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Correlation of growth and aflatoxin production by Aspergillus flavus with some essential metals in gamma irradiated crushed corn.

The effects of gamma irradiation and some essential metals on growth and aflatoxin B1 production by Aspergillus flavus in crushed corn were investigated. The production of aflatoxin by A. flavus was influenced by the addition of zinc, copper or iron and the effect gradually decreased with increasing metal concentration from 0 to 300 ppm. A. flavus grew and depleted zinc, copper and iron at initial concentration of 100, 200 or 300 ppm. Presence of 100 ppm zinc, copper or iron plus gamma irradiation (0.5, 1.0, 2.0 kGy) enhanced the growth of A. flavus and the production of aflatoxin in contrast with irradiated samples alone. A. flavus was able to metabolize and deplete elements in all gamma-irradiated samples. These results suggest that stricter control of element levels in gamma irradiated grains could control aflatoxin contamination.

Aflatoxin B1↗

Mycoflora of hazelnut (Corylus avellana L.) and aflatoxin content in hazelnut kernels artificially infected with Aspergillus parasiticus.

In this research, 30 hazelnut samples were used to determine their internal mould populations and their aflatoxin contents. The results showed that Aspergillus was the most common genus in the experimental samples (96.6% of all samples). Thirty-one% of isolates were classified as Aspergillus flavus and none of the samples contained aflatoxins. In the second part of the experiment, the kernels were inoculated with conidia of Aspergillus parasiticus and incubated at three different humidified conditions and two different temperatures for 45 days. Aflatoxin contents of the samples kept in 98% relative humidity and at 28 degrees C were higher (904.6 micrograms/kg) than that of the other samples. On the other hand, no aflatoxin was detected in the control samples that were not inoculated but kept at the same conditions.

Aflatoxins↗

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.

Aflatoxin B1↗

The fate of aflatoxins during the production of "Ogiri", a West African fermented melon seed condiment from artificially contaminated seeds.

Twenty-six market samples and four laboratory-prepared samples of "ogiri" were screened for aflatoxin contamination. Aflatoxins were not detected in any of the samples. The fermented product (ogiri) was prepared with Aspergillus flavus-contaminated melon seeds. Losses of 64.7% aflatoxin B1 and 82.9% aflatoxin G1 were observed at the end of the third day of fermentation of the ground melon seeds. The samples were completely detoxified at the fourth day of fermentation. Increase in pH of the mash from 6.2 to 7.2 was observed during fermentation.

Aflatoxins↗

Effect of gamma-irradiation on aflatoxin B1 production by Aspergillus flavus and chemical composition of three crop seeds.

The effect of gamma-irradiation on aflatoxin B1 production by Aspergillus flavus, and the chemical composition of some different crop seeds were investigated. A. flavus infected seeds behaved differently according to their principal constituents. A. flavus caused an increase in protein and decrease in lipids and carbohydrate contents of wheat, soyabean and fababean seeds. Growth of A. flavus and production of aflatoxin B1 was inhibited at a dose level of 5 kGy. A. flavus utilizes carbohydrates of seeds for its growth and aflatoxin production. Crops were arranged, in descending order, according to aflatoxin produced in seeds as wheat > soyabean > fababean. There were no changes in chemical constituents of irradiated seeds, such as protein, lipids, and carbohydrates.

Aflatoxin B1↗

Elevated aflatoxin exposure and increased risk of hepatocellular carcinoma.

To elucidate the importance of aflatoxin in the etiology of hepatocellular carcinoma (HCC), a community-based cohort study combined with molecular dosimetry of aflatoxin exposure was performed in the Penghu Islets where the HCC mortality rate is highest in Taiwan. A total of 6,487 residents aged 30 to 65 years were recruited in the two-stage screening survey and underwent regular follow-up examination. Among 33 newly diagnosed HCC cases, 31 (94%) were chronic hepatitis B surface antigen (HBsAg) carriers and 3 (9%) were positive for antibodies against hepatitis C virus (HCV). Among 20 HCC patients and 86 matched healthy controls whose serum samples were tested for aflatoxin B1 (AFB1)-albumin adducts by competitive enzyme-linked immunosorbant assay (ELISA), 13 (65%) HCC patients and 32 (37%) matched controls were seropositive, showing a statistically significant multivariate-adjusted odds ratio of 5.5 with a 95% confidence interval of 1.2 to 24.5. The results imply the elevated risk of HCC among Penghu residents may be attributable to their heavy exposure to aflatoxins and high HBsAg carrier rate.

Adult↗

Dietary aflatoxins and human liver cancer. A study in Swaziland.

A study in Swaziland to assess the possible relationship of aflatoxin contamination and the incidence of primary liver cancer is reported. Aflatoxin ingestion levels have been determined in "food from the plate" samples collected over a 1-year period. A significant correlation between the calculated ingested daily dose and the adult male incidence of primary liver cancer in different parts of Swaziland has been established. Samples of foodstuffs other than the plate samples also reflected the correlation of aflatoxin contamination and liver cancer. This study extends and amplifies the findings of an earlier study in the Murang'a district of Kenya and supports the hypothesis that aflatoxin ingestion is a factor in the genesis of primary liver cancer in Africa.

Aflatoxins↗

Fungal contamination, natural occurrence of mycotoxins and resistance for aflatoxin accumulation of some broad bean (Vicia faba L.) cultivars.

Using glucose-Czapek's and 10% NaCl glucose-Czapek's agar media incubated at 28 +/- 2 degrees C, nine genera and 18 species were identified from 10 different cultivars seed of broad bean. Aspergillus and Penicillium were the most common genera on the two types of media used. The most common fungal species were Aspergillus flavus, A. niger, A. tamarii and Penicillium chrysogenum. Thin-layer chromatographic analysis indicated that seeds of one cultivar only was naturally contaminated with aflatoxins B1, B2, G1 and G2. Nine mycotoxin-free cultivars of broad bean were examined for resistance or susceptibility for aflatoxin production when inoculated by Aspergillus flavus IMI 89717. Results obtained revealed that seeds of two cultivars were highly resistant, two cultivars showed partial resistance and the remaining were susceptible to the establishment of A. flavus and aflatoxin accumulation. The results did not show any relationship between morphological characters (colour, shape and size) and the amount of aflatoxin produced on the different broad bean seed cultivars. Also the results reveal absence of significant variation in the total nitrogen of the highly susceptible, partially resistant and highly resistant groups of broad bean seeds examined. Mean values of calcium, total phosphate and potassium of both seed coat and seed kernel of the susceptible seeds were low as compared to those of partially and highly resistant seed cultivars. The susceptible seeds contained higher levels of magnesium, zinc and sodium while the resistant seeds contained lower ones.

Aflatoxins↗

Action of carbamate biocides on sterols, gibberellin and aflatoxin formation.

An interrelationship between primary and secondary metabolites of a toxigenic mould (Aspergillus parasiticus var. globosus IMI 120920) was obtained in liquid broth containing various concentrations of dithane Z-78 and propoxur. The results show that a stimulatory response was recorded in gibberellin, protein, sterol and aflatoxin yield in cultures treated with 15, 30, 50 and 100 ppm a.i. of dithane Z-78. On the other hand, propoxur reduced the yield of gibberellin, amino acids, protein, sterol and aflatoxin. In comparison to its effect on the mycelial growth, gibberellin and aflatoxin activity is inhibited more severely than dry weight increase, thus leading to the assumption that there is a specific effect of the inhibitor on both gibberellin and aflatoxin synthesis.

Aflatoxins↗

Development and validation of a method for the quantitative determination of aflatoxin contaminants in Maytenus ilicifolia by HPLC with fluorescence detection.

A method for the quantification of aflatoxins B1, G1, B2 and G2 in the medicinal herb Maytenus ilicifolia was developed and validated. The method used immunoaffinity columns for sample clean-up and HPLC with fluorescence detection without any derivatisation step. The method showed good inter-day accuracy (bias values in the range 4.5-10.7%) and precision (5-16% RSD) when applied to the determination of levels of aflatoxins ranging from 7 to 20 ppb in the plant material. The detection limits for samples of the plant material spiked with aflatoxins were 3.5 ng/g for B1 and G1 and 0.1 ng/g for B2 and G2. The method was successfully applied to commercial samples of Maytenus ilicifolia for the screening of aflatoxin contaminants.

Aflatoxins↗

Control of Aspergillus growth and aflatoxin production using natural maize phytochemicals under different conditions of water activity.

The effects of the natural phytochemicals trans-cinnamic acid (CA) and ferulic acid (FA) alone at concentrations of 1-25 mM and in 16 combinations (M: mixtures) on growth and aflatoxin B(1) production by Aspergillus flavus Link and A. parasiticus Speare were evaluated. Studies on growth rate and aflatoxin B(1) production were carried out in vitro in relation to a water activity a(w) of 0.999, 0.971, 0.955 and 0.937. Overall, CA at concentrations of 10 and 20 mM and FA-CA mixtures M3 (20 + 5 mM respectively), M8 (25 + 5 mM), M9 (1 + 10 mM), M10 (10 + 10 mM), M11 (20 + 10 mM), M12 (25 + 10 mM), M13 (1 + 20 mM), M14 (10 + 20 mM), M15 (20 + 20 mM) and M16 (25 + 20 mM) were the treatments most effective at inhibiting growth of the four species assayed. All strains were much more sensitive to all natural phytochemicals tested on growth rate at a(w) = 0.937. CA and the FA-CA mixtures M1 (1 + 1 mM respectively), M4 (25 + 1 mM), M5 (1 + 5 mM), M6 (10 + 1 mM), M7 (20 + 1 mM), M8 (25 + 5 mM), M9 (1 + 10 mM), M10 (10 + 10 mM), M11 (20 + 10 mM), M12 (25 + 10 mM), M13 (1 + 20 mM), M14 (10 + 20 mM), M15 (20 + 20 mM) and M16 (25 + 20 mM) completely inhibited aflatoxin B(1) production by all strains at a(w) = 0.999, 0.971, 0.955 and 0.937. Decreased aflatoxin B(1) levels in comparison with the control were observed with FA at 1, 10, 20 and 25 mM with the strains RCM89, RCM108 and RCM38 at a(w) = 0.971, 0.955 and 0.999 respectively. The data show that CA and FA can be considered as effective fungitoxicants for A. flavus and A. parasiticus in in vitro assay. The information obtained is part of an ongoing study to determine their application at the storage level.

Aflatoxins↗

Picroliv protects against aflatoxin B1 acute hepatotoxicity in rats.

The hepatoprotective activity of Picroliv, a standardized iridoid glycoside fraction of Picrorhiza kurroa, has been investigated by studying the protection of biochemical and histological changes induced in livers of rats given single oral doses (7 mg/kg) of aflatoxin B1. Administration of aflatoxin B1 resulted in a significant increase in 5'-nucleotidase, r-glutamyl transpeptidase, acid ribonuclease, total lipids, cholesterol and lipid peroxides in liver and transaminases, alkaline phosphatase and bilirubin in serum. However, the activity of glucose 6-phosphatase and levels of cytochrome P450, cytochrome b5, DNA, RNA, proteins and glycogen in liver and total proteins in serum decreased. The liver histology showed ballooned hepatocytes, degeneration, microvesicular fat, focal necrosis, bile duct hyperplasia and proliferation of oval and spindle cells in portal tracts. When Picroliv (25 mg/kg x 7 days) was given to aflatoxin B1 toxicated rats, the majority of the biochemical and histological changes were significantly protected. The findings indicate a hepatoprotective activity of Picroliv against aflatoxin B1 toxicity in rats.

Aflatoxin B1↗

Radioactivity in urine and feces of mink (Mustela vison) treated with [14C] aflatoxin B1.

Excretion of radioactivity by mink (Mustela vison) during 7 days after intraperitoneal injection of two different amounts of aflatoxin B1 was studied. Male mink that received a single dose of 25 mug aflatoxin B1/kg body weight excreted an average of 89.5% of administered radioactivity (56.8% via feces, 32.7% via urine); whereas female mink excreted an average of 85% (63.6% via feces, 21.4% via urine) of administered radioactivity during the 7-day period. Male and female mink given 150 mug aflatoxin B1/kg body weight excreted an average of 76.9-80.1% of administered radioactivity during the 7 days that followed treatment with toxin. These mink excreted somewhat more of the administered radioactivity in their urine than did the mink that received the lower dose of aflatoxin (37.2 vs. 32.7% for males and 32.7 vs. 21.4% for females). Regardless of sex and dosage of toxin, most of the radioactivity ultimately excreted either through feces or urine appeared in the first 24 h after toxin was administered to mink.

Aflatoxins↗

Production of aflatoxins on soybean and cottonseed meals.

p6e production of aflatoxins by Aspergillus flavus SRRC-1000 growing on soybean (Forrest) and glandless cottonseed (Deltapine 16) meals was examined with respect to effects of zinc and phytate. Aflatoxins were not produced on unautoclaved soybean meal. Addition of zinc (as zinc sulfate) to autoclaved meal inhibited aflatoxin production and supplementation with sodium phytate relieved this inhibition. Addition of sodium phytate alone promoted production. When cottonseed meal was treated to release native phytate into the meal from phytate-sequestering globoids, aflatoxin production increased. However, the largest production on cottonseed meal occurred upon dialysis of the meal without releasing phytate, implying removal of a small molecular weight inhibitor.

Aflatoxins↗

Effect of selected inhibitors on growth, pigmentation, and aflatoxin production by Aspergillus parasiticus.

We have treated a wild type strain of Aspergillus parasiticus with several known aflatoxin inhibitors in hopes of finding specific metabolic blocks in the aflatoxin biosynthetic pathway. In defined medium, benzoic acid (2 and 3 mg/ml), cinnamon (1 mg/ml), and sodium acetate (5 mg/ml) were fungitoxic. Benzoic acid (0.5 and 1 mg/ml), chlorox (5 microliters/ml), and dimethyl sulfoxide (5 microliters/ml) did not affect dry weight or mycelial pigmentation. Sodium benzoate (1, 2, 4 and 8 mg/ml) added after 2 days growth inhibited aflatoxin production in two defined media. We were unable to confirm previously published reports that an uncharacterized yellow pigment accumulates with benzoate-inhibition of aflatoxin biosynthesis.

Acetates↗

Different media and methodologies for the detection of aflatoxin production by Aspergillus flavus strains isolated from trout feed.

We have studied the aflatoxin producing capacity of 41 Aspergillus flavus strains isolated from the mycoflora present of natural media (wheat, rice and mixed feed) synthetic medium (Aflatoxin Producing Ability Medium) and semisynthetic media (Coconut Agar Medium and Glucose Yeast Extract Agar) were compared. Aflatoxins were analysed on days 4 and 8 post-inoculation under an incubation temperature of 28 degrees C. A total of 30 strains (75.7%) were producers on natural media as detected by Thin Layer Chromatography: 23 strains on wheat, 27 on rice and 12 on mixed feed. The results by qualitative fluorescence tests on synthetic and semisynthetic media were: 3 strains positive on Coconut Agar Medium (CAM) 1 on Glucose Yeast Extract Agar (GY + Agar) and none on Aflatoxin Producing Ability Medium (APA).

Aflatoxins↗

Experimental short time production of aflatoxins by Aspergillus parasiticus in mixed feeds as related to various moisture contents.

Experimental short time production of aflatoxins in mixed feeds at 22, 28 and 37 degrees C as related to various moisture contents was studied. Growth of Aspergillus parasiticus was not observed in the meals with a moisture content ranging around 15% (22, 28 and 37 degrees C); the lowest quantifiable total aflatoxins at the fourth day was detected at 22 degrees C with 19.4% of moisture content; the higher total quantity of aflatoxins (113 mg/kg) was produced at 28 degrees C with 29.3% of moisture content. The ratio aflatoxin B1/aflatoxin G1 increased as the temperature raised.

Aflatoxins↗

Survey of aflatoxin-producing fungi in certain fermented foods and beverages in Thailand.

Aflatoxin-producing fungi were found in fermented foods and beverages: fermented rice (kaomak), soybean sauce (taotjo), peanut butter, soy sauce (shoyu), Thai red and white wine, and rice sugar wine. These foods were extracted directly and tested for aflatoxins by thin-layer chromatography (TLC) and high pressure liquid chromatography (HPLC). Four strains of aflatoxin-producing fungi were isolated from peanut butter, taotjo, and shoyu. Direct extracts of 10% of the peanut butters tested and 5% of the kaomak tested contained large amounts of aflatoxins. The HPLC procedures used in this experiment utilized chloroform-ethyl acetate (3:1).

Aflatoxins↗