin production in peanut meal extract agar.

Aspergillus flavus is the predominant species that produce aflatoxins in stored peanuts under favourable conditions. This study aimed to describe the growth and aflatoxin production by two A. flavus strains isolated from imported raw peanuts and to model the effects of temperature and aw on their colony growth rate as a function of temperature and aw in Peanut Meal Extract Agar (PMEA). A full factorial design with seven aw levels (0.85-0.98 aw) and five temperature levels (20-40 °C) was used to investigate the growth and aflatoxin production. Colony diameter was measured daily for 28 days while AFB1 and total aflatoxin were determined on day 3, 7, 14, and 21. The maximum colony growth rate, µmax (mm/day) was estimated by using the primary model of Baranyi, and the µmax was then fitted to the secondary model; second-order polynomial and linear Arrhenius-Davey to describe the colony growth rate as a function of temperature and aw. The results indicated that both strains failed to grow at temperature of 20 °C with aw <0.94 and aw of 0.85 for all temperatures except 30 °C. The highest growth rate was observed at 30 °C, with 0.98 aw for both strains. The analysis of variance showed a significant effect of strain, temperature, and aw on the fungal growth and aflatoxin production (p < 0.05). Furthermore, both secondary models were in good agreement with the observed µmax. However, the polynomial model was found to be a better predictor of the experimental data. A similar pattern was observed in aflatoxin production but in a narrower range of temperature (25-35 °C) and aw (0.92-0.98 aw). The highest production of aflatoxins was observed on day 21 at 30 °C with the aw level of 0.98 for both strains. Overall, the current findings may help in improving the mycotoxin management and intervention strategies in peanuts, especially during storage.

Antioxidant-related catalase CTA1 regulates development, aflatoxin biosynthesis, and virulence in pathogenic fungus Aspergillus flavus.

Reactive oxygen species (ROS) induce the synthesis of a myriad of secondary metabolites, including aflatoxins. It raises significant concern as it is a potent environmental contaminant. In Aspergillus flavus., antioxidant enzymes link ROS stress response with coordinated gene regulation of aflatoxin biosynthesis. In this study, we characterized the function of a core component of the antioxidant enzyme catalase (CTA1) of A. flavus. Firstly, we verified the presence of cta1 corresponding protein (CTA1) by Western blot analysis and mass-spectrometry based analysis. Then, the functional study revealed that the growth, sporulation and sclerotia formation significantly increased, while aflatoxins production and virulence were decreased in the cta1 deletion mutant as compared with the WT and complementary strains. Furthermore, the absence of the cta1 gene resulted in a significant rise in the intracellular ROS level, which in turn added to the oxidative stress level of cells. A further quantitative proteomics investigation hinted that in vivo, CTA1 might maintain the ROS level to facilitate the aflatoxin synthesis. All in all, the pleiotropic phenotype of A. flavus CTA1 deletion mutant revealed that the antioxidant system plays a crucial role in fungal development, aflatoxins biosynthesis and virulence.

Antifungal and anti-aflatoxigenic activity of Heliopsis longipes roots and affinin/spilanthol against Aspergillus parasiticus by downregulating the expression of alfD and aflR genes of the aflatoxins biosynthetic pathway.

In the present study, ethanolic extract from Heliopsis longipes roots and affinin/spilanthol against Aspergillus parasiticus growth and aflatoxins production were studied in relation to the expression of aflD and aflR, two key genes of aflatoxins biosynthetic pathway. Phytochemical analysis of the ethanolic extract by GC-EIMS identified affinin/spilanthol (7.84 ± 0.27 mg g-1) as the most abundant compounds in H. longipes roots. The antifungal and anti-aflatoxigenic assays showed that affinin/spilanthol at 300 µg mL-1 produced the higher inhibition of radial growth (95%), as well as, the higher aflatoxins production inhibition (61%) in comparison to H. longipes roots (87% and 48%, respectively). qRT-PCR revealed that the expression of aflD and aflR genes showed a higher downregulation in affinin/spilanthol at 300 µg mL-1. The expression ratio of alfD was suppressed by affinin/spilanthol in 79% and aflR in 84%, while, a lower expression ratio suppressed by H. longipes was obtained, alfD (55%) and aflR (59%). Affinin/spilanthol possesses higher antifungal and anti-aflatoxigenic activity against A. parasiticus rather than H. longipes roots, and this anti-aflaxotigenic activity occurring via downregulation of the aflD and aflR genes. Thus, H. longipes roots and affinin/spilanthol can be considered potent antifungal agents against aflatoxigenic fungus, especially, affinin/spilanthol.

Preparation, characterization and anti-aflatoxigenic activity of chitosan packaging films incorporated with turmeric essential oil.

Here, we studied the preparation, characterization, anti-aflatoxigenic activity, and molecular mechanism in vitro of chitosan packaging films containing turmeric essential oil (TEO). First, we took the mechanical properties as the evaluation Index, screened for the optimum preparation conditions of packaging films with 1.5 µL/cm2 TEO using single factor and orthogonal experiments, and characterized the film properties. We found that the addition of TEO affected the microcosmic structure of films and advanced water resistance capacity. In addition, we investigated the inhibitory effects of pure chitosan films and packaging films containing 1.5 µL/cm2 or 3.0 µL/cm2 TEO on the growth and conidial formation of Aspergillus flavus (A. flavus, CGMCC 3.4410), as well as the accumulation of aflatoxin over the course of seven days. We found that the packaging films possessed a prominent antifungal activity on A. flavus. Finally, we discuss preliminary results surrounding gene expression of packaging films which inhibit aflatoxin biosynthesis. The expressions levels of 16 genes related to aflatoxin biosynthesis were found to be either completely or almost completely inhibited. Therefore, the addition of the natural antifungal agent TEO in chitosan packaging films represent a remarkable method to significantly promote the development and application of antifungal packaging materials.

Activation of Aflatoxin Biosynthesis Alleviates Total ROS in Aspergillus parasiticus.

An aspect of mycotoxin biosynthesis that remains unclear is its relationship with the cellular management of reactive oxygen species (ROS). Here we conduct a comparative study of the total ROS production in the wild-type strain (SU-1) of the plant pathogen and aflatoxin producer, Aspergillus parasiticus, and its mutant strain, AFS10, in which the aflatoxin biosynthesis pathway is blocked by disruption of its pathway regulator, aflR. We show that SU-1 demonstrates a significantly faster decrease in total ROS than AFS10 between 24 h to 48 h, a time window within which aflatoxin synthesis is activated and reaches peak levels in SU-1. The impact of aflatoxin synthesis in alleviation of ROS correlated well with the transcriptional activation of five superoxide dismutases (SOD), a group of enzymes that protect cells from elevated levels of a class of ROS, the superoxide radicals (O2-). Finally, we show that aflatoxin supplementation to AFS10 growth medium results in a significant reduction of total ROS only in 24 h cultures, without resulting in significant changes in SOD gene expression. Our findings show that the activation of aflatoxin biosynthesis in A. parasiticus alleviates ROS generation, which in turn, can be both aflR dependent and aflatoxin dependent.

4-Hydroxy-7-methyl-3-phenylcoumarin Suppresses Aflatoxin Biosynthesis via Downregulation of aflK Expressing Versicolorin B Synthase in Aspergillus flavus.

Naturally occurring coumarins possess antibacterial and antifungal properties. In this study, these natural and synthetic coumarins were used to evaluate their antifungal activities against Aspergillus flavus, which produces aflatoxins. In addition to control antifungal activities, antiaflatoxigenic properties were also determined using a high-performance liquid chromatography in conjunction with fluorescence detection. In this study, 38 compounds tested and 4-hydroxy-7-methyl-3-phenyl coumarin showed potent antifungal and antiaflatoxigenic activities against A. flavus. Inhibitory mode of antiaflatoxigenic action by 4-hydroxy-7-methyl-3-phenyl coumarin was based on the downregulation of aflD, aflK, aflQ, and aflR in aflatoxin biosynthesis. In the cases of coumarins, antifungal and aflatoxigenic activities are highly related to the lack of diene moieties in the structures. In structurally related compounds, 2,3-dihydrobenzofuran exhibited antifungal and antiaflatoxigenic activities against A. flavus. The inhibitory mode of antiaflatoxigenic action by 2,3-dihydrobenzofuran was based on the inhibition of the transcription factor (aflS) in the aflatoxin biosynthesis pathway. These potent inhibitions of 2,3-dihydrobenzofuran and 4-hydroxy-7-methyl-3-phenyl coumarin on the Aspergillus growth and production of aflatoxins contribute to the development of new controlling agents to mitigate aflatoxin contamination.

Aflatoxin biosynthesis control produced by Aspergillus flavus in layer hens feed during storage period of six months.

Aflatoxins (AFTs) are a group of closely related toxins that are produced by different fungus species. Food and feed contamination with AFT is a worldwide health-related problem. As a result of fungal attack, the food and feed resulted in a principal socioeconomic loss and toxins produced in feed and food items harm the humans and animals in different ways. The anti-aflatoxigenic effect Psidium guajava, Ficus benghalensis, Gardenia radicans, Punica granatum and Ziziphus jujuba leaves were evaluated against aflatoxins (AFTs), produced by Aspergillus flavus in layer feed during storage. Among the investigated medicinal plant leaves, P. granatum showed highly promising anti-aflatoxigenic activity and completely inhibited the AFTs (B1 and B2) production over storage period without compromising the nutritive quality of feed (ash, protein, fat, fiber, Fe, Ca, P and K contents). Leaves of F. benghalensis and Z. jujuba were also effective however, higher concentration (15%) inhibited the AFTs production up to 99% and also maintained nutritive quality of feed. G. radicans was found least effective in controlling the AFTs production. Results revealed that all plant leaves were effective in controlling AFTs production in layer feed over the storage period of six months and these plants are potential candidate to replace the fungicides used to protect feed and other agricultural commodities from AFTs production during storage.

Mechanisms of antifungal and anti-aflatoxigenic properties of essential oil derived from turmeric (Curcuma longa L.) on Aspergillus flavus.

The antifungal activity and potential mechanisms in vitro as well as anti-aflatoxigenic efficiency in vivo of natural essential oil (EO) derived from turmeric (Curcuma longa L.) against Aspergillus flavus was intensively investigated. Based on the previous chemical characterization of turmeric EO by gas chromatography-mass spectrometry, the substantially antifungal activities of turmeric EO on the mycelial growth, spore germination and aflatoxin production were observed in a dose-dependent manner. Furthermore, these antifungal effects were related to the disruption of fungal cell endomembrane system including the plasma membrane and mitochondria, specifically i.e. the inhibition of ergosterol synthesis, mitochondrial ATPase, malate dehydrogenase, and succinate dehydrogenase activities. Moreover, the down-regulation profiles of turmeric EO on the relative expression of mycotoxin genes in aflatoxin biosynthetic pathway revealed its anti-aflatoxigenic mechanism. Finally, the suppression effect of fungal contamination in maize indicated that turmeric EO has potential as an eco-friendly antifungal agent.

Isolation, selection and evaluation of antagonistic yeasts and lactic acid bacteria against ochratoxigenic fungus Aspergillus westerdijkiae on coffee beans.

UNLABELLED: In this study, yeasts and lactic acid bacteria (LAB) were isolated from coffee fruits and identified via biochemical and molecular approaches. The isolates represented the Pichia, Debaryomyces, Candida, Clavispora, Yarrowia, Sporobolomyces, Klyveromyces, Torulaspora and Lactobacillus genera. Four isolates, namely Pichia fermentans LPBYB13, Sporobolomyces roseus LPBY7E, Candida sp. LPBY11B and Lactobacillus brevis LPBB03, were found to have the greatest antagonist activity against an ochratoxigenic strain of Aspergillus westerdijkiae on agar tests and were selected for further characterization. Applications of P. fermentans LPBYB13 in coffee cherries artificially contaminated with A. westerdijkiae showed efficacy in reducing ochratoxin A (OTA) content up to 88%. These results highlight that P. fermentans LPBYB13 fulfils the principle requirements of an efficient biological control of aflatoxigenic fungi in coffee beans and may be seen as a reliable candidate for further validation in field conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Studies based on microbial ecology and antagonistic interactions are important for the development of new strategies in controlling aflatoxin contamination of crops and are relevant to further biotechnological applications. This study shows that coffee fruit is a potential source for the isolation of microbial strains with antifungal ability. A new yeast strain, Pichia fermentans LPBYB13, showed efficacy in reducing growth and ochratoxin A production of Aspergillus westerdijkiae in coffee beans. Our results should encourage the use of this yeast strain on a large scale for biocontrol of aflatoxigenic fungi in coffee beans.

Utilization of waste fruit-peels to inhibit aflatoxins synthesis by Aspergillus flavus: a biotreatment of rice for safer storage.

Antifungal activity in lemon and pomegranate peels was considerable against Aspergillus flavus, higher in pomegranate (DIZ 37mm; MIC 135µg/mL). Powdered peels (5, 10, 20% w/w) were mixed in inoculated rice. The inhibitory effect on fungal-growth and production of aflatoxins by A. flavus was investigated at storage conditions - temperature (25, 30°C) and moisture (18%, 21%) for 9months. The maximum total aflatoxins accumulated at 30°C, 21% moisture and at 25°C, 18% moisture were 265.09 and 163.45ng/g, respectively in control. Addition of pomegranate-peels inhibited aflatoxins production to 100% during four month-storage of rice at 25°C and 18% moisture, while lemon-peels showed similar inhibitory effect for 3months at same conditions. However a linear correlation was observed in aflatoxins level with temperature and moisture. Studies showed that both fruit-wastes are potent preventer of aflatoxin production in rice, useful for a safer and longer storage of rice.

Evaluation of allium and its seasoning on toxigenic, nutritional, and sensorial profiles of groundnut oil.

Mitigation of xerophilic storage fungi-associated aflatoxin threat in culinary oil will be a new technology advantage to food industries. Groundnut oil isolate Aspergillus flavus MTCC 10680 susceptibility to Allium species (A. sativum L., A. cepa L., and A. cepa var. aggregatum) extracts, composition, and in silico confirmation of extract's phytoconstituent aflatoxin synthesis inhibition were determined. The behavior of seasoning carrier medium groundnut oil in the presence of Allium was also determined. All the Allium species extracts exhibited concentration dependent in vitro inhibition on mycelial biomass, radial growth, and toxin elaboration. The gas chromatography-mass spectrometry revealed the presence of 28, 16, and 9 compounds in the extracts of A. sativum, A. cepa, A. cepa var. aggregatum, respectively. The Allium phytocostituents-like hexadecanoic acid, 5-Octanoyl-2,4,6(1H,3H,5H)-pyrimidinetrione, Guanosine, and so on, showed higher binding energy with aflatoxin synthesis key enzyme ver1. Allium seasoning increased the typical nutty odor of the groundnut oil with sweet aroma note as well as intensification of pale yellow color. Allium seasoning exhibited the highest aflatoxin detoxification and aroma development without any nutritional loss. Culinary oil Allium seasoning has anti-aflatoxin and food additive potential for use in food industries.

Effects of Heracleum persicum ethyl acetate extract on the growth, hyphal ultrastructure and aflatoxin biosynthesis in Aspergillus parasiticus.

The ethyl acetate extract of leaves, seeds and flowers of Heracleum persicum, a medicinal plant of Iran (family Apiaceae) inhibited growth and aflatoxin (AF) production of Aspergillus parasiticus. On the basis of total dry weight growth inhibition by the leaf extract ranged from 17.1 to 36.9 %, by the flower extract from 32.2 to 75.6 %, and by the seed extract from 27.5 to 74.9 %. Production of AFB1 and AFG1 was inhibited in a dose-dependent manner, with a reduction of 88.5-100 % at the highest concentration of 8,000 µg/ml tested. The flower extract decreased ergosterol content of hyphae most significantly. Electron microscopy further revealed structural defects in the treated A. parasiticus including disruption of cytoplasmic membranous compartments, detachment of plasma membrane from the cell wall, and disorganization of hyphal compartments. Collapsed hyphae without conidiation, shorter branches and undifferentiated hyphal tips were also evident. The results indicate that H. persicum extract exerts antifungal and anti-AF activities by disrupting plasma membrane integrity and permeability mainly through interference with ergosterol biosynthesis. These results show that H. persicum can serve as a potent and safe alternative for inhibiting toxigenic aspergilli growth and thus preventing AF contamination of foods and feeds.

Inhibitory effect of the essential oil of Curcuma longa L. and curcumin on aflatoxin production by Aspergillus flavus Link.

Aflatoxins are highly toxic, mutagenic, teratogenic and carcinogenic mycotoxins. Consumption of aflatoxin-contaminated food and commodities poses serious hazards to the health of humans and animals. Turmeric, Curcuma longa L., is a native plant of Southeast Asia and has antimicrobial, antioxidant and antifungal properties. This paper reports the antiaflatoxigenic activities of the essential oil of C. longa and curcumin. The medium tests were prepared with the oil of C. longa, and the curcumin standard at concentrations varied from 0.01% to 5.0%. All doses of the essential oil of the plant and the curcumin standard interfered with mycotoxin production. Both the essential oil and curcumin significantly inhibited the production of aflatoxins; the 0.5% level had a greater than 96% inhibitory effect. The levels of aflatoxin B(1) (AFB(1)) production were 1.0 and 42.7 µg/mL, respectively, for the samples treated with the essential oil of C. longa L. and curcumin at a concentration of 0.5%.

Inhibitory effects of gossypol, gossypolone, and apogossypolone on a collection of economically important filamentous fungi.

Racemic gossypol and its related derivatives gossypolone and apogossypolone demonstrated significant growth inhibition against a diverse collection of filamentous fungi that included Aspergillus flavus, Aspergillus parasiticus, Aspergillus alliaceus, Aspergillus fumigatus, Fusarium graminearum, Fusarium moniliforme, Penicillium chrysogenum, Penicillium corylophilum, and Stachybotrys atra. The compounds were tested in a Czapek agar medium at a concentration of 100 µg/mL. Racemic gossypol and apogossypolone inhibited growth by up to 95%, whereas gossypolone effected 100% growth inhibition in all fungal isolates tested except A. flavus. Growth inhibition was variable during the observed time period for all tested fungi capable of growth in these treatment conditions. Gossypolone demonstrated significant aflatoxin biosynthesis inhibition in A. flavus AF13 (B(1), 76% inhibition). Apogossypolone was the most potent aflatoxin inhibitor, showing greater than 90% inhibition against A. flavus and greater than 65% inhibition against A. parasiticus (B(1), 67%; G(1), 68%). Gossypol was an ineffectual inhibitor of aflatoxin biosynthesis in both A. flavus and A. parasiticus. Both gossypol and apogossypolone demonstrated significant inhibition of ochratoxin A production (47%; 91%, respectively) in cultures of A. alliaceus.

Effect of Equisetum arvense and Stevia rebaudiana extracts on growth and mycotoxin production by Aspergillus flavus and Fusarium verticillioides in maize seeds as affected by water activity.

Cereals are a very important part of the human and animal diets. However, agricultural products can be contaminated by moulds and their mycotoxins. Plant extracts, particularly those of Equisetum arvense and Stevia rebaudiana have been reported previously to contain antioxidant compounds which may have antifungal properties. In this study, E. arvense and S. rebaudiana extracts were tested for their control of mycotoxigenic fungi in maize. The extracts were tested separately and as a mixture for their effect on growth of Aspergillus flavus and Fusarium verticillioides. Extracts were added to unsterilised inoculated maize at different water activity (a(w)) levels (0.85-0.95). Moulds were inoculated and incubated for 30 days. Results confirmed that the extract of E. arvense and a mixture 1:1 of Equisetum-Stevia may be effective for the inhibition of both growth of A. flavus and aflatoxin production at high water activity levels (pre-harvest conditions). In general, growth of the F. verticillioides was reduced by the use of plant extracts, especially at 0.95 a(w). However, fumonisin presence was not significantly affected. E. arvense and S. rebaudiana extracts could be developed as an alternative treatment to control aflatoxigenic mycobiota in moist maize.

Inhibition of aflatoxin production and growth of Aspergillus parasiticus by Cuminum cyminum, Ziziphora clinopodioides, and Nigella sativa essential oils.

Aflatoxins are highly toxic and carcinogenic metabolites produced by Aspergillus parasiticus on food and agricultural commodities. Natural products may control the production of aflatoxins. The aims of this study were to evaluate the effects of the essential oils (EOs) of Cuminum cyminum, Ziziphora clinopodioides, and Nigella sativa on growth and aflatoxins production by A. parasiticus. Minimal inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs) of the EOs were determined and compared with each other. Determination of aflatoxins (AFB(1), AFB(2), AFG(1), and AFG(2)) was performed by immunoaffinity column extraction using reverse phase-high performance liquid chromatography. The major oil components were α-pinene (30%) in C. cyminum, pulegone (37%) in Z. clinopodioides, and trans-anthol (38.9%) in N. sativa oils. In broth microdilution method, C. cyminum oil exhibited the strongest activity (MIC(90): 1.6; MFC: 3.5 mg/mL), followed by Z. clinopodioides (MIC(90): 2.1; MFC: 5.5 mg/mL) and N. sativa (MIC(90): 2.75; MFC: 6.25 mg/mL) oils against A. parasiticus (p<0.05). Aflatoxin production was inhibited at 0.25 mg/mL of C. cyminum and Z. clinopodioides oils, of which that of C. cyminum was a stronger inhibitor. C. cyminum EO caused significant reductions in values of 94.2% for AFB(1), 100% for AFB(2), 98.9% for AFG(1), 100% for AFG(2), and 97.5% for total aflatoxin. It is concluded that the EOs of C. cyminum, Z. clinopodioides, and N. sativa could be used as natural inhibitors in foods at low concentrations to protect from fungal and toxin contaminations by A. parasiticus.

Antimicrobial activity of Gynostemma pentaphyllum extracts against fungi producing aflatoxin and fumonisin and bacteria causing diarrheal disease.

Gynostemma pentaphyllum was investigated to determine its antimicrobial activities against human.and animal pathogens that produce aflatoxin, fumonisin, and diarrheal disease. The fungi were Aspergillusflavus, Aspergillus parasiticus and Fusarium verticillioides. The bacteria were Vibrio, Salmonella, Shigella, Escherichia coli and Staphylococcus aureus. G. pentaphyllum was extracted by five different methods. The obtained extracts were designated Extracts A, B, C, D and E. The results of the antifungal assay against A.flavus andA. parasiticus showed Extracts A and B at 10,000 ppm inhibited growth at 8-28%. Extracts A and B at 10,000 ppm also showed activity against F. verticillioides at 41-43%. Extract A, B and C were able to inhibit the tested strains better than the Extracts D and E. The MIC values of the extracts against gram-negative bacteria ranged from

Effect of Capsicum carotenoids on growth and aflatoxins production by Aspergillus flavus isolated from paprika and chilli.

The aim of this study was to determine the effect of a carotenoid mixture (Capsantal FS-30-NT), containing capsanthin and capsorubin, on growth and aflatoxins (AF) production of AF-producing Aspergillus flavus isolates. Each isolate, previously isolated from paprika and chilli, was inoculated on Czapek Yeast extract Agar (CYA) medium supplemented with different amounts of capsantal (0-1%) and incubated at 10, 15 and 25 °C during 21 days. Growth rates and lag phases were obtained, and AF production was determined at 7, 14 and 21 days. None of the isolates grew at 10 °C and one isolate (UdLTA 3.193) hardly grew at 15 °C. Capsantal addition had no effect over lag phases and growth rates at 15 °C. At 25 °C capsantal reduced growth rates and increased lag phases. However, the effect of capsantal on AF production was inconclusive, because it depended on temperature or time, and most of the times it was not significant. Low temperature has been a crucial factor in AF production, regardless of the capsantal concentration tested. Industrial storage temperature for paprika and chilli use to be approximately 10 °C, so if this temperature is maintained mould growth and AF production should be prevented.

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 B(1), B(2), G(1) and G(2), 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.

Survey of Thymus migricus essential oil on aflatoxin inhibition in Aspergillus flavus.

Essential oil components as result of non host disease resistance of plants have high capability to introduce as alternative of chemical pesticides. Thymus migricus essential oil was selected to investigation of its antifungal activity on survival and growth of Aspergillus flavus. For obtain essential oil first Leaves and flowers of Th. migricus collected then dried. The Essential oil was extracted by means of hydro-distillation and afterwards GC-MS analysis was performed to identify their components. The main constituents that resulted were Thymol (44.9%), Geraniol (10.8%), gamma-Terpinene (10.3%), Citronellol (8.5%) and p-Cymene (7.2%). EC50 and MIC (Minimum Inhibitory Concentration) of Th. migricus oil against A. flavus was 324.42 microl/l and 451.62 microl/l, respectively. Whereas EC50 and MIC for chemical thiabendazol was 650 microl/l and 1635 microl/l, respectively. The EC50 and MIC concentrations of Th. migricus oil in antifungal activity examination were used in aflatoxin inhibition test. Result of HPTLC measurement showed that both of concentrations inhibit aflatoxin production completely compares to control with 7.63 ppm aflatoxin production. In other word, Th. migricus oil can suppress aflatoxin production in concentrations lower than EC50 for mycelium growth.