Fungi Isolated from Flue-cured Tobacco at Time of Sale and After Storage

The fungi isolated from 100 samples of flue-cured tobacco from 12 markets in 2 tobacco belts comprised 11 genera, including 10 species of Aspergillus. The mean percentage per sample isolated from 62 samples of tobacco from Middle Belt markets was Alternaria, 40.6%; Aspergillus niger, 47.8%; Aspergillus repens, 38.0%; and Penicillium, 25.8%. The mean percentage per sample isolated from 38 samples of tobacco from Old Belt markets was Alternaria, 74.0%; Penicillium, 52.5%; Aspergillus repens, 38.0%; and Aspergillus ruber, 36.2%. Damaged (74 samples) and nondamaged (26 samples) stored tobacco yielded species of six genera of fungi, including eight species of Aspergillus. Species of Aspergillus and Penicillium were commonly isolated from both damaged and nondamaged tobacco, whereas species of Alternaria, Cladosporium, Fusarium, and Rhizopus were isoalted more frequently from nondamaged tobacco. The fungi that occurred in the highest population in damaged tobacco were Aspergillus repens, A. niger, A. ruber, and Penicillium species.     

Mycoflora of Argentinian corn harvested in the main production area in 1990

Corn (Zea mays) is the main cereal produced in and exported from Argentina. The risk of contamination by mycotoxins is related to the mycoflora associated with the corn kernels. This paper reports on the identification of internal and external mycoflora of corn kernels harvested in the main production area in Argentina in 1990. A mycological survey was carried out on 178 corn samples, from five locations in that area and the isolation frequency and relative density of the prevalent fungal genera compared. GenusFusarium was the most prevalent component of the internal seedborne mycoflora in the five locations.Penicillium was prevalent in all locations, taking into account the frequency. However, this genus was predominant only in two locations, when the relative density was considered. The predominantFusarium wasF. moniliforme and the most frequently isolated species ofAlternaria, Aspergillus andPenicillium wereA. alternata, A. flavus andP. decumbens, respectively.Diplodia species were not isolated from any of the samples.     

Investigation of stored wheat mycoflora,reporting the<Emphasis Type="Italic">Fusarium</Emphasis> cf.<Emphasis Type="Italic">langsethiae</Emphasis> in three provinces of Iran during 2007

Wheat is the most important cereal produced in Iran. A mycological survey was carried out for the first time, on the stored wheat samples in Tehran, East Azarbayejan and Mazandaran provinces in 2007. Exogenous and endogenous fungi, were isolated by the method of flotation with Malachite green agar (MGA 0.25) and Freeze blotter techniques respectively. In this study, 46 species belonging to 23 different genera were isolated.Cladosporium spp. (57.1–89.2%) andAlternaria spp. (82.4–100%) species were the predominant fungal species identified as endogenous mycoflora. The predominant exogenous fungi werePenicillium spp. (78.4–92.8%) andAspergillus spp. (71.4–85.7%) species.Fusarium proliferatum was the most prevalent species ofFusarium isolates.Aspergillus niger (39.4%) andAspergillus flavus (36.7%) were the predominantAspergillus species identified as exogenous mycoflora.Aspergillus flavus (26.6%) was the predominantAspergillus species identified as endogenous mycoflora. Flotation method with MGA 0.25 recommended for isolating of hyaline fungi from wheat cereals. In this study one isolate fromFusarium species was isolated on the basis of morphology and ribosomal internal transcribed spacer classified asFusarium langsethiae but on the basis of partial translation elongation factor-1alpha gene grouped withFusarium sporotrichioides. To our knowledge, this is the first report aboutF. cf.langsethiae in Iran and Asia.     

Survey of the mycoflora of barley grains in Egypt

The fungi of 40 samples of barley grains collected from Upper Egypt were estimated using the dilution- and the grain-plate methods. 33 genera and 109 species, in addition to 2 varieties of Aspergillus nidulans, one variety of Aspergillus flavus and one variety of Penicillium cyclopium were identified. The spectrum of fungal genera and species collected by the grain-plate method (29 genera and 94 species+3 varieties) was broader than by the dilutionplate method (27 genera and 71 species+3 varieties), but the order of frequency of occurrence of fungal genera and species was basically similar in the two methods. The most frequent genera were Aspergillus (25 species+3 varieties), Penicillium (32 species+1 variety), Rhizopus (2 species), Alternaria (2 species), Fusarium (6 species) and Drechslera (5 species) and were represented (as assayed by the grainplate method) in 100 %, 77.5 %, 85 % 62,5 %, 60 % and 52.5 % of the samples constituting 60.4 %, 10.2 %, 7.7 %, 3.7 %, 5 % and 2.3 % of the gross total count of fungi respectively. From the preceding genera A. niger, A. fumigatus, A. flavus, A. sydowii, P. notatum, P. citrinum, F. oxysporum, F. moniliforme, F. solani, R. stolonifer, A. alternata and D. spicifera were the most frequent.     

Mycoflora and mycotoxin contamination of Roundup Ready soybean harvested in the Pampean Region, Argentina

A total of 89 freshly harvested soybean seed samples (Roundup Ready [transgenic] soybean cultivars) from the 2010/2011 crop season were collected from five locations in the Northern Pampean Region II, Argentina. These samples were analyzed for internal mycoflora, toxin production of isolated fungi, and for a range of mycotoxins. Mycotoxin analysis of aflatoxins (AFs), zearalenone (ZEA), fumonisins (FBs) and ochratoxin A (OTA) was done by HPLC-FLD (high performance liquid chromatography with postcolumn fluorescence derivatization), alternariol and alternariol monomethyl ether with HPLC-UV (HPLC with UV detection), trichothecenes (deoxynivalenol, nivalenol, T-2 toxin, HT-2 toxin, fusarenon X, 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol were analyzed by GC-ECD (gas chromatography with electron capture detector). Fungal colonization was more frequently found for samples from América, Saladillo and Trenque Lauquen than for samples from General Villegas and Trenel; a total of 1,401 fungal isolates were obtained from the soybean seeds. The most commonly identified fungal genera were Alternaria, Sclerotinia, Chaetomium, Cladosporium, Aspergillus, Penicillium, Phomopsis and Fusarium. Alternaria alternata, A.tenuissima, Aspergillus flavus, Penicillium citrinum, Fusarium verticillioides and F.semitectum were the predominant toxigenic fungal species. Mycotoxin production was confirmed for several isolates of toxigenic species, including Aspergillus flavus, A. parasiticus, Alternaria alternata, A.tenuissima, Fusarium graminearum, F semitectum and F. verticillioides. In particular, the percentage of mycotoxigenic Alternaria alternata (100 %), A.tenuissima (95 %) and aflatoxigenic strains of A. flavus (57 %) were remarkably high. Although none of the mycotoxins, AFs, ZEA, FBs, trichothecenes and OTA, were directly detected in samples of soybean seeds, the frequent presence of toxigenic fungal species indicates the risk of multiple mycotoxin contamination.     

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.     

Mycobiota and mycotoxins in fermented feed,wheat grains and corn grains in Southeastern Buenos Aires Province,Argentina

Wheat (as bran) and corn (as dry grain or fermented feed) are main ingredients of feedstuffs used in local cattle and pig farms in the South of the Buenos Aires Province (Argentina). Therefore, determining mycobiota and mycotoxins in wheat and corn is of prime importance for developing feed management techniques to optimise animal production and to minimize toxicity. Then, a mycological survey was carried out in the Southeastern part of the Buenos Aires Province, in order to identify the mycobiota and the main mycotoxins present in fermented feed, wheat grain and corn grain samples. Samples were cultured for fungal quantification, isolation and identification, and analysed for deoxynivalenol (DON), zearalenone (ZEA), T-2 toxin and aflatoxins (AFLA). Penicillium (74%), Aspergillus (32%) and Scopulariopsis (21%) were the prevalent genera in fermented feed. Penicillium (70%), Fusarium (47%) and Aspergillus (34%) were the most frequent fungi isolated from corn. Penicillium (42%), Fusarium (27%) and Alternaria (25%) were the most frequently recovered genera from wheat. DON was detected in 59% of the corn samples, in 45% of the wheat samples and in 38% of the silage samples. ZEA was detected in 36% of the corn samples, in 49% of the wheat samples and in 16% of the silage samples. T-2 toxin and aflatoxin B1 were each detected in 4% of the corn samples. Eighteen percent of the fermented feed samples showed T-2 contamination. Fermented feed and wheat samples were negative for AFLA.     

Fungi associated with rice at entre rios province, Argentina. toxigenic capacity offusarium graminearum andmicrodochium nivale isolates

A mycological survey was carried out on rice samples harvested in 1997 and 1998 from Entre Ríos province, belonging to the main production area of Argentina. The relative density and isolation frequency of the prevalent fungi were statistically compared between locations and harvest seasons. The genusAlternaria was the most prevalent component of the internal seedborne mycoflora in the two harvest seasons. Fungi belonging to the generaPhoma, Fusarium, Microdochium, Penicillium andAspergillus were also isolated. The predominant species of these genera wereAlternaria alternata, Phoma glomerata, Fusarium graminearum, Microdochium nivale, Penicillium citrinum and Aspergillus flavus.     

Diversity and Drug Resistance of Filamentous Fungi Isolated from the Fresh Raspberries

Fungi are one of the most widely distributed microorganisms in the environment, including food such as fruits, vegetables and other crops, posing a potential threat to food safety and human health. The aim of this study was to determine the diversity, intensity and drug resistance of potentially pathogenic filamentous fungi isolated from the fresh raspberries (Rubus idaeus L.). A total of 50 strains belonging to genera Fusarium, Cladosporium, Alternaria, Penicillium, Mucor, Rhizopus, Aspergillus and Acremonium were tested for drug resistance against 11 antifungals by disc diffusion and gradient strips methods. The average mycological contamination in the examined samples of raspberries amounted to 4.34 log CFU/g. The Cladosporium was isolated from all tested samples, followed by Alternaria and Fusarium with a frequency of 61% and 34%, respectively. The highest level of drug resistance was observed for Acremonium genera and Fusarium strains recorded a wide variation in drug resistance as revealed by susceptibility with amphotericin B and voriconzole with MICs ranged from 0.5–4 µg/ml and posaconazole with MICs ranging from 3–8 µg/ml. All fungal strains showed 100% resistance to caspofungin, fluconazole and flucytosine with both the methods, and 100% resistance to micafungin and anidulafungin in the gradient strip method.     

Mycoflora and naturally occurring mycotoxins in poultry feeds in Argentina

The purpose of this work was to determine the mycoflora and mycotoxins natural incidence in poultry feeds from 2 factories in Río Cuarto, Córdoba. One hundred and thirty samples were taken from May/1996 to May/1997. The most dominant species isolated of poultry feed samples belonged to the genera Aspergillus spp 85% and Fusarium spp 70%. From Aspergillus genus eleven species were identified and A. flavus was the most frequent. Nine species were identified from the Fusarium genus and the predominant was F. moniliforme. Penicillium ranked third in the number of isolated cases. From this genus twelve species were collected of which P. brevicompactum (15%), P. restrictum (14%) and P. purpurogenum (12%) were the most common.The most significant mycotoxin from poultry feeds was aflatoxin B₁ (AFB₁) found in 48% of the samples, with levels ranging from 10 to 123 ng/g. For zearalenone (ZEA) the levels were 327 to 5,850 ng/g and DON was not detected from the samples. Due to the fact that in Argentina there is little information about this topic, these data on poultry feeds in our region would be of worldwide interest.This revised version was published online in October 2005 with corrections to the Cover Date.     

Seasonal variation of fungal spore populations in the atmosphere of Zarqa area,Jordan

Fungi are among the most important aeroallergens. The aim of this study was to provide aeromycological baseline information about Zarqa area, Jordan, for the first time. During the entire survey and from 170 settle plate exposures, a total of 735 mould- and 274 yeast colony-forming units (CFU) were collected. mould colonies were assigned to 35 genera and 59 species. The highest abundance was attributed to Cladosporium with a percentage of 29.1% of the total colony count followed by Fusarium 20%, Alternaria 7.7%, Ulocladium 6.5% Penicillium 4.2% and then Aspergillus 3.6%. Cladosporium showed one peak in October while Aspergillus and Penicillium peaked in September. Fusarium peaked in May and August and Alternaria in March and July. However, Ulocladium showed almost the same abundance from March to June and then in September and December. A double peak in total colony count and fungal diversity was found, the first was in March and the second in November–December. Significant differences of total fungal colony counts favouring the 1500 hours collection time over that at 1000 hours were found between November and January while no such significant differences were obtained between February and April. Our results also indicated differences in species composition between the two periods in different months. Spores of the three abundant species, Cladosporium, Fusarium and Alternaria, might favour high release in the 1500 hours period during winter while having a similar incidence during the two periods in the spring months.     

Aspergillus and other moulds in the air of Kuwait

A one-year survey was carried out to study the aerial prevalence of Aspergillus species and other moulds in the outdoor and indoor environments of Kuwait. Petri plates containing rose-Bengal medium were exposed for 20 minutes twice a month using a six-stage Andersen air sampler at the pre-determined sites. The exposed plates were incubated at 28 °C ± 1 °C up to 5 days and colonies were enumerated and identified by colonial and microscopic morphology. The data revealed that Aspergillus species were the predominant component (27.7%) of the outdoor aerospora of Kuwait and A. fumigatus alone accounted for 21.3% of the total aspergilli. In contrast, Cladosporium species formed the major component of the indoor aerospora (22.8%), followed by Aspergillus species (20.9%), Penicillium species (14.6%), and Bipolaris species (10.6%). A comparison of the fungi recorded in the outdoor and in the indoor air revealed that Aspergillus, Alternaria and Fusarium were significantly higher in the outdoor environment, whereas Cladosporium, Penicillium, and Bipolaris were significantly higher in the indoor environment. The relative prevalence of Aspergillus species and other moulds in the outdoor and indoor air of Kuwait was as follows: A. fumigatus 5.9 and 9.8%, A. flavus 4.9 and 3.9%, other aspergilli 16.8 and 7.0%, Alternaria species 19.8 and 7.9%, Cladosporium species 13.7 and 22.8%, Penicillium species 7.6 and 14.6%, and other moulds 31.2 and 34.1%, respectively. During the study, 25 different genera were identified, indicating a wide diversity in the spectrum of local fungal aerospora. The study provides useful information on the prevalence of allergenic fungi in the outdoor and indoor environments of Kuwait. This revised version was published online in June 2006 with corrections to the Cover Date.     

A Brevibacillus sp. antagonistic to mycotoxigenic Fusarium spp.

Antagonistic microbes were isolated from soils to control mycotoxin contamination of cereals by limiting the growth of mycotoxigenic Fusarium species. In total, 341 bacterial isolates were examined for antifungal activity against eight mycotoxigenic Fusarium species using dual culture assays. The screening identified 11 isolates that inhibited mycelial growth of all Fusarium species tested. The culture filtrates of 2 of the 11 isolates completely inhibited germination of conidia up to 21 days of incubation. These two isolates exhibited identical activity toward the fungi tested and were identified as Brevibacillus spp. based on 16S rRNA sequence homology. The most closely related species based on phylogenetic analysis was Brevibacillus reuszeri. Additional dual culturing using further fungal species showed that the antagonistic Brevibacillus inhibited the growth of most Fusarium species tested (39 of 46 species), two Epicoccum spp., one Alternaria sp., three Aspergillus spp. (3 of 11), and three Penicillium spp. (3 of 8). The in vivo assay was performed to test the efficacy of antagonistic Brevibacillus isolates on maize ears and revealed that the application of microbes suppressed ear rot (ANOVA, p = 0.0020). This Brevibacillus sp. may be an antagonist of the majority of Fusarium species, including mycotoxigenic species.     

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.     

Characterization of fungi transferred by dust storms in Kuwait and their plant pathogenicity

Dust storms carry large amounts of plant detritus and microorganisms that may cause diseases in humans, animals or plants. These storms are frequent in Kuwait throughout the year. This research was conducted to identify the fungal species carried by the dust storms in Kuwait, originating from the northwesterly direction, with emphasis on plant pathogens. Fungi were isolated from settled dust samples and identified using established microbiological and molecular approaches. Fungal isolates identified as Fusarium oxysporum from settled dust were examined for pathogenicity using a number of crop plants. In total, 17 genera of fungi were identified in the dust samples. These fungi included plant pathogens or facultative plant parasites that were transported in the dust storms as viable propagules. The most common dust-carried fungi belonged to the genera Fusarium, Alternaria, Ulocladium, Phoma, Aspergillus, Acremonium and Penicillium. The F. oxysporum isolates that had been characterized by partial 18S rRNA gene sequencing were pathogenic, causing root and stem rot in tomato, bean and cucumber, but not squash.     

Environmental Isolates of Fungi from Aquarium Pools Housing Killer Whales (Orcinus orca)

Systemic mycoses in killer whales (Orcinus orca) are rare diseases, but have been reported. Two killer whales died by fungal infections at the Port of Nagoya Public Aquarium in Japan. In this study, the fungal flora of the pool environment at the aquarium was characterized. Alternaria spp., Aspergillus spp. (A. fumigatus, A. niger, A. versicolor), Fusarium spp. and Penicillium spp. were isolated from the air and the pool surroundings. The other isolates were identified as fungal species non-pathogenic for mammals. However, the species of fungi isolated from the environmental samples in this study were not the same as those isolated from the cases of disease in killer whales previously reported.     

Contribution to the fungal flora of cereal grains in Egypt

30 genera and 77 species, in addition to 4 varieties were isolated from 25 samples of each of barley, wheat, maize and sorghum grains collected from different places in Egypt. The broadest spectrum of genera and species was recorded in wheat (25 genera and 59 species + 4 varieties) followed by barley (21 genera and 52 species + 2 varieties), sorghum (14 genera and 33 species + 2 varieties) and maize grains (11 genera and 29 species + 2 varieties). Aspergillus, Fusarium, Penicillium and Rhizopus were the most common genera in the four grains, except maize where Penicillium emerged in low frequency. Aspergillus was the main component of the fungal flora of the four grains and contributed 79–94.4% of the gross total count of fungi. From the preceding genera A. niger, A. flavus, A. fumigatus, F. oxysporum, P. chrysogenum, P. corylophilum, P. notatum and R. stolonifer were the most frequent.     

Seed-borne fungi in domestic bird feed in Saudi Arabia

Seed-borne fungi from seeds of barley, canary, maize, oats, rice, rapeseed, red millet, sunflower, white millet and yellow millet were investigated. These seeds are available in the market as bird feed, supplied by companies in various countries as healthy and nutritious food for domestic birds. A total of 81 fungal species were isolated. The genusAspergillus was predominant with 23 species, followed byPenicillium with 18,Ulocladium with 7,Alternaria with 6 andChaetomium, Curvularia andDrechslera with 5 species each. Other genera isolated wereAureobasidium, Cladosporium, Fusarium, Mucor, Rhizopus andSyncephalastrum.     

Microfungal Contaminants on Mobile Phones of Health Services Vocational School Students in Marmaris,Turkey

In this study, it was aimed to determine microfungi on mobile phones. Totally, 50 mobile phones were used belonging to Health Services Vocational School students. The samples were taken by swabbing the screen and keys of mobile phones using moistened sterile swab sticks. A total of 24 different microfungal species were obtained belonging to Alternaria, Aspergillus, Cladosporium, Geotrichum, Penicillium, Phoma, Rhinocladiella, Scopulariopsis, Trichoderma, and Trichophyton genera. The genera of microfungi most abundant in terms of the number of species on the mobile phones were Aspergillus, Cladosporium, and Penicillium. Numerically, Cladosporium was found as the most abundant on the mobile phones. Cladosporium herbarum colonies were highest in number, followed by Cladosporium sphaerospermum, and Penicillium verrucosum var. cyclopium. When percentages of each species present on the mobile phones were considered, C. herbarum and C. sphaerospermum were the most common. There was a great similarity between the dominant microfungi isolated from mobile phones and dominant microfungi obtained from studies of atmospheric microfungi in Turkey.     

Mycoflora and mycotoxins of peanut (Arachis hypogaea L.) seeds in Egypt. III. Cellulose-decomposing and mycotoxin-producing fungi

From 40 peanut seed samples collected in Egypt, forty-three species and one variety of fungi, belonging to 16 genera, were collected. The most dominant genera were Aspergillus (11 species + one variety), Penicillium (11 species) and Fusarium (4 species). From the preceding genera A. fumigatus, A. flavus, A. niger, P. chrysogenum and F. oxysporum were the most frequent species.Forty-nine isolates belonging to 12 species and one variety were tested for production of mycotoxins, after growth on liquid medium containing two carbon sources (sucrose or cellulose). Thin layer chromatographic analysis revealed that the quality and quantity of mycotoxins was higher on sucrose than cellulose. Mycotoxins identified were aflatoxins B₁, B₂, G₁ & G₂, citrinin; fumagillin; diacetoxyscirpenol T-2 toxin; satratoxin H; and zearalenone.