[Inhibition of Growth of Seed-Borne Fungi and Aflatoxin Production on Stored Peanuts by Allyl Isothiocyanate Vapor].

Aspergillus parasiticus contamination of peanuts results in the production of highly toxic metabolites, such as aflatoxin B1, B2, G1 and G2, and its incidence in imported peanuts is reported to be increasing. Here, we examined whether the antifungal compound allyl isothiocyanate (AIT), which is present in mustard seed, could inhibit the growth of seed-borne fungi and aflatoxin-producing fungi. Peanuts produced in China and Japan were inoculated with A. parasiticus and exposed to AIT vapor released by a commercial mustard seed extract in closed containers under controlled conditions of temperature and humidity. AIT in the inoculated peanut samples reached its highest concentration of 44.8 ng/mL at 3 hr and decreased to 5.6 ng/mL after 9 weeks. Although AIT decreased the growth of the seed-borne fungi during the test period, the inoculated fungi survived. All tested peanuts samples were analyzed for aflatoxin using the HPLC method. There was a correlation between the number of aflatoxin-producing fungi and the total amount of aflatoxin production in the inoculated peanut samples. Our results indicate that AIT was effective in inhibiting the growth of seed-borne fungi and aflatoxin-producing fungi.

Formation Ratios of Zearalanone, Zearalenols, and Zearalanols versus Zearalenone during Incubation of Fusarium semitectum on Sorghum and Ratios in Naturally Contaminated Sorghum.

We incubated Fusarium semitectum on sorghum and measured the production of zearalenone (ZEN) and ZEN-related compounds (zearalanone (ZAN), α-zearalenol (α-ZEL), ß-zearalenol (ß-ZEL), α-zearalanol (α-ZAL) and ß-zearalanol (ß-ZAL)) in the culture by LC-MS. Of the five ZEN-related compounds, ZAN and ß-ZEL were mainly detected. The concentrations of ZEN and the five ZEN-related compounds increased until 9 days after incubation and then increased slightly or stayed constant between days 9 and 15. The ratios of α-ZEL, ß-ZEL, α-ZAL and ß-ZAL to ZEN decreased in a similar manner after 7 days, whereas the ratio of ZAN to ZEN remained constant after 5 days. Analysis of naturally contaminated sorghum by LC-MS/MS revealed that the production ratio of α-ZEL to ZEN was inconsistent with that of our in vitro incubation analysis. The results indicate that ZAN might not be suitable for use as an internal standard.

[Inhibition of aflatoxin production and fungal growth on stored corn by allyl isothiocyanate vapor].

Studies were conducted to determine the effectiveness of allyl isothiocyanate (AIT) vapor treatment with a commercial mustard seed extract (Wasaouro(®)) in controlling aflatoxin-producing fungi on stored corn. The concentration of AIT in the closed container peaked at 54.6 ng/mL on the 14th day and remained at 21.8 ng/mL on the 42nd day. AIT inhibited visible growth of aflatoxigenic molds in unsterilized corn and in sterilized corn inoculated with various aflatoxigenic fungi. However, fungi such as Aspergillus glaucus group, A. penicillioides and A. restrictus were detected by means of culture methods.

Examination of the taxonomic position of Penicillium strains used in blue cheese production based on the partial sequence of ß-tubulin.

Penicillium roqueforti is a well known starter used for blue cheese production. Two closely related species, P. carneum and P. paneum, were previously classified as varieties of P. roqueforti. Penicillium roqueforti does not produce patulin, a mycotoxin harmful for human health, whereas both P. carneum and P. paneum actively produce this toxin. From the viewpoint of food safety, it is thus important to confirm that P. carneum and P. paneum are not used for cheese production. In the present study, the taxonomic position of Penicillium strains used for blue cheese production was examined on the basis of the partial sequence of ß-tubulin. Twenty-eight Penicillium strains isolated from blue cheeses were investigated. All the examined strains belonged to P. roqueforti. Therefore, the Penicillium strains used for production of the blue cheese samples examined here do not negatively impact on human health.

Aflatoxins B and g contamination and aflatoxigenic fungi in nutmeg.

This study examined the distribution of aflatoxigenic fungi in 25 imported Indonesian nutmeg samples contaminated with aflatoxins Bs or Bs and Gs. The incidence of aflatoxigenic fungi in the samples contaminated with high levels of aflatoxin was significantly higher than that in the samples with low levels of the toxins(r=0.752). The aflatoxin production of isolates from the samples in cultures of YES broth was examined by means of TLC and HPLC analyses. The ability of isolates to produce aflatoxins did not necessarily correlate with the contamination levels of aflatoxin in the samples. We isolated aflatoxins B and G-producing fungi from 3 samples contaminated with the high levels of aflatoxins B and G. The aflatoxigenic isolates were identified as Aspergillus nomius and A. bombycis based on morphological characters, growth rates at 37°C and 42°C and also molecular-genetic methods. Our results indicate that these two species are mainly responsible for aflatoxin G contamination in nutmeg products.

Zearalenone contamination and the causative fungi in sorghum.

Natural contamination by zearalenone, a toxic metabolite of Fusarium fungi, was surveyed in 160 samples of sorghum imported from 2001 to 2006 into Japan for feed. Of these 160 samples, 84 (52.5%) were contaminated with zearalenone, ranging in concentration from 60 to 7.260 microg/kg. In the contaminated sorghum samples, F. semitectum, F. verticillioides, F. oxysporum, and other Fusarium spp. were detected. The concentration of zearalenone was well correlated with the development of colonies of F. semitectum and other Fusarium spp. When the isolates of F. semitectum and F. verticillioides were cultivated on sorghum, zearalenone was found only in F. semitectum culture. These results indicate that F. semitectum is a causal fungus of zearalenone contamination in sorghum.

[Fungal population and distribution of aflatoxigenic fungi in commercial almond powder products].

The fungal population and distribution of aflatoxin-producing fungi in 30 samples of imported almond powder products purchased from retail markets were examined in this study. Total counts of fungi ranged from under 1.0 x 10 colony-forming units (CFU)/g to 8.5 x 10(3) CFU/g as determined with the dilution plating technique. The predominant fungi in the mould-contaminated almond samples were Aspergillus niger, A. flavus and the related species, Penicillium, Cladosporium and Rhizopus. Aflatoxin-producing ability in the isolates of A. flavus and related fungi were tested by thin layer chromatography using 2% yeast extract and 15% sucrose broth culture. Four different aflatoxigenic fungi were detected in the isolates; aflatoxins B1 and B2 were produced by some strains of A. flavus and A. parvisclerotigenus, and aflatoxins B1, B2, G1, G2 were produced by all tested strains of A. parasiticus and A. nomius. Identification of the strains was based on morphological and metabolic characters.

[Mycological examination of domestic unpolished rice and mycotoxin production by isolated Penicillium islandicum].

Fungi growing on domestic rice were examined from April to June, 2003. One hundred samples of rice, which had been harvested in the autumn of 2002, were collected from the local market, and 15 samples of stored rice, which had been harvested in 2001 and stored in warehouses under government control, were used as samples. From each sample, 50 grains (100 grains in total) were plated on potato-dextrose agar (PDA) and malt yeast 40% sucrose agar (M40YA) containing chloramphenicol after being washed with sterile distilled water to remove any microorganisms on the surface, and incubated at 25 degrees C for a week. For most of the rice samples harvested in the preceding year, the proportion of grains infected with fungi was less than 20% of the total grains tested. In about half the samples of rice stored for one and half years, more than 80% of the grains were infected with fungi that grew on M40YA. The major genera of fungi isolated from the rice harvested in the preceding year were Penicillium and Alternaria, and those from the rice stored for one and a half years were Aspergillus, Penicillium and Eurotium. P. islandicum, A. versicolor, A. ochraceus and others were isolated as possible mycotoxin-producers in the mycoflora of domestic rice. P. islandicum was isolated from 3 samples, and 82% of the grains were infected with this fungus in one sample. All three isolates from these samples appeared to produce luteoskyrin on Czapek yeast extract agar, based on TLC and HPLC analysis.

Distribution of aflatoxin-producing Aspergillus flavus and Aspergillus parasiticus in sugarcane fields in the southernmost islands of Japan.

The distribution of Aspergillus flavus and Aspergillus parasiticus in sugarcane field soils and on harvested sugarcane stems was studied on seven islands of Okinawa and Kagoshima Prefectures, the southernmost prefectures in Japan. With the use of a combination of dilution plate and plant debris plate techniques, the fungi were detected on all seven islands studied and in 74% of 53 soil samples. The fungi were also found on the cut surfaces of sugarcane stems from one of the islands. A. parasiticus was the predominant fungus, although many atypical A. parasiticus isolates that produced metulated conidial heads were also obtained. The proportions of isolates testing positive for aflatoxin production were ca. 89% (146 of 164) of all isolates and ca. 69% of A. flavus isolates. More than 40% of A. flavus isolates also produced G aflatoxins. Scanning electron microscopic observation of conidial wall texture was useful in distinguishing A. parasiticus from A. flavus. Cyclopiazonic acid, an indole mycotoxin, was never synthesized by any of the A. parasiticus or G aflatoxin-producing A. flavus isolates tested.

Trends of fumonisin contamination and animal intoxication through monitoring 1991 to 1997 corn crop in the State of Paraná, Brazil.

Eleven feed samples associated with six animal (horse and poultry) intoxication outbreaks (1991) in the state of Paraná, Brazil, were evaluated for fungal and fumonisin contamination. In order to estimate the trend of livestock intoxication, fumonisin contamination was monitored in corn produced both at the commercial level (1991, 1995 crop), and in an experimental field at a local Agronomy Institute (1997 crop). The total mould count in the feed samples ranged from 2.9 x 10(3) to 1.9 x 10(7) CFU/g, with Fusarium verticillioides as the predominant species, at a high count of 2.4 x 10(4)-6.5 x 10(5) CFU/g. Fumonisins (FB1 + FB2) were detected in all corn-based feed samples at levels ranging from 2.89 to 14.54 microg/g. All 27 Northern corn samples (1991 crop) were contaminated with fumonisins at levels ranging from 2.32 to 16.64 microg/g. Twenty-six (96.3%) out of 27 corn samples from the Central-Southern region (1995 crop) were positive for fumonisins (FB1+FB2), with the range of 0.07-3.66 microg/g, while all 37 Northern samples (1995 crop) were contaminated with fumonisins ranging from 0.57 to 9.97 microg/g. Twenty-one out of 37 corn samples from the Northern region (1997 crop) were positive for fumonisins, but at low level (range of 0.05-2.67 microg/g). The results showed a decreasing trend in fumonisin contamination over the years. Nowadays animal intoxication outbreaks rarely occur in this State, as both animal producers and feed industries have become conscious about monitoring of corn and other raw materials at the quality control level.

Fungal melanin inhibitor and related compounds from Penicillium decumbens.

Two polyketides, decumbenones A and B, and versiol were isolated from the culture filtrate of the fungus, Penicillium decumbens. Their respective structures were 1-(2,8-dihydroxy-1,2,6-trimethyl-1,2,6,7,8,8a-hexahydronaphthalen-1-yl)-3-hydroxy-1-propanone and 1-(2,8-dihydroxy-1,2,6-trimethyl-1,2,4a,5,6,7,8,8a-octahydronaphthalen-1-yl)-3-hydroxy-1-propanone based on NMR spectroscopic data, chemical conversion, and X-ray analysis. Decumbenone A inhibited melanization in Magnaporthe grisea, the rice blast pathogen, whereas decumbenone B like versiol did not.