Deterioration and spoilage of peanuts and desiccated coconuts from two sub-Saharan tropical East African countries due to the associated mycobiota and their degradative enzymes

A broad variety of fungi (84 species belonging to 36 genera) were identified with more taxa infesting peanut seed samples from two tropical countries (29 genera and 61 species) compared to those found in desiccated coconuts (20 genera and55 species) on both DRBC and DG18 media. This may be due to the higher moisture levels in peanuts (5.07–7.97%) compared with coconuts (1.5–4.17%). More taxa and propagules were recovered on DG18 in both cases. The dominant fungi from both substrates on both isolation media were Aspergillus andPenicillium, with other fungi from only one substrate/medium. The aflatoxigenic species (A. flavus) dominated Kenyan samples more so than Ugandan samples on both substrates. However only71.5% and 87% of the peanut kernels, on DRBC and DG18, respectively, were found to be infested with fungi. The aflatoxigenic species (A. flavus/parasiticus)were found in 75% of the samples, however only 15.75% and 13%of the kernels analyzed were infested. The most frequently isolated species from peanuts were A. niger followed by A. flavus and M. phaseolina. E. repens, E. amstelodami,E. rubrum and E. chevalieri dominated peanut seeds on DG18, and R. stolonifer, A. parasiticus,F. solani, L. theobromae and P. chrysogenumon DRBC. The mean count of fungal propagules in coconut samples were approximately 0.7× 10³ and 0.8× 10³on DRBC and DG18, respectively, with a high proportion of those propagules recorded for the aflatoxigenic species (about 0.17×10³ and 0.25× 10³ colonies/g). The mycobiota of desiccated coconut was dominated by A. niger, A. flavusand P. chrysogenum. Also A. ochraceu,P. waksmanii, Paecilomyces variotii,P. islandicum and R. mucilaginosa were more frequent on DRBC, while, species of Cladosporium,Chrysosporium and Eurotium were more frequent on DG18. Enzyme indices (or the activities) for each specific strain, when determined after 5 and 8 days of incubation, proved to be similar. A recommendation is given. The proteolytic and lipolytic potentialities of the most commonly encountered species from both peanuts and coconuts were studied. The most interesting observation is that most of the positive isolates, in both commodities had high enzymic activity compared to those reported earlier for isolates of the same species. Such capabilities suggest that these commodities are expected to deteriorate, since climatic conditions in tropical areas favour fungal proliferation. Emphasis on the proper harvesting, drying, handling, transportation and/or storage; and also education of the populace, especially those are dealing with these foods, should be taken into consideration by the relevant authorities. The contaminated foods constitute a health hazard for human consumption. This revised version was published online in June 2006 with corrections to the Cover Date.     

Studies on mycoflora of salt marshes in Egypt. IV=osmophilic fungi

Ninety-five species and seven varieties which belong to thirty-nine genera were isolated from 74 soil samples from salt marshes. At 28°C, on 30% sucrose Czapek's agar, ninety species and four varieties which belong to thirty-six genera were encountered, from which A. niger, A. fumigatus A. terreus and P. notatum were the most frequent. On 60% sucrose Czapek's agar, sixty-three species and three varieties were recovered which belong to twenty-five genera, from which A. niger, A. fumigatus, Cladosporium herbarum and A. terreus were the most frequent.At 45°C, on 30% sucrose, sixteen species and four varieties were identified, but on 60% sucrose, fourteen species and three varieties were isolated. A. fumigatus and A. niger were the most frequent on both sucrose concentrations.The results reveal that the soil samples poor in total osmophilic fungi (at 28°C) were significantly higher in their content of total soluble salts, Na and K and significantly lower in the average number of species per soil samples than the rich ones; the difference was nonsignificant in case of organic matter.     

Survey of the mycoflora and mycotoxins of cotton seeds and cotton seed products in Egypt

Thirty-nine species and 16 fungal genera were isolated from Egyptian cotton seeds, cotton seed meal and cotton seed cake on 1% glucose-Czapek's agar medium incubated at 28 °C. Aspergillus was the most frequent genus and it emerged in 87–100% of the samples contributing 70–98% of total fungi in the three substrates tested. The most common species were A. niger, A. flavus, A. fumigatus, A. terreus and Rhizopus stolonifer; A. niger, A. fumigatus and Penicillium corylophilum; and A. niger, A. flavus, A. terreus, A. nidulans and Rhizopus stolonifer, respectively. Cotton seeds and cotton seed products were naturally contaminated by aflatoxin B₁ and B₂. About 16% of the different substrates tested were positive for aflatoxin contamination. No citrinin, ochratoxin A, patulin, sterigmatocystin, diacetoxyscirpenol, T-2 toxin or zearalenone were detected in the samples assayed.     

Microbial flora of some soils of Mawson Base and the Vestfold Hills,Antarctica

Summary A microbiological survey of 40 soils from contaminated and pristine localities of Mawson Base or the Vestfold Hills (Davis Base) is described. High microbial populations (from 10⁶ to in excess of 10⁹ cells g^-1 dry soil) were associated with soil contaminated by man or animals, soil from beneath a moss bed, or soil from beneath translucent quartz pebbles and containing green algae. Lower microbial populations (generally between 10⁴–10⁶ cells g^-1 soil) were associated with pristine soils lacking an identifiable primary producer. No correlation was evident between population density and soil particle size, moisture content, pH or salinity as determined by the electrical conductivity (E.C.) of soil suspensions, although relationships were evident between the components of the microflora and E.C. Microbial groups associated with low E.C. were chitinolytic bacteria, fungi and green algae; favoured by high E.C. were halotolerant bacteria. The microflora of uncontaminated soils appeared to be dominated by Gram-negative aerobic rods (68% of the total flora) with Moraxella, Acinetobacter and Pseudomonas being the most frequently isolated genera. The remaining microflora comprised Grampositive pleomorphic bacteria (mainly coryneforms), 13%; filamentous Gram-negative aerobic rods (morphologically akin to Bacillus species), 12%; and yeasts, 7%. Streptomyces was identified among the dominant microflora of only one soil. Nitrogen-fixing bacteria were never isolated, although their presence was indicated in 40% of the soils tested by acetylene-reduction assay. Anaerobes were not detected in any soil using dilutionplate techniques, but were occasionally detected by enrichment in semi-solid media. All of 242 isolates obtained at 10° incubation were able to grow at 15°, 3.3% failed to grow at 18° and a further 9.5% failed to grow at 25°. Of 304 isolates obtained on one-tenth strength Trypticase-Soy agar (0.05% nutrient concentration), 10% failed to grow on the full-strength medium. Soils from the vicinity of Mawson or Davis bases contained a variety of fungi (Cladosporium, Chrysosporium, Acremonium, Phoma, Aureobasidium and Trichoderma species being isolated) in contrast to pristine soils, from which only Verticillium was occasionally isolated.Abbreviations E.C. electrical conductivity - TSA Trypticase-Soy agar - m.S. milli Siemens     

Assessment of mycoflora and infestation of insects,vector of <Emphasis Type="Italic">Aspergillus</Emphasis> section <Emphasis Type="Italic">Flavi</Emphasis>, in stored peanut from Argentina

The occurrence of spoilage fungi and Aspergillus section Flavi populations, the aflatoxins incidence, the role of insects as vectors of mycotoxin-producing fungi and the AFs-producing ability of the isolated species throughout the peanut (Arachis hypogaea L.) storage period were evaluated. Analyses of fungal populations from 95 peanut seed samples did not demonstrate significant differences between the incidences in each sampling period. Aspergillus section Flavi were isolated during all incubation periods. Cryptolestes spp. (Coleoptera: Cucujidae) were collected in August, September and October with 18, 16 and 28% of peanut samples contaminated, respectively. Insects isolated during August showed 69% of Aspergillus section Flavi contamination. A. flavus was the most frequently isolated (79%) from peanut seeds and from insect (59%). The greater levels of AFB₁ were detected in September and October with a mean of 68.86 μg/kg and 69.12 μg/kg respectively. The highest proportion of A. flavus toxigenic strains (87.5%) was obtained in June. The presence of Aspergillus section Flavi and insect vectors of aflatoxigenic fungi presented a potential risk for aflatoxin production during the peanut storage period. Integrated management of fungi and insect vectors is in progress.     

Occurrence of Aspergillus section Flavi and aflatoxin B1 in corn genotypes and corn meal in Argentina

A study has been carried out in Argentina on samples of corn genotypes from a breeding station as well as in commercially available corn meal. All samples were analyzed for fungal infection and aflatoxin B₁.Mycological analysis of corn genotypes showed the presence of three principal genera of filamentous fungi Fusarium (100%), Penicillium (67%) and Aspergillus (60%). In the genus Fusarium three species were identified, F. moniliforme (42%), F. nygamai (56%) andF. proliferatum (1.8%). Eight species ofPenicillium were identified, the predominant species isolated were P. minioluteum, P. funiculosum and P. variabile. In the genus ranked third in isolation frequency, two species were identified, A. flavus and A. parasiticus, the percentage of infection was 78% and 21%, respectively. Only one corn genotype was contaminated with aflatoxin B₁ at a level of 5 ppb. The cornmeal samples showed great differences in fungal contamination, the values ranging from 1 × 10¹ to 7 × 10⁵ cfu g^–1. Fusarium (68%), Aspergillus (35%) and Penicillium (21%) were the most frequent genera isolated. Among the genus, Aspergillus, A. parasiticus (38%) was the most frequent species isolated. All the samples of corn meal were negative to aflatoxin B₁. These results indicate a low degree of human exposure to aflatoxins in Argentina through the ingestion of maize or corn meal.This revised version was published online in October 2005 with corrections to the Cover Date.     

Implications of fungal infections and mycotoxins in camel diseases in Saudi Arabia

Natural feed ingredients (corn, barley and wheat bran) and compound feed (manufactured pellet) are two types of fodder used for animal feeding, especially camel in Saudi Arabia. Twenty samples of each type of fodder were collected from seven different regions and screened for the presence of fungi, aflatoxins, ochratoxin and zearalenone. Fungal isolation of natural feed ingredients yielded 10 genera and 38 species of different fungi. Compound fodder samples were contaminated with 16 genera and 32 species of fungi. Total counts of Aspergillus, Penicillium and Fusarium in the animal feed samples were ranged from 54 to 223 × 10³, 31.9 to 60 × 10³ and 18 to 29 × 10³ CFU/g, respectively. These isolates when tested for aflatoxin, ochratoxin and zearalenone producing ability, revealed this property in only four isolate, identified as Aspergillus flavus, A. parasiticus, A. ochraceus and Fusarium graminaerum. The percentage of toxigenic fungi was ranged from 5.5% to 30% for natural feed ingredients and from 4.5% to 20% for compound feed. The incidence of aflatoxins (AFT) in samples of natural feed ingredients was found to be ranged from 1 to 24.8 ppb, ochratoxin A (OTA) ranged from 1 to 44 ppb and zearalenone (ZON) ranged from 1 to 23 ppb. Contamination of compound feed with aflatoxin and ochratoxin A was ranged from 1 to 6.4 ppb and 1 to 4.7 ppb, respectively. All samples collected were found contaminated with fungi or their toxins and natural feed samples were more contaminated compared to compound feed samples. The concentrations detected were in the allowed limit (<20 ppb) except four samples of natural feed ingredients which were above the allowed limit of the tested mycotoxins. In conclusion, feed samples were contaminated with fungi and some toxigenic isolates which were responsible about mycotoxin production. Some samples had exceeded amount of AFT, OTA and ZON and may be contaminated with other mycotoxins which mean implication of fungi in camel health problems and death in Saudi Arabia.     

Fungal contamination of some imported spices

Seventeen imported raw spice samples obtained from retail outlets were examined for spoilage mould profile. A total of 665 fungal isolates, representing 14 species, were recovered and identified from dried and ground spice samples on several media using standard dilution plate method. Moisture content varied greatly among various samples and were generally high. The most heavily contaminated spice samples examined were observed in red chili and black pepper in order of magnitude of 1580 and 1120 cfu/g, respectively. The most predominant fungal genera encountered were Aspergillus, Penicillium, Rhizopus, Cladosporium and Trichoderma. Yeasts were also frequently recovered, but not identified. Relative occurrence values of taxa disclosed ranged between 36.4% for A. flavus and 0.6% for A. parasiticus and Absidia corymbifera. Samples obtained from gunny bags encounter higher colony counts and contamination frequency than other packing methods. The present magnitude of contamination and spectra of mycobiota approximate those reported for similar spice samples. Although several potentially mycotoxigenic fungi were isolated during the present study, neither the foodstuff nor the fungi were assayed for the presence of these toxins.     

Screening of white-rot fungi for their ability to mineralize polycyclic aromatic hydrocarbons in soil

Soil samples from an agricultural field contaminated with 10 ppm¹⁴C-benz(a)anthracene in glass tubes were brought into contact with cultures of wood-rotting fungi, precultivated on wheat straw substrate. Forty-five strains of white-rot fungi and four brown-rot fungi were tested for their ability to colonize the soil and to mineralize¹⁴C-benz(a)anthracene to¹⁴CO₂ within a 20-week incubation time. Twenty-two white-rot fungi and all brown-rot fungi were unable to colonize the soil. Twenty-three strains of white-rot fungi, all belonging to the genusPleurotus, colonized the soil. During the experiment the noncolonizing fungi and their substrate disintegrated more and more to a nonstructured pulp from which water diffused into the soil. The same phenomenon was observed in the control which contained only straw without fungus and contaminated soil. In samples with colonizing fungi the substrate as well as the mycelia in the soil remained visibly unchanged during the entire experiment. Surprisingly, most samples with fungi not colonizing the soil and the control without fungus liberated between 40 and 58 % of the applied radioactivity as¹⁴CO₂ whereas the samples with the colonizing fungi respired only 15–25 % as¹⁴CO₂. This was 3–5 times more¹⁴CO₂ than that liberated from the control (4.9 %) which contained only contaminated soil without straw and fungus. A similar result was obtained with selected colonizing and noncolonizing fungi and soil contaminated with 10 ppm¹⁴C-pyrene. However, in pure culture studies in which¹⁴C-pyrene was added to the straw substrate,Pleurotus sp. (P2), as a representative of the colonizing fungi, mineralized 40.3 % of the added radioactivity to¹⁴CO₂. The noncolonizing fungiDichomitus squalens andFlammulina velutipes liberated only 17.2 or 1.7 %, respectively, as¹⁴CO₂. These results lead to the hypothesis that the native soil microflora stimulated by the formed products of straw lysis is responsible for high degradation rates found with noncolonizing fungi.     

Fungal flora and mycotoxins of six kinds of nut seeds for human consumption in Saudi Arabia

A wide range of moulds representing several genera and species, was recorded in this study from 5 seed samples of each almond, cashew nut, chestnut, hazelnut, pistachio nut and walnut collected from different markets in Ar' Ar, Saudi Arabia. The total counts of fungi were widely fluctuated between 1960–7704 and 1948–7434 colonies/g dry seeds on glucose-Czapek's and glycerol agar media at 28°C, respectively, and represented twenty genera, 53 species and 2 varieties of fungi. The prevalent fungi on the 2 agar media wereAspergillus flavus, A. niger andPenicillium chrysogenum. On glucose-Czapek's agar,Rhizopus stolonifer andAspergillus flavus var.columnaris were isolated from all 6 kinds of nut,A. parasiticus from 5 kinds andA. fumigatus from 4 kinds with high frequencies.Eurotium species were completely absent on glucose-Czapek's agar but they were isolated in high frequency from all kinds of nut on glycerol agar medium. The different nut samples were analyzed by thin layer chromatography for the presence of aflatoxins B₁, B₂, G₁ & G₂, citrinin, ochratoxins, patulin, sterigmatocystin, diacetoxyscirpenol, T-2 toxin and zearalenone. Aflatoxins B₁ & G₁ were detected in 3 out of the 5 samples tested of chestnut at concentrations ranging between 20 to 60 µg/kg. All other samples of almond, cashew nut, hazelnut, pistachio nut, and walnut that were analyzed were mycotoxin free.     

Occurrence of Ochratoxin A-Producing Fungi in Commercial Corn Kernels in Argentina

Potentially ochratoxigenic Aspergillus and Penicillium species were identified and the natural occurrence of ochratoxin A (OTA) in corn kernels was evaluated. Likewise, the capacity to produce OTA by Aspergillus section Nigri and Circumdati was investigated. A total of 50 corn samples for human consumption was collected in the south of Córdoba Province. The surface-disinfected method for mycobiota determination was used. The OTA detection was performed by HPLC. OTA production was tested in strains belonging to section Nigri and Circumdati. Statistical analysis demonstrated that the specie A. flavus was isolated in higher frequency (p<0.01) from corn kernels in DRBC and DG18 media. The percentage of corn kernels contaminated by A. niger var. niger was similar in DRBC and DG18 media. The frequency of grains contaminated by A. flavus and A. niger var. awamori was higher than A. niger var. niger and A. japonicus var. japonicus (p<0.01) in DG18 media. The other potentially ochratoxigenic species, A. ochraceus, was isolated between 5% and 10% of the corn kernels in DG18 and DRBC media, respectively. The OTA producing species P. verrucosum was not isolated. All samples of corn were OTA negative (<1 ng g⁻¹). Thirty strains (25%) of the black Aspergillus were OTA producers. From four strains of A. ochraceus isolated, only one produced OTA. Due to the storage variable conditions could not be adequate in this substrate, the presence of ochratoxigenic strains of section Nigri and OTA needs to be evaluated for a longer time to establish the toxicological risk for human beings. The contamination of stored corn kernels with A. flavus and Aspergillus section Nigri was significant.     

Keratinophilic fungi and other moulds associated with air-dust particles from Egypt

One-hundred and eleven species and three species varieties belonging to 39 genera were collected from 50 dust samples on the five media used at 28°C. Using the hair-baiting technique with horse hair, 10 species ofChrysosporium were isolated:C. asperatum, C. state ofArthroderma tuberculatum, C. indicum, C. inops, C. keratinophilum, C. merdarium, C. pannorum, C. queenslandicum, C. tropicum andC. xerophilum. True dermatophytes were isolated:Trichophyton verrucosum andTrichophyton sp. Also, numerous fungi tolerating high levels of cycloheximide were encountered, such as members ofAcremonium, Aspergillus andPenicillium. On plates of glucose or cellulose Czapek-Dox agar (free from sucrose) the most frequent fungi were:Alternaria alternata, Aspergillus flavus, A. flavus var.columnaris, A. fumigatus, A. niger, A. ochraceus, A. sydowii, A. terreus, Chaetomium globosum, Cladosporium herbarum, Emericella nidulans, Fusarium oxysporum, Mucor hiemalis, Penicillium chrysogenum, P. oxalicum, Scopulariopsis brevicaulis andUlocladium atrum. On plates of 50% sucrose or 10 and 20% NaCl-Czapek's agar, some interesting species were frequently encountered:Eurotium amstelodami, E. chevalieri, E. halophilicum, E. montevidensis, E. repens, E. rubrum andScopulariopsis halophilica. The isolated, fungi have been tested for osmophilicity and halophilicity, they showed different rates of growth on sucrose and sodium chloride-Czapek's medium of various osmotic potential.     

Antioxidant enzyme activity of filamentous fungi isolated from Livingston Island, Maritime Antarctica

From 18 soil samples taken in the vicinity of the permanent Bulgarian Antarctic base “St. Kliment Ohridski” (62°38′29″S, 60°21′53″W) on Livingston Island, 109 filamentous fungi were isolated on selective media. The most widespread fungal species were members of the genera Cladosporium, Geomyces, Penicillium and Aspergillus. Other species, already recorded in Antarctic environment, were also isolated: Lecanicillium muscarium, Epicoccum nigrum and Alternaria alternata. Thirty strains demonstrating good growth were screened for antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) that play an important role in the defense of aerobic organisms against oxidative stress, by converting reactive oxygen species into nontoxic molecules. Six of them showed high enzyme activity. The tested strains produced SOD with statistically significant higher activity at 15°C than at 30°C suggesting that this enzyme is cold-active. Such SOD could be useful in medicine and cosmetics. The best producer of cold-active SOD, Aspergillus glaucus 363, cultivated in bioreactors, demonstrated optimal growth temperature at 25°C and maximum enzyme activities at 25 and 30°C for SOD and CAT, respectively. The electrophoretical analysis showed that the fungus possesses Cu/Zn-SOD.     

The mycoflora of peanuts in Egypt

Twenty-six genera and Sixty-six species, in addition to two varieties of Aspergillus nidulans and one variety of Humicola grisea were isolated from 80 peanut seed (uncovered or covered within shell at the time of sampling) samples and 40 samples of shells collected from different places in Egypt.At 28 °C, the broadest spectrum of genera and species was recorded in the shells (21 genera and 53 species), followed by the naked seeds (16 genera and 38 species) and the covered seeds (17 genera and 33 species). Aspergillus and Penicillium were regularly the most common genera in the three substrates. A. niger, A. fumigatus, A. flavus A. terreus, A. ochraceus and P. funiculosum were generally the most common fungal species recovered from the three substrates. A. flavus was represented in 80%, 60% and 80% of the samples constituting 16.1%, 8.4% and 27.2% of the total count of fungi of the shells, covered and uncovered seeds respectively.At 45 °C, nine genera and sixteen species were collected of which Aspergillus fumigatus was extremely dominant in the three substrates, followed by A. terreus and A. niger. Penicillium was completely eliminated.     

Mycoflora and trichothecene toxins of paddy grains from Egypt

120 species and 38 genera were collected from 64 samples of paddy grains on glucose- and cellulose-Czapek's agar at 28 °C. The total count of glycophilic and cellulose-decomposing fungi fluctuated between 216–29760; and 124–11320 colonies/g paddy grains on the two media, respectively.On glucose agar, the most common species were Aspergillus niger, A. flavus, A. sydowi, A. terreus, A. fumigatus, A. ochraceus, Penicillium chrysogenum, P. corylophilum, Fusarium oxysporum, Alternaria alternata, Cladosporium cladosporioides, Trichoderma viride and Mucor racemosus.On cellulose agar with pH 5.5 & 8.0, the most prevalent fungi were Stachybotrys chartarum, S. bisybi, Aspergillus niger, A. flavus, Fusarium oxysporum, Alternaria alternata, Drechslera sativus and Acremonium strictum.Extracts from 64 paddy samples were tested against brine shrimp larvae (Artemis salina). Of these 9 displayed varying degrees of toxicity. Trichothecene-toxins were detected in the extracts of three paddy samples only. Diacetoxyscirpenol and T-2 toxin were detected in two samples and only T-2 toxin in the other.     

The mycoflora and toxicity of feedstuffs from a production plant in córdoba, Argentina

Feedstuffs used for poultry nutrition in Argentina were analyzed for fungal flora and natural incidence of mycotoxins. Survey of 120 samples of poultry feeds, taken from May 1998 to April 1999, showed the presence of 15 genera of filamentous fungi. The predominant genera wereFusarium spp. andPenicillium ssp., isolated in 67.5 % of the samples, followed byAspergillus spp. (57.5 %). Yeast, were significantly isolated from most of the samples. Species identification was carried down for the toxigenic genera. Fungal total counts of poultry feeds ranged from 2.0 × 10³ to 3.0 × 10⁵ CFU g^-1 The fungal total counts during two months of sampling, were slightly over the limit value of 1 × 105 CFU g^-1, which ensure the hygienic quality of the feed. Potentially toxicogenic species presented moderate mean colony counts. Many of the fungi isolated from poultry feeds are mycotoxin producers. Fumonisins had the highest incidence, and were found in 97 % of the analyzed samples followed by aflatoxin B₁ (46 %), zearalenone (18 %) and deoxynivalenol (6 %). On the co-occurrence of both carcinogenic mycotoxins, all of the FBs contaminated feed samples were co-contaminated with AFB₁. The results show the relevance of the samples screening for viable fungi propagules and the surveillance of their associated mycotoxins in poultry feeds.     

Incidence of seed-borne fungi and aflatoxins in Sudanese lentil seeds

Thirteen seed samples of lentil (Lens esculenta) were incubated on agar plate and moist filter papers (Moist Chambers) at 28 ± 2 °C for determination of the incidence of seed-borne fungi. Aflatoxins content of the seeds was measured using the bright greenish-yellow fluorescence test (BGYF) and thin-layer chromatography (TLC). Sixty-nine species and seven varieties, which belong to 24 genera of fungi, were isolated from this crop. Of these fungi, 51 species and two varieties are considered new for this crop, whereas seven genera and 13 species are new to the mycoflora of the Sudan. The genus Aspergillus (13 species and 6 varieties) which comprising 44% of the total colony count was the most prevalent genus followed by Rhizopus (2 species, 19%), Penicillium (6 species) and Fusarium (8 species) (12%), Chaetomium (3 species) and Cladosporium (5 species) (6%), where the 18 genera (1–4 species) showed very low level of incidence (19%). Of the possible pathogens of lentil plants, F. oxysporum the main cause of vascular wilt was recovered from seeds of this crop. Thin layer chromatographic analysis of chloroform extracts of 13 seed samples showed that only one samples was naturally contaminated with aflatoxins B₁, B₂, G₁ and G₂ (14.3 μg/kg). This revised version was published online in June 2006 with corrections to the Cover Date.     

Mycobiota and molecular detection of Aspergillus flavus and A. parasiticus aflatoxin contamination of Strawberry (Fragaria ananassa Duch.) fruits

Strawberry fungi were isolated from fresh fruits and juice on the two types of media (Sabouraud dextrose agar, SDA and potato-dextrose agar, PDA) at 28 °C. Nineteen fungal species belong to 12 genera were isolated from fruits and juice on both isolation media. The most common fungal genera and species were Aspergillus flavus, A. niger, Mucor racemosus, Neurospora crassa, Penicillium chrysogenum, Rhizopus stolonifer and Trichoderma harzianum. Twenty A. flavus and A. parasitics isolates were assayed for their abilities to produce aflatoxins. The concentration of aflatoxins ranged between 25.8–75.2 and 23.6–71.1 ng/ml at 350 and 365 nm, respectively. Among A. flavus and A. parasiticus strains tested, aflatoxin B contributed 30–60% of total isolates. However, G type contributed 85–90%. The R_f values of B₁, B₂, G₁ and G₂ were 0.79, 0.61, 0.44 and 0.32, respectively. High-performance liquid chromatography analysis of extracts revealed the presence of aflatoxins with variable levels.     

Fungi and aflatoxins associated with spices in the Sultanate of Oman

One hundred and five samples of seven spices (cumin, cinnamon, clove, black pepper, cardamom, ginger, and coriander) were purchased from five popular companies in the Sultanate of Oman. The spices were surveyed for the mycoflora and aflatoxins. Twenty fungal species were isolated in which Aspergillus flavus , A. niger . Penicillium , Rhizopus , and Syncephalastrum racemosum were the most dominant. When colony forming units per gm (cfu/gm) of fungi were compared, significant differences were found among spices and companies. Of the seven spices studied, clove was found to be the least contaminated, while cumin was the most contaminated. None of the 15 selected samples of the spices contaminated by A. flavus were found to contain aflatoxins. Nevertheless, nine isolates (45%) of the twenty A. flavus strains screened for aflatoxins were aflatoxigenic. The moisture content of most of the spices was below the maximum standard limit. The results showed that the spices were contaminated by some fungi that might constitute health hazards for humans. This revised version was published online in June 2006 with corrections to the Cover Date.     

Survival of Micromycetes and Actinobacteria under Conditions of Long-Term Natural Cryopreservation

Almost all of the investigated samples of the Arctic and Antarctic permafrost sediments of different genesis with ages from 5–10 thousand to 2–3 million years were found to contain viable micromycete and bacterial cells. The maximum amounts of viable cells of fungi (up to 10⁴CFU/g air-dried sample) and bacteria (up to 10⁷–10⁹CFU/g air-dried sample) were present in fine peaty sediment samples taken from different depths. The identified micromycetes belonged to more than 20 genera of the divisions Basidiomycota, Ascomycota, and Zygomycota, and some represented mitosporic fungi. Thawing the samples at 35 and 52°C allowed the number of detected fungal genera to be increased by more than 30%. Aerobic heterotrophic prokaryotes were dominated by coryneform, nocardioform, and spore-forming microorganisms of the order Actinomycetales.Analysis of the isolated fungi and actinomycetes showed that most of them originated from the microbial communities of ancient terrestrial biocenoses.