Effects of various acids and salts on growth and aflatoxin production by Aspergillus flavus NRRL 3145.

The effects of sodium chloride, sodium acetate, benzoic acid, sodium benzoate, malonic acid, and sodium malonate on growth and aflatoxin production by Aspergillus flavus were investigated in synthetic media. Sodium chloride at concentrations equivalent to or greater than 12 g/100 ml inhibited growth and aflatoxin production, while at 8 g or less/100 ml, growth and aflatoxin production were stimulated. At 2 g or less/100 ml, sodium acetate also stimulated growth and aflatoxin production, but reduction occurred with 4 g or more/100 ml. Malonic acid at 10, 20, 40, and 50 mM reduced growth and aflatoxin production (over 50%) while sodium malonate at similar concentrations but different pH values had the opposite effect. Benzoic acid (pH 3.9) and sodium benzoate (pH 5.0) at 0.4 g/100 ml completely inhibited growth and aflatoxin production. Examination of the effect of initial pH indicated that the extent of inhibitory action of malonic acid and sodium acetate was a function of initial pH. The inhibitory action of benzoic acid and sodium benzoate appeared to be a function of undissociated benzoic acid molecules. Aflatoxin reduction was usually accompanied by an unidentified orange pigment, while aflatoxin stimulation was accompanied by unidentified blue and green fluorescent spots but with lower Rf values that aflatoxins B1, G1, B2, and G2 standards.     

Influence of microbial co-inhabitants on aflatoxin synthesis of Aspergillus flavus on maize kernels

The co-inhabiting mycoflora with Aspergillus flavus observed on individual maize kernels was evaluated for its influence on aflatoxin synthesis. All 13 types of associations of different fungal species inhibited aflatoxin B₁ and G₁ production at different levels (34·3–100%). Inhibition of radial growth of A. flavus by Fusarium moniliforme (59·8%), Trichoderma viride (72·5%) and Rhizopus nigricans (42%) could be directly correlated to the per cent inhibition of aflatoxin production. High levels of inhibition of aflatoxin elaboration were noted in competition of A. flavus with other toxigenic moulds.     

Efficacy of Euphorbia splendens and Leonotis nepetaefolia on aflatoxin producing fungi Aspergillus flavus and Aspergillus parasiticus

Efficacy of three different concentrations (5, 10 and 15 mg/ml) of dry flower powder of E. splendens and L. nepetaefolia was tested on the growth of aflatoxin-producing toxigenic strains of fungi A. flavus (NCBT 101) and A. parasiticus (NCBT 128) in Sabouraud dextrose agar medium (SDA). Maximum (75%) inhibition of growth of A. flavus was seen at 15 mg/ml concentration of E. splendens flower dry powder, while A. parasiticus showed 50% inhibition of growth at 10 and 15 mg/ml concentrations. Total inhibition (100%) of growth of A. flavus was seen at 10 and 15 mg/ml for L. nepetaefolia and maximum (75%) inhibition of growth was seen for A. parasiticus at 15 mg/ml concentration. Bioassay with groundnut seeds soaked with different concentrations of flower extract proved that both fungi were incapable of infecting the seeds at 10 and 15 mg/ml of L. nepetaefolia flower extracts.     

The effect of clove and cinnamon oils on growth of and aflatoxin production by Aspergillus flavus

The effect of different concentrations of clove and cinnamon oils was studied on the growth of and aflatoxin production by Aspergillus flavus in SMKY liquid medium. The effect of these compounds was also verified against aflatoxin production in maize. Significant reduction (P < 0.05) in the elaboration of aflatoxin in liquid culture after treatment with more than 100 μg ml^-1 of these compounds was recorded. Cinnamon oil exhibited maximum inhibitory action and reduced 78% aflatoxin formation on maize at 1000 mg kg^-1.     

Inhibition of growth and aflatoxin biosynthesis of Aspergillus flavus by extracts of some Egyptian plants

The effect of five different concentrations (2, 4, 6, 8 and 10 mg ml-1) of an aqueous extracts of Lupinus albus, Ammi visnaga and Xanthium pungens were tested on growth and aflatoxin production by Aspergillus flavus in a chemically defined medium. All the plants inhibited mycelial growth and aflatoxin formation. The inhibitory effect was proportional with the applied concentration. Growth and aflatoxin production appeared to be correlated processes. The nature of the plant extract also affected the ratio of B1 to B2, and there was no correlation between the inhibition of aflatoxins or growth of the fungus and the resultant B1: B2 ratio.     

Isolation, purification, and antibiotic activity of o-methoxycinnamaldehyde from cinnamon.

o-Methoxycinnamaldehyde has been isolated and purified from powdered cinnamon. The compound inhibits the growth and toxin production of mycotoxin-producing fungi. The substance completely inhibited the growth of Aspergillus parasiticus and A. flavus at 100 microgram/ml and A. ochraceus and A. versicolor at 200 microgram/ml. It inhibited the production of aflatoxin B1 by over 90% at 6.25 microgram/ml, ochratoxin A at 25 microgram/ml, and sterigmatocystin at 50 microgram/ml. The substance also displayed a strong inhibitory effect on the growth of five dermatophytoses species, e.g., Microsporum canis (minimum inhibitory concentration, 3.12 to 6.25 microgram/ml). However, no antibacterial effect was observed at concentrations as high as 50 microgram/ml.     

Regional differences in production of aflatoxin B1 and cyclopiazonic acid by soil isolates of aspergillus flavus along a transect within the United States

Soil isolates of Aspergillus flavus from a transect extending from eastern New Mexico through Georgia to eastern Virginia were examined for production of aflatoxin B1 and cyclopiazonic acid in a liquid medium. Peanut fields from major peanut-growing regions (western Texas; central Texas; Georgia and Alabama; and Virginia and North Carolina) were sampled, and fields with other crops were sampled in regions where peanuts are not commonly grown. The A. flavus isolates were identified as members of either the L strain (n = 774), which produces sclerotia that are >400 micrometer in diameter, or the S strain (n = 309), which produces numerous small sclerotia that are <400 micrometer in diameter. The S-strain isolates generally produced high levels of aflatoxin B1, whereas the L-strain isolates were more variable in aflatoxin production; variation in cyclopiazonic acid production also was greater in the L strain than in the S strain. There was a positive correlation between aflatoxin B1 production and cyclopiazonic acid production in both strains, although 12% of the L-strain isolates produced only cyclopiazonic acid. Significant differences in production of aflatoxin B1 and cyclopiazonic acid by the L-strain isolates were detected among regions. In the western half of Texas and the peanut-growing region of Georgia and Alabama, 62 to 94% of the isolates produced >10 microgram of aflatoxin B1 per ml. The percentages of isolates producing >10 microgram of aflatoxin B1 per ml ranged from 0 to 52% in the remaining regions of the transect; other isolates were often nonaflatoxigenic. A total of 53 of the 126 L-strain isolates that did not produce aflatoxin B1 or cyclopiazonic acid were placed in 17 vegetative compatibility groups. Several of these groups contained isolates from widely separated regions of the transect.     

The effect of eucalyptus oil on growth and aflatoxin production by Aspergillus flavus

The effect of eucalyptus oil on growth and aflatoxin production by Aspergillus flavus was tested at three levels, viz . 0·05, 0·1 and 0·2 ml/50 ml SMKY medium. After 6 days of incubation on 0·05 and 0·1 ml supplemented SMKY medium, growth and toxin production were inhibited while at 0·2 ml concentration there was no growth. However, after 12 days of incubation toxin production was greater than the controls.     

The influences of fungitoxicants on growth and aflatoxin production by Aspergillus flavus

The influence of six fungitoxicants on growth and aflatoxin production by Aspergillus flavus was tested in liquid SMKY medium at two concentrations, viz . 0.1 and 0.5%. Thiram completely inhibited the aflatoxin production at 0.5% concentration. Other fungitoxicants showing more than 60% inhibition were bavistin and daconil. Vitavax (0.1%) and agrosan GN (0.1 and 0.5%) stimulated the growth of fungus and aflatoxin elaboration after 7 d of incubation. Dithane M-45 moderately inhibited aflatoxin synthesis. Treatment with fungitoxicants also alters the ratio of B₁ and G₁.     

Fungicidal and anti-aflatoxigenic effects of the essential oil of Cymbopogon citratus (DC.) Stapf. (lemongrass) against Aspergillus flavus Link. isolated from stored rice

AIMS: To develop a natural fungicide against aflatoxigenic fungi, to protect stored rice, using the essential oil of lemongrass. METHODS AND RESULTS: Aspergillus flavus Link. was isolated from stored rice and identified as an aflatoxigenic strain. Lemongrass oil was tested against A. flavus and the test oil was fungistatic and fungicidal against the test pathogen at 0.6 and 1.0 mg ml(-1), respectively. Aflatoxin production was completely inhibited at 0.1 mg ml(-1). The results obtained from the thin layer chromatographic bioassay and gas chromatography indicated citral a and b as the fungicidal constituents in lemongrass oil. During the fumigant toxicity assay of lemongrass oil, the sporulation and the mycelial growth of the test pathogen were inhibited at the concentrations of 2.80 and 3.46 mg ml(-1), respectively. CONCLUSION: Lemongrass oil could be used to manage aflatoxin formation and fungal growth of A. flavus in stored rice. SIGNIFICANCE AND IMPACT OF THE STUDY: Currently, fungicides are not used to control fungal pests or mycotoxin production on stored rice. Rice treated with the essential oil of lemongrass could be used to manage fungal pests as well as the insect pests in stored rice. The essential oil is chemically safe and acceptable to consumers, as synthetic chemical fungicides can cause adverse health effects to consumers.     

Antimicrobial activity and inhibition of aflatoxin B1 formation by olive plant tissue constituents

Ethanolic extracts of olive callus tissues, added at 0.5 or 1.0% to media on which Aspergillus flavus was grown, inhibited aflatoxin production by 90% without inhibiting the fungal growth. The extract was found to contain mainly caffeic acid and, to a lesser extent, catechin and coumarins. The fungicidal and bactericidal activity of caffeic acid, catechin, coumarin and p-, o- or m-coumaric acid were tested and only caffeic acid and o-coumaric acid inhibited aflatoxin production. The inhibitory effect had no correlation with the growth of the fungus. Only coumarin at 10 mmol/1 totally inhibited fungal growth. Of the phenolic constituents of callus tissues tested, catechin and caffeic acid (10 mmol/l) showed bactericidal activity towards Pseudomonas aeruginosa and Staphylococcus aureus.     

Antimicrobial activity and inhibition of aflatoxin B1 formation by olive plant tissue constituents

Ethanolic extracts of olive callus tissues, added at 0.5 or 1.0% to media on which Aspergillus flavus was grown, inhibited aflatoxin production by 90% without inhibiting the fungal growth. The extract was found to contain mainly caffeic acid and, to a lesser extent, catechin and coumarins. The fungicidal and bactericidal activity of caffeic acid, catechin, coumarin and p-, o- or m-coumaric acid were tested and only caffeic acid and o-coumaric acid inhibited aflatoxin production. The inhibitory effect had no correlation with the growth of the fungus. Only coumarin at 10 mmol/1 totally inhibited fungal growth. Of the phenolic constituents of callus tissues tested, catechin and caffeic acid (10 mmol/1) showed bactericidal activity towards Pseudomonas aeruginosa and Staphylococcus aureus.     

Effect of pyridazinone herbicides on growth and aflatoxin release by Aspergillus flavus and Aspergillus parasiticus

The influence of pyridazinone herbicides on aflatoxin production by Aspergillus flavus and A. parasiticus was studied in liquid media. Mycelia production was not affected by 20, 40, or 60 micrograms of herbicide per ml; however, aflatoxin production by A. parasiticus was higher in media with herbicide, whereas A. flavus produced lower aflatoxin levels.     

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.     

Effect of pyridazinone herbicides on growth and aflatoxin release by Aspergillus flavus and Aspergillus parasiticus.

The influence of pyridazinone herbicides on aflatoxin production by Aspergillus flavus and A. parasiticus was studied in liquid media. Mycelia production was not affected by 20, 40, or 60 micrograms of herbicide per ml; however, aflatoxin production by A. parasiticus was higher in media with herbicide, whereas A. flavus produced lower aflatoxin levels.     

The influence of colouring and pungent agents of red Chilli (Capsicum annum) on growth and aflatoxin production by Aspergillus flavus

Capsanthin and capsaicin, the colouring and pungent principles of red chilli Capsicum annum , respectively, were tested against the growth and aflatoxin producing potentials of Aspergillus flavus in SMKY liquid medium. Capsanthin completely checked both the growth and toxin production at all the concentrations viz. 0.2, 0.6 and 1.0 mg ml^-1, till the fourth day of incubation. On the 10th day growth of the fungus and toxin biosynthesis were 39 and 22% of the control, respectively, at 1.0 mg ml^-1. Capsaicin showed some inhibitory efficacy only up to the fourth day of incubation. The fungus grew thereafter with a marginal inhibition in growth at the highest concentration. The amount of the toxin in the medium was also higher.     

Inhibition in aflatoxin biosynthesis by the extract of Amorphophallus campanulatus (OL) and calcium oxalate

A complete inhibition in aflatoxin production by Aspergillus flavus was observed with greater than 4.5 mg ml^-1 of 20 g 100 ml^-1 leaf extract of Amorphophallus campanulatus (OL). Suppression in growth was also well pronounced (81% at 8.0 mg ml^-1). The comparative efficacy of the corm of the plant was lower. Calcium oxalate, an important constituent of 'OL', completely checked the growth and toxin biosynthesis at 0.4 mg ml^-1 concentration.     

Mold flora and aflatoxin contamination of stored and cooked samples of pearl millet in the Paharia tribal belt of Santhal paragana, Bihar, India

Stored and cooked samples of pearl millet (Pennesetum typhoides), which is regularly consumed as food by the Paharia tribe in the hilly regions of Santhal Pargana, Bihar State, India, that were harvested in January 1989 were analyzed for mold flora, natural occurrence of Aspergillus flavus and A. parasiticus, and incidence and levels of aflatoxin B1. Of the 22 fungal species isolated, A. flavus and A. parasiticus were the predominant species (63.8%) during the rainy season, followed by other species of Aspergillus, Penicillium, Fusarium, Rhizopus, Helminthosporium, and Curvularia. Screening of 169 A. flavus and A. parasiticus strains showed that 59 of them were toxigenic, producing various combinations of aflatoxins B1, B2, G1, and G2. The amounts of aflatoxin B1 ranged between 4 and 30 mg/100 ml of liquid medium. Analysis of stored and cooked samples also revealed a high incidence and alarming levels of naturally produced aflatoxin B1. Forty-nine of 75 stored and 16 of 38 cooked samples contained various combinations of aflatoxins. The levels of aflatoxin B1 ranged between 17 and 2,110 ppb in stored samples and 18 and 549 ppb in cooked samples. The correlation of insect damage with A. flavus and A. parasiticus incidence and quantity of aflatoxin B1 was found to be insignificant.     

Mold flora and aflatoxin contamination of stored and cooked samples of pearl millet in the Paharia tribal belt of Santhal paragana, Bihar, India.

Stored and cooked samples of pearl millet (Pennesetum typhoides), which is regularly consumed as food by the Paharia tribe in the hilly regions of Santhal Pargana, Bihar State, India, that were harvested in January 1989 were analyzed for mold flora, natural occurrence of Aspergillus flavus and A. parasiticus, and incidence and levels of aflatoxin B1. Of the 22 fungal species isolated, A. flavus and A. parasiticus were the predominant species (63.8%) during the rainy season, followed by other species of Aspergillus, Penicillium, Fusarium, Rhizopus, Helminthosporium, and Curvularia. Screening of 169 A. flavus and A. parasiticus strains showed that 59 of them were toxigenic, producing various combinations of aflatoxins B1, B2, G1, and G2. The amounts of aflatoxin B1 ranged between 4 and 30 mg/100 ml of liquid medium. Analysis of stored and cooked samples also revealed a high incidence and alarming levels of naturally produced aflatoxin B1. Forty-nine of 75 stored and 16 of 38 cooked samples contained various combinations of aflatoxins. The levels of aflatoxin B1 ranged between 17 and 2,110 ppb in stored samples and 18 and 549 ppb in cooked samples. The correlation of insect damage with A. flavus and A. parasiticus incidence and quantity of aflatoxin B1 was found to be insignificant.     

Aflatoxin B1 and fumosin B1 in mixed cultures of Aspergillus flavus and Fusarium proliferatum on maize

Production of aflatoxin B1 and fumonisin B1 in pure and mixed cultures of Aspergillus flavus and Fusarium proliferatum were determined on irradiated maize seeds inoculated with different spore concentrations at 0.97 water activity (a(w)) and a temperature of 25 degrees C. The highest levels of aflatoxin B1 were produced by A. flavus at the lowest levels of inoculum (10(3) spore ml(-1)). There was no spore concentration influence on fumonisin B1 production after 10, 20 and 35 days of incubation. When A. flavus was co-inoculated with F. proliferatum, aflatoxin B1 production was inhibited. The higher the inocula levels of Fusarium produced, the higher the inhibition and this inhibition increased during the incubation period. Total inhibition was reached at 35 days of incubation. There was no interaction influence on fumonisin B1 production at all inoculum levels assayed. These results suggest that under optimal environmental conditions of substrate, water activity and temperature, the interaction between A. flavus and F proliferatum could produce inhibition of aflatoxin B1 and stimulation of fumonisin B1.