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Fungal otitis externa, an infection of the external auditory canal caused by molds and yeasts, accounts for approximately 10–20% of ear canal infections accompanying high recurrence. The purpose of the current study was to assess the pattern of etiological agents of otomycosis and resistance profile as well as the rate of tympanic membrane perforation. A total of 1040 patients with symptoms of fungal otitis externa, in a period of two years, were investigated. The mycological tests revealed the presence of different fungi in 237 ears (22.8%). Fungal otitis was more related to filamentous fungi of the species Aspergillus flavus (54.43%), A. tubingensis (10.97%), and A. niger (8.86%), followed by yeasts, Candida orthopsilosis (7.59%), C. albicans (6.75%), and C. parapsilosis (5.06%). Tympanic membrane perforation rate was found to be 6.75% and was more common with otomycosis caused by A. flavus, A. tubingensis and C. albicans. In antifungal susceptibility tests, all tested drugs showed generally good activity against most isolates of molds and yeasts, while tolnaftate, clotrimazole, nystatin, and terbinafine had lowest effects. We found that among Aspergillus isolates, one A. niger isolate was resistant to voriconazole, and one A. flavus isolate was resistant to amphotericin B. Furthermore, among Candida species, three isolates of C. orthopsilosis showed high MIC values to fluconazole, two C. albicans isolates were considered fluconazole resistant and one isolate of C. parapsilosis was resistant to caspofungin and 3 isolates were resistant to fluconazole. Regarding the existence of the cases with perforated tympanic membrane and emerging species causing fungal otitis in the current report, the importance of the early physical examination, precise molecular identification, and the antifungal susceptibility evaluation is highlighted. [ABSTRACT FROM AUTHOR]

Lichtheimia corymbifera and Aspergillus flavus pulmonary coinfection has been rarely reported in immune-competent patients. We report case of a young male who presented with clinical features of pulmonary-renal syndrome and was later diagnosed to have bilateral polymicrobial fungal lung infection. [ABSTRACT FROM AUTHOR]

We investigated the clinical and mycological characteristics of four cases of mycotic keratitis caused by Aspergillus flavus that occurred from July 2014 to May 2015 at Çukurova University Hospital, Adana, Turkey. In a 10-month period, a total of 64 corneal smear/scrapings were examined from patients with suspected mycotic keratitis. Fungal cultures were positive in six of these patients, indicating a 9.4% incidence of mycotic keratitis in this region, including four cases of A. flavus and two cases of Fusarium spp. The predisposing factors, clinical presentation, and success of the therapeutic approaches were further evaluated. For all cases, topical voriconazole was the first choice of treatment. Surgical procedures were required to control infection in 3 of the 4 cases, including intrastromal voriconazole injection for two cases and keratoplasty for one case. Predisposing factors included trauma (two cases, 50%), contact lens use (one case, 25%), and previous ocular surgery (one case, 25%). The clinical presentations also differed, including a well-limited ulcer (one case), an ulcer with an irregular feathery margin (one case), and ulcers with satellite lesions (two cases). The mean duration between the time of presentation and definitive diagnosis by culture was 14 days (8-25 days). We observed that A. flavus keratitis can present with different underlying factors and clinical conditions. A combination of antifungal therapy and supportive surgical intervention may resolve infections caused by A. flavus in the cornea. [ABSTRACT FROM AUTHOR]

Aspergillus flavus is the second most leading cause of aspergillosis. The ability of A. flavus to adapt within the host environment is crtical for its colonization. Onset of germination of conidia is one of the crucial events; thus, in order to gain insight into A. flavus molecular adaptation while germination, protein profile of A. flavus was obtained. Approximately 82 % of conidia showed germination at 7 h; thus, samples were collected followed by protein extraction and subjected to nLC-Q-TOF mass spectrometer. Q-TOF data were analysed using Protein Lynx Global Services (PLGS 2.2.5) software. A total of 416 proteins were identified from UniProt Aspergillus species database. Orthologues of A. flavus was observed in A. fumigatus, A. niger, A. terreus, A. oryzae, etc. Proteins were further analysed in NCBI database, which showed that 27 proteins of A. flavus are not reported in UniProt and NCBI database. Functional characterization of proteins resulted majorly to cell wall synthesis and degradation, metabolisms (carbohydrate and amino acid metabolism), protein synthesis and degradation. Proteins/enzymes associated with aflatoxin biosynthesis were observed. We also observed Dicer-like proteins 1, 2 and autophagy-related proteins 2, 9, 18, 13, 11, 22. Expression of protein/enzymes associated with MAPK signalling pathway suggests their role during the germination process. Overall, the data present a catalogue of proteins/enzymes involved in the germination of A. flavus conidia and could also be applied to other Aspergillus species. [ABSTRACT FROM AUTHOR]

Aspergillus flavus is the most common species associated with invasive aspergillosis in Tunisia. The molecular epidemiology of the species is poorly documented. We used five highly discriminative microsatellite markers for the genotyping of clinical and hospital environmental A. flavus strains to assess whether IA could be hospital-acquired in the onco-hematology unit of the Farhat Hached teaching hospital of Sousse, Tunisia. The genotyping of 18 clinical isolates, collected from sputa of 17 acute leukemia patients, and 81 isolates, collected in these patients' hospital environment and food, identified 57 isolates that were grouped in 10 clones, each of them including 2-17 isolates. The remaining 42 isolates showed a unique genotype. Two main transmission scenarios were observed: (1) the same clone was isolated from different patients; (2) the same clone was isolated from a patient, its hospital environment and/or food. These findings strongly suggest the occurrence of hospital-acquired A. flavus infection/colonization in the investigated onco-hematology unit. [ABSTRACT FROM AUTHOR]

Aspergillosis is one of the most frequent mycosis affecting avian species. Here is reported an outbreak of aspergillosis affecting 60-day-old white Pekin mallards ( Anas platyrhynchos). About 10 % of animals in a lot of 200 mallards from a commercial husbandry presented respiratory disorders and skin lesions at slaughter. Three out of 13 animals sent to diagnosis showed, simultaneously, airsacculitis, lung and liver presenting white nodules with variable diameters and elevated, yellowish brown, crusted, multifocal skin lesions located at the base of the feather follicles in the breast. Histopathological examination of lung and liver samples revealed nodules of different sizes with small areas of necrosis surrounded by intense granulomatous inflammation and the presence of fungal hyphae. The skin samples showed dermatitis surrounding a severe necrotizing folliculitis, associated with fungal hyphae. Mycological evaluation of tissues allowed the isolation of Aspergillus fumigatus from the skin samples and Aspergillus flavus from lungs and liver samples. The application of quicklime (CaO) in the litter as part of the disinfection procedures could have contributed to the development of skin lesion in the mallards, predisposing the fungal installation in the damaged site. The occurrence of cutaneous lesions associated with A. fumigatus is a rare manifestation of aspergillosis in birds, and this appears to be the first case reported in white Pekin mallards. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the efficacy of topical application of a liposomal formulation of itraconazole for the treatment of experimental keratitis with endophthalmitis caused by Aspergillus flavus. The liposomes were obtained by the lipid film hydration method followed by sonication. Adult female Wistar rats (weighing 200-220 g) were immunosuppressed by intraperitoneal injection of 150 mg/kg of cyclophosphamide 3 days before infection by exposure to the fungus A. flavus (10 spores/ml). Forty-eight hours later, the animals were treated with the liposomal formulation. For comparison, one group of animals ( n = 6) was treated with the same drug not encapsulated. At the end of the experiment, the animals were evaluated for clinical signs and number of colony forming units (CFU/g), along with direct microscopic examination. The results indicated that the liposomal formulation of itraconazole has better antifungal activity than the unencapsulated drug in the treatment of fungal keratitis with endophthalmitis caused experimentally by A. flavus in Wistar rats. [ABSTRACT FROM AUTHOR]

Objectives: Invasive pulmonary aspergillosis (IPA) caused by Aspergillus fumigatus, Aspergillus flavus, or Aspergillus niger is associated with high mortality. We evaluated the efficacy and compared the therapeutic effect differences of voriconazole (VRC) in combination with caspofungin (CAS) in transiently neutropenic rats infected by A. fumigatus, A. flavus, or A. niger. Methods: Treatment groups consisted of VRC (10 mg/kg q12 h) monotherapy, CAS (1 mg/kg/day) monotherapy, combination of VRC (10 mg/kg q12 h) + CAS (1 mg/kg/day), and no drug for 10 consecutive days. The efficacy and the difference in the treatments were evaluated through prolongation of survival, reduction in serum galactomannan levels and residual fungal burden, and histological studies. Results: For all the strains, the combination of VRC and CAS led to significant prolongation in survival ( P < 0.05) and reduction in residual fungal burden ( P < 0.05) compared with CAS alone, and decrease in serum galactomannan levels ( P < 0.05) compared with either agent alone. The survival in the combined therapy groups was significantly improved compared to VRC monotherapy for the strains of A. flavus and A. niger ( P < 0.05), but no significant difference for the strains of A. fumigatus ( P > 0.05). Conclusions: Combination of VRC and CAS was synergistic in IPA by A. flavus and A. niger, but small efficacy benefits in IPA by A. fumigatus. [ABSTRACT FROM AUTHOR]

Simple sequence repeat (SSR) markers were developed from Aspergillus flavus expressed sequence tag (EST) database to conduct an analysis of genetic relationships of Aspergillus isolates from numerous host species and geographical regions, but primarily from the United States. Twenty-nine primers were designed from 362 tri-nucleotide EST-SSR sequences. Eighteen polymorphic loci were used to genotype 96 Aspergillus species isolates. The number of alleles detected per locus ranged from 2 to 24 with a mean of 8.2 alleles. Haploid diversity ranged from 0.28 to 0.91. Genetic distance matrix was used to perform principal coordinates analysis (PCA) and to generate dendrograms using unweighted pair group method with arithmetic mean (UPGMA). Two principal coordinates explained more than 75 % of the total variation among the isolates. One clade was identified for A. flavus isolates ( n = 87) with the other Aspergillus species ( n = 7) using PCA, but five distinct clusters were present when the others taxa were excluded from the analysis. Six groups were noted when the EST-SSR data were compared using UPGMA. However, the latter PCA or UPGMA comparison resulted in no direct associations with host species, geographical region or aflatoxin production. Furthermore, there was no direct correlation to visible morphological features such as sclerotial types. The isolates from Mississippi Delta region, which contained the largest percentage of isolates, did not show any unusual clustering except for isolates K32, K55, and 199. Further studies of these three isolates are warranted to evaluate their pathogenicity, aflatoxin production potential, additional gene sequences (e.g., RPB2), and morphological comparisons. [ABSTRACT FROM AUTHOR]

To study the effects of and interactions among feed types, seasons, and agroecological zones on the total fungal viable count and aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1), and G2 (AFG2) production in poultry feed, an experiment was conducted using three-factorial design. A total of 216 samples of poultry feed ingredients, viz. maize, wheat, rice, cotton seed meal (CSM), and finished products, that is, starter and finisher broilers' rations, were collected from Peshawar, Swat, and D. I. Khan districts of Khyber Pakhtunkhwa, Pakistan, during the winter, spring, summer, and autumn seasons of the year 2007/2008. Analysis of variance showed that there was a complex interaction among all these factors and that this influenced the total fungal viable count and relative concentrations of the aflatoxins produced. Minimum total culturable fungi (6.43 × 10 CFUs/g) were counted in CSM from D. I. Khan region in winter season while maximum (26.68 × 10 CFUs/g) in starter ration from Peshawar region in summer. Maximum concentrations of AFB1 (191.65 ng/g), AFB2 (86.85 ng/g), and AFG2 (89.90 ng/g) were examined during the summer season whereas the concentration of AFG1 was maximum (167.82 ng/g) in autumn in finisher ration from Peshawar region. Minimum aflatoxins were produced in the winter season across all the three agroecological zones. [ABSTRACT FROM AUTHOR]

The efficacy of the combination of anidulafungin (AFG) at 1 mg/kg plus voriconazole (VRC) at 25 mg/kg was evaluated in a murine model of disseminated infection by Aspergillus flavus using three isolates previously tested in vitro. All the combinations showed indifferent in vitro interaction with the exception of one, which showed synergy. In general, the combined treatment prolonged the survival and reduced the fungal load in comparison with AFG alone, and only in a few cases, it improved the results of the VRC monotherapy. The combination of the two drugs and VRC alone reduced the galactomannan levels in serum in comparison with the control group. [ABSTRACT FROM AUTHOR]

Aspergillus flavus K49 secreted at least two xylanase activities when grown on a medium containing larch (wood) xylan as a sole carbon source. Enzyme activity was assayed using an agar medium containing Remazol Brilliant Blue R conjugated oat spelt xylan as substrate. Crude enzyme preparations were inhibited by Hg, with an ED of 17.5 mM and maximum inhibition of 83% at 50 mM. A concentrated sample of A. flavus K49 xylanase preparation was subjected to gel filtration chromatography on a P-30 column. A small protein peak coinciding with the major peak of xylanase activity was separated from the other secreted fungal proteins. An additional peak of xylanase activity was observed in fractions containing multiple fungal proteins. Analysis by denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of fractions containing the smaller molecular weight xylanase revealed a major and minor protein band in the vicinity of 14 kD. Analysis of these same fractions by acidic native PAGE revealed a single band. Confirmation of identity for the isolated xylanase was provided by isolation of a protein band from a SDS-PAGE gel, followed by trypsin digestion/analysis by tandem mass spectrometry. Comparison of the peptide library derived from this protein band with sequence data from the A. oryzae genomic data base provided a solid match with an endo-1,4-β-xylanase, XlnA. This identification is consistent with a low molecular weight protein associated with the major xylanolytic activity. XlnA may be a highly mobile (diffusible), plant wall hemicellulose degrading factor with significant activity during plant infection. [ABSTRACT FROM AUTHOR]

survey was carried out to detect the presence of aflatoxin B in 60 duplicated samples (120 samples) of peanuts butter purchased from the local markets and other traditionally prepared and distributed by the street sellers in Khartoum state, Sudan. AflaTest-P affinity column was used to extract the toxin from the samples, and the concentration was measured by calibrated Vicam fluorometer. Aflatoxin B was detected at variable levels in 100% of the screened samples. Traditionally prepared samples showed the highest incidence of aflatoxin B which is above the internationally regulated tolerance levels (5-20 ppb). The means and the ranges of the aflatoxin B recovered were as follows: 63.9 ppb (29-128 ppb), 54.5 ppb (21-131 ppb) and 101 ppb (17-170 ppb) for samples collected from Khartoum, Khartoum North and Omdurman areas, respectively. Samples from retail stores presented relatively low aflatoxin B incidences 14.5 ppb (1-57 ppb), but only 30% of the samples revealed aflatoxin level below 10 ppb. Laboratory segregated and carefully prepared butter from good grade nuts showed the lowest levels of this toxin (3.3 ppb; 2-6 ppb). The results showed that peanuts butter prepared by the street sellers and distributed by the retail stores are evidently hazardous to human health. There is therefore urgent need for strong form of quality control measures and public awareness. The use of excellent grade peanuts and care during processing and storage are priority. [ABSTRACT FROM AUTHOR]

Essential oils extracted from Citrus reticulata and Cymbopogon citratus were tested in vitro against the toxigenic strain of Aspergillus flavus, isolated from the tuberous roots of Asparagus racemosus, used in preparation of herbal drugs. The essential oils completely inhibited the growth of A. flavus at 750 ppm and also exhibited a broad fungitoxic spectrum against nine additional fungi isolated from the roots. Citrus reticulata and Cymbopogon citratus essential oils completely inhibited aflatoxin B1 production at 750 and 500 ppm, respectively. During in vivo investigation, the incidence of fungi and aflatoxin B1 production decreased considerably in essential oil-treated root samples. The findings thus indicate possible exploitation of the essential oils as effective inhibitor of aflatoxin B1 production and as post-harvest fungitoxicant of traditionally used plant origin for the control of storage fungi. These essential oils may be recommended as plant-based antifungals as well as aflatoxin B1 suppressors in post-harvest processing of herbal samples. [ABSTRACT FROM AUTHOR]

In agricultural areas, Aspergillus flavus, Aspergillus fumigatus and Aspergillus parasiticus are commonly identified in various feedstuffs and bioaerosols originated from feed handling. Some isolates belonging to these fungal species could produce mycotoxins and constitute a risk factor for human and animal health. In this study, Fourier-transform infrared spectroscopy was used for a rapid detection and characterization of 99 isolates collected from agricultural areas. The results showed a first cluster corresponding to strains previously attributed to the A. fumigatus group according to current taxonomic concepts, and a second cluster divided in 2 groups around reference strains of A. flavus and A. parasiticus species. The toxigenic capacity of isolates was evaluated by high performance liquid chromatography coupled to mass spectrometry. In the A. flavus group, only 6 strains of A. parasiticus and 4 strains of A. flavus were able to produce aflatoxins on culture media. FT-IR spectroscopy, respectively, allowed the differentiation of non-toxigenic and toxigenic A. flavus and A. parasiticus isolates at 75 and 100%. Discrimination between toxigenic and non-toxigenic A. fumigatus was not possible because all of the isolates produced at least one mycotoxin. [ABSTRACT FROM AUTHOR]

The ability of conidia of the human pathogenic fungus Aspergillus fumigatus to kill larvae of the insect Galleria mellonella was investigated. Conidia at different stages of the germination process displayed variations in their virulence as measured using the Galleria infection model. Non-germinating (‘resting’) conidia were avirulent except when an inoculation density of 1 × 107 conidia per insect was used. Conidia that had been induced to commence the germination process by pre-culturing in growth medium for 3 h were capable of killing larvae at densities of 1 × 106 and 1 × 107 per insect. An inoculation density of 1 × 105 conidia per insect remained avirulent. Conidia in the outgrowth phase of germination (characterised as the formation of a germ tube) were the most virulent and were capable of killing 100% of larvae after 5 or 24 h when 1 × 107 or 1 × 106 conidia, that had been allowed to germinate for 24 h, were used. Examination of the response of insect haemocytes to conidia at different stages of the germination process established that haemocytes could engulf non-germinating conidia and those in the early stages of the germination process but that conidia, which had reached the outgrowth stages of germination were not phagocytosed. The results presented here indicate that haemocytes of G. mellonella are capable of phagocytosing A. fumigatus conidia less than 3.0 μm in diameter but that conidia greater than this are too large to be engulfed. The virulence of A. fumigatus in G. mellonella larvae can be ascertained within 60–90 h if infection densities of 1 × 106 or 1 × 107 activated conidia (pre-incubated for 2–3 h) per insect are employed. [ABSTRACT FROM AUTHOR]

The use of nontoxigenic strains of Aspergillus flavus and A. parasiticus in biological control effectively reduces aflatoxin in peanuts when conidium-producing inoculum is applied to the soil surface. In this study, the movement of conidia in soil was examined following natural rainfall and controlled precipitation from a sprinkler irrigation system. Conidia of nontoxigenic A. flavus and A. parasiticus remained near the soil surface despite repeated rainfall and varying amounts of applied water from irrigation. In addition, rainfall washed the conidia along the peanut furrows for up to 100 meters downstream from the experimental plot boundary. The dispersal gradient was otherwise very steep upstream along the furrows and in directions perpendicular to the peanut rows. The retention of biocontrol conidia in the upper soil layers is likely important in reducing aflatoxin contamination of peanuts and aerial crops such as corn and cottonseed. [ABSTRACT FROM AUTHOR]

An investigation was undertaken to obtain data on the occurrence of aflatoxins and the aflatoxin producing potential of Aspergillus flavus strains isolated from dry fruit slices of quinces produced in jammu and Kashmir, India. A total of 147 A. flavus isolates recovered from dr fruit slices were grown in liquid rice flour medium and screened for the production of various aflatoxins by thin layer chromatography. The results showed that 23.14% of the tested isolates were aflatoxigenic, producing aflatoxins B1and B2 in varying amounts. Aflatoxins G1 and G2 were not detected. All 25 of the investigated market samples were also found to be aflatoxin B1 positive and the level of contamination ranged from 96 to 8164 μg/kg of the dry fruit which is quite high in comparison to the permissible level of 30 ppb. As per these results biochemical composition of dry fruit slices of quinces, along with climatic conditions seem to be very favourable for aflatoxin production by the toxigenic A. flavus strains. Therefore,monitoring of aflatoxins in dry fruit slices of quincesis recommended for this region. [ABSTRACT FROM AUTHOR]

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%) and F. proliferatum (1.8%). Eight species of Penicillium 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. [ABSTRACT FROM AUTHOR]

The luncheon meat samples analyzed, which were produced locally by the two main luncheon meat producing companies in Egypt were relatively highly contaminated either by moulds and yeasts in general, aflatoxigenic species and aflatoxin residues in particular. The most frequently encountered fungi from the samples were yeasts, Aspergillus niger, A. flavus, Penicillium chrysogenum, Rhizopus stolonifer, Mucor circinelloides. Less common were Cladosporium sphaerospermum, Alternaria alternata, Mycosphaerella tassiana, P. aurantiogriseum and P. oxalicum. The most important aflatoxigenic species, A. flavus, was isolated frequently. It was 10% of the total fungal isolates from both samples of the two companies. Seven luncheon meat samples out of 50 analyzed were positive for aflatoxin B1 or B1 and G1, while all samples were negative for aflatoxins B2, G2, M1 and M2. Aflatoxin B1 was detected only in 4 and 3 samples out of 25 analyzed from each of company A and B, respectively. The highest detectable level, 11.1 ppb, was recorded in a sample from company B and the least, 0.5 ppb, in a sample from company A. Aflatoxin G1, at concentration of 3.2 ppb, was detected in only one sample of the aflatoxin B1 – contaminated 3 samples of company B: this sample also had the highest level of aflatoxin B1. Some luncheon meat samples had higher numbers of aflatoxigenic A. flavus than others, however these samples were negative for aflatoxins. The hazardous potential of such contamination will be discussed. [ABSTRACT FROM AUTHOR]

Projects funded by International Development Research Centre (IDRC) of Canada and the European Commission have enabled the examination of more than 3000 samples of maize collected from all regions of Costa Rica at different stages, from the growing crop through storage to final sale, and at different water contents. Contamination with Aspergillus flavus was frequent and about 80% of samples contained more than 20 ng aflatoxins g-1 grain. Average contamination with aflatoxins in the Brunca Region was > 274 ng g -1 while that in other regions was < 70 ng g -1. Except in Brunca region, where it averaged 376 ng g -1, contamination of grain from commercial sources was slightly less than of that from farms (≤15 ng g-1). It appeared that samples kept on the cob after harvest contained almost no aflatoxin while shelled samples were frequently highly contaminated. Experiments were therefore done in Brunca and Huetar Atlantic Regions, utilising 34 experimental maize crops to study in detail the development of A. flavus and aflatoxin from before harvest, through postharvest treatment before drying and through storage for six months. A. flavus was isolated more frequently from maize shelled immediately after harvest than from that kept on the cob until it could be dried, and from more samples from the Brunca Region than from the Huetar Atlantic Region. Samples harvested with ≥18% water content often contained >70% of grains infected with A. flavus but sometimes there were few grains infected. As found in the initial survey, more aflatoxin contamination developed in shelled maize than in that handled on the cob during the period from harvesting to drying, especially if the delay was more than 5 days, and more in Brunca than in Huetar. Shelled grain contained 400–800 ng aflatoxin g -1 in Brunca but <100 ng g-1 in Huetar while grain kept on the cob contained <30 ng g-1, even with >18% water content. Incidence of Fusarium spp. exceeded 50% except where A. flavus colonized more than 80% of grains. [ABSTRACT FROM AUTHOR]

Culture filtrates of Trichoderma viride and Trichoderma harzianum were inhibitory of Fusarium moniliforme and, to a lesser extent, Aspergillus flavus. The degree of inhibition was, however, dependent on the carbon or nitrogen source incorporated into the medium. Scanning electron microscopy revealed the development of abnormal fruiting structures on exposure to some Trichoderma culture filtrate, while macroscopically, growth restriction and, in the case of A. flavus, altered colony colouration were observed. Based on the results of inverted colony culture, it would appear that some isolates of Trichoderma produce inhibitory volatile compounds. The production of possible antibiotics was also demonstrated. The aggressive behaviour (towards A. flavus and F. moniliforme) demonstrated by Trichoderma spp. may be partly explained by the liberation of extracellular enzymes by these fungi. An isolate of T. viride exhibited amylolytic, pectinolytic, proteolytic and cellulolytic activity. Based on the results of the present investigation, Trichoderma spp. are potential candidates for biocontrol of some mycotoxin-producing fungi, but there exists some doubt as to their osmotolerance within the air-dry seed. [ABSTRACT FROM AUTHOR]

The combined effects of water activity (a) and temperature on mycotoxin production by Penicilium commune (cyclopiazonic acid - CPA) and Aspergillus flavus (CPA and aflatoxins - AF) were studied on maize over a 14-day period using a statistical experimental design. Analysis of variance showed a highly significant interaction ( P ≤ 0.001) between these factors and mycotoxin production. The minimum a/temperature for CPA production (2264 ng g P. commune, 709 ng g A. flavus) was 0.90 a/30 °C while greatest production (7678 ng g P. commune, 1876 ng g A. flavus) was produced at 0.98 a/20 °C. Least AF (411 ng g) was produced at 0.90 a/20 °C and most (3096 ng g) at 0.98 a/30 °C. [ABSTRACT FROM AUTHOR]

Colonization of barley grain by Aspergillus flavus and formation of aflatoxin B in the presence of Penicillium verrucosum, Fusarium sporotrichioides, and Hyphopichia burtonii were studied over a three-week period in all combinations of 20 or 30 °C and 0.97, 0.95 or 0.90 a. Grain colonization was assessed initially by observing hyphal extension on the grain surface, using scanning electron microscopy, and then from the proportion of seeds infected and numbers of colony forming units (cfu) formed. Aflatoxin b concentrations were determined by enzyme linked immunosorbent assay using a monoclonal antibody. These studies showed that interaction between A. flavus and other fungi in paired culture had different effects on both colonization and aflatoxin formation depending on the species involved and environmental conditions. Germination of A. flavus spores was unaffected by the presence of other species on the grain surface. Subsequently, three principal patterns of A. flavus colonization of barley grain were observed through the incubation period in the presence of other fungal species: (a) colonization unaffected by the presence of other species; (b) colonization initially slower in the presence of other species but later differing little from pure cultures; and (c) colonization adversely affected by the presence of other species. Five main patterns of aflatoxin B production were observed relative to pure culture but with no consistent relationship with species, a, temperature or incubation period; (a) little changed; (b) increased slowly; (c) decreased; (d) enhanced; and (e, f) increased initially but later decreased to (e) the same level as in pure culture or (f) to less than in pure culture. Generally, production of aflatoxin B by A. flavus was less than in pure culture but sometimes was changed only slightly by the presence of P. verrucosum, F. sporotrichioides or H. burtonii or was temporarily enhanced. [ABSTRACT FROM AUTHOR]

A survey of the distribution of elastinolytic potential among 32 culture collection isolates of Aspergillus flavus, A. oryzae, A. parasiticus, A. sojae, A. nomius, and A. tamarii revealed this character to be highly conserved within Aspergillus Section Flavi. Furthermore, 144 isolates of A. flavus from environmental samples from six separate regions of the United States produced elastase on solid medium. Most previously described polymorphisms in elastinolytic potential were attributed to the toxicity of borate buffers. Replacement of borate with HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) resulted in detection of elastase production on solid medium by all tested fungal isolates except two that had been in culture over 50 years. In liquid culture, only isolates of A. flavus, A. tamarii, and A. oryzae accumulated elastase activity. Although isoelectric focusing revealed only one isoform (pI 9.0) of elastase in these culture filtrates, elastinolytic activity in filtrates was partially inhibited by both 1,10-phenanthrolene (2 mM) and phenylmethylsulfonylfluoride (2 mM), suggesting the presence of both metallo and serine elastinolytic proteinases. [ABSTRACT FROM AUTHOR]

A new method for growing Aspergllius flavus for experimental studies is presented. The system consists of a humidified vial with a thick septum pierced by a pin on which a glass fiber disc is affixed. The disc contains the test solution and inoculum plus medium. The method has been used to assess the effect of variations in culture conditions on production of aflatoxin B (AFB). The AFB level was affected by the amount of medium placed on the disc and type of disc material. The results for different types of glass fiber and quartz discs were compared with AFB produced by fungus grown in liquid medium or on paper discs. When compared to a liquid medium culture there was a 15 to 20-fold increase in AFB for one type of disc. Incubations with less than 14 µl of medium gave satisfactory results. A crude phosphatidylcholine preparation at a concentration of 0.7% of the medium resulted in a 4-fold increase in AFB. [ABSTRACT FROM AUTHOR]

Sixty nine samples of maize were collected from pre-harvest standing crops and on-farm storage facilities from 52 smallholder farms located within 4 regions of Honduras during October 1992 and November 1993. Samples were visually assessed for insect damage and fungal spoilage, and the mycoflora quantified on artificial media. The major components of the ear rot complex were: Fusarium moniliforme, F. moniliforme var. subglutinans, Penicillium species, Stenocarpella maydis, S. macrospora and Acremonium spp. Representative samples were also assayed for mycotoxin content. Fumonisin B was detected in all 24 samples tested at levels of between 68-6,555 (µg/kg), and aflatoxin was detected in 2 samples heavily contaminated with Aspergillus flavus. Moniliformin and tenuazonic acid were not detected in the samples tested. The implications of these findings for human and livestock health risk are discussed, together with possible strategies for controlling these pathogens. [ABSTRACT FROM AUTHOR]

During an investigation of the mycoflora on oilseed rape, the predominant fungal species present in 20 samples collected from Catalonia (Spain) were Alternaria alternata (Fries) Keissler, Penicillium spp. and Aspergillus flavus. None of the 20 samples analyzed presented contamination by Alternaria mycotoxins (tenuazonic acid, alternariol, alternariol methyl ether, altertoxin I and altertoxin II). Only aflatoxin B was detected in 1 of the 20 samples analyzed, with a concentration of 0.25 ppb. Of the 40 Aspergillus flavus strains isolated from oilseed rape samples, only 3 revealed aflatoxigenic capacity. None of the Penicillium spp. isolated from oilseed rape samples revealed mycotoxigenic capacity (citreoviridin, griseofulvin, citrinin, patulin and penicillic acid). [ABSTRACT FROM AUTHOR]

Figs in an orchard were inoculated with an aflatoxigenic Aspergillus flavus strain in two ways by spore injection or by dusting at three maturation stages: firm ripe, shrivelled, and dried. Fruits were individually examined for fungal development and analyzed for aflatoxin B (AF B) after 2, 4, 6, 8 and 10 days. Fruit injected at the first stage showed fungal development and AF B contamination within two days. The toxin level increased sharply to 1 ppm after 10 days. The mean level of AF B (284.75 ng/g) was significantly higher than those observed in other conditions. Figs dusted at the first stage showed only a tiny fungal growth even after 10 days. AF B appeared after 6 days with a low frequency (35%), mean level (7.6 ng/g) and a great variation among figs (0.22-15 ng/g). Among fruits inoculated during the shrivelled fig and dried fruit stages, no fungal growth was observed and AF B was detected with a lower incidence in association with low mean levels (less than 1.25 ng/g). Methods of prevention of aflatoxin contamination at the critical step, the firm ripe stage, are discussed. [ABSTRACT FROM AUTHOR]

Environmental control plots adjusted to late season drought and elevated soil temperatures where inoculated at peanut planting with low and high levels of conidia, sclerotia, and mycelium from a brown conidial mutant of Aspergillus parasiticus. Percentage infection of peanut seeds from undamaged pods was greatest for the subplot containing the high sclerotial inoculum (15/cm soil surface). Sclerotia did not germinate sporogenically and may have invaded seeds through mycelium. In contrast, the mycelial inoculum (colonized peanut seed particles) released large numbers of conidia into soil. Soil conidial populations of brown A. parasiticus from treatments with conidia and mycelium were positively correlated with the incidence of seed infection in undamaged pods. The ratio of A. flavus to wild-type A. parasiticus in soil shifted from 7:3 to 1:1 in the uninoculated subplot after instigation of drought, whereas in all subplots treated with brown A. parasiticus, the ratio of the two species became approximately 8:2. Despite high levels of brown A. parasiticus populations in soil, native A. flavus often dominated peanut seeds, suggesting that it is a more aggressive species. Sclerotia of wild-type A. parasiticus formed infrequently on preharvest peanut seeds from insect-damaged pods. [ABSTRACT FROM AUTHOR]

Thirty-two isolates of Aspergillus flavus were obtained from various sources in Hungary. All isolates were morphologically identified as A. flavus and three atypical variants were confirmed as A. flavus by comparing their DNA with an ex type culture of A. flavus. None of these isolates produced aflatoxins when tested on coconut agar or grown on rice medium and culture extracts examined by thin layer chromatography. Also, none of the isolates converted sterigmatocystin, O-methyl sterigmatocystin, norsolorinic acid, or sodium acetate to aflatoxin. However, 59% of the isolates produced cyclopiazonic acid based on thin layer chromatographic analysis of culture extracts. The isolates that lack the ability to produce both aflatoxin and cyclopiazonic acid are potential candidates for use in bicontrol studies. [ABSTRACT FROM AUTHOR]

Two hundred strains of the Aspergillus flavus group isolated from groundnuts (peanuts) growing in Israel were examined for their ability to produce mycotoxins in potato dextrose (PD) broth. Almost 77% of the isolates produced aflatoxin; aflatoxins B and B were formed by most of the isolates. Simultaneous production of aflatoxins of groups B and G was detected in only 0.5% of the isolates. Microscopic examination revealed that 98% of the isolates were A. flavus and only 2% A. parasiticus. Cyclopiazonic acid (CPA) was detected in 22.5% of the isolates, including 3.5% that produced only CPA. Sterigmatocystin was detected in only 2% of the isolates and only one isolate produced aflatoxin simultaneously with CPA and sterigmatocysin. The dry weight (DW) of mycelium, 7 days after inoculating the medium, was between 71-110 mg/30 ml medium in more than 70% of the isolates. A general decrease in the pH was observed and 75% of the isolates reduced the pH to 4.5 or below. After 14 days, a small increase in DW and an increase in the pH toward neutrality was observed. On PD agar, 30% of the isolates produced sclerotia, including 5% that produced them profusely. No correlation between mycelial growth, changes in pH of the medium, sclerotium formation, and aflatoxin accumulation could be observed. The mycotoxigenic potential of the A. flavus strains isolated from groundnuts seems to be relatively high and may present a potential threat to human and animal health. [ABSTRACT FROM AUTHOR]

In Kanagawa Prefecture, located in central Japan, aflatoxin-producing Aspergillus flavus was isolated in 4 (2.5%) of 160 field soil samples. In the 4 fields, whose soil contained aflatoxin-producing A. flavus, the annual average temperature of the sampling sites of the soil ranged from 13.8 to 15.1°C. Of all the isolated strains of aflatoxin-producing A. flavus, 4 strains, isolated from a single soil sample, produced large amounts of aflatoxin B and B when incubated in coconut agar, peanut agar, peanuts or trilaurin-added rice, although they did not produce aflatoxin when incubated in rice, yeast extract-sucrose broth or sucrose-low salts broth. [ABSTRACT FROM AUTHOR]

Pectinase and sclerotium production by strains of Aspergillus flavus were determined with a pectinase culture plate assay and a Cz 3% NaNO medium plate assay. In the A. flavus population, 51% of isolates produced sclerotia, the toxigenic strains showing a tendency to have smaller sclerotia. Strains producing both abundant small sclerotia and a large quantity of aflatoxin were not found. There was no linear correlation between the amount of aflatoxin produced and the number of sclerotia. Levels of pectinase produced by the toxigenic strains were higher than that of the non-toxigenic strains, and this character was more obvious in the sclerotium-producing strains than in the non-sclerotium-prodcing strains. In the A. flavus population from Zhejiang in which the toxigenic strain rate was low, toxigenic strains may require higher levels of pectinase to compete with the non-toxigenic strains when infecting foodstuffs. [ABSTRACT FROM AUTHOR]

The fungal flora of 6 Asian medicinal plants, Aerva lanata (Linn.) Juss. Alyssicarpus vaginalis D.C., Tribulus terrestris Linn. Adhatoda vasica Nees., Centella asciatica (L.) Urb., Cardiospermum halicacabum Linn. was determined. After surface disinfection Aspergillus spp. were most frequently observed. Aspergillus flavus, isolated from Alyssicarpus vaginalis and Aerva lanata produced aflatoxins in culture. Aflatoxin B was also detected in a sample of Aerra lanata at a level of 0.5 μg/g. Plant material destined for medicinal use should be stored carefully prior to its use to prevent growth of naturally occurring toxigenic mold fungi. [ABSTRACT FROM AUTHOR]

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 °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 flourescence 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). [ABSTRACT FROM AUTHOR]

A survey was carried out in Reunion island to obtain data on the occurrence of fungi, aflatoxigenic strains of Aspergillus flavus, aflatoxins, total aerobic bacteria and salmonellae of 150 samples of mixed poultry feeds and raw materials. These were collected at five farms over a 3-month period during the warm rainy season. White corn and Brazilian soybean meal seemed to present a better microbiological quality than yellow corn and US soybean meal. Mixed poultry feeds presented a high total mold count reflecting the mold flora of raw materials. The most frequent and abundant fungi were Aspergillus flavus, A. glaucus group, Fusarium spp., Penicillium spp., A. candidus, Mucor spp., A. restrictus, Scopulariopsis spp., Cladosporium spp. and A. versicolor. Of the 118 A. flavus strains screened, 42 (35.6%) were aflatoxigenic. Yellow corn samples were the most frequently contamined with aflatoxigenic strains (54.5%), followed by mixed feeds (44%). Of the 66 samples tested, 24 (36%) contained aflatoxins (traces to 22 ng/g). A good correlation seemed to exist between presence of at least one aflatoxigenic strain per sample and presence of aflatoxins. [ABSTRACT FROM AUTHOR]

Thirty four samples of rice bran, of which 9 were from raw (untreated) rice (RR) and 25 from parboiled rice (PbR) were collected from commercial rice mills in and around Madras and analysed for storage mycoflora and mycotoxins. Fungi of the Aspergillus flavus group were present in 29 of the 34 samples (8 from RR and 21 from PbR) in quantities ranging from <1-432 thousand propagules/g, though not always as the dominant mycoflora. Fungal numbers were usually higher in RR than in PbR samples. Five of the 9 RR samples and 6 of the 25 PbR samples were positive for aflatoxins. Among 29 isolates of A. flavus obtained from the bran samples, 16 isolates −6 from RR bran and 10 from PbR bran - were found to be toxigenic in vitro. Some isolates of A. candidus also seemed to produce aflatoxin and other fluorescent substances. [ABSTRACT FROM AUTHOR]

Cottonseeds having fluorescent fibers were harvested from fields in Arizona and examined utilizing light microscopy and transmission electron microscopy. The occurrence of fluorescent fibers indicated that seeds had been infected by Aspergillus flavus during development. Presence of A. flavus was verified by plating portions of seeds with fluorescent fibers. Hyphae, conidial heads, and conidia were identified readily in differentially-stained cotyledon tissue processed for light microscopy. Utilization of transmission electron microscopy permitted observations on lignified seed coats and cotyledons of mature cottonseeds. Hyphae were located throughout the cotyledon and in the nonlignified layers of the seed coat. The identification of hyphae in cross sections of vessel elements within the seed coat provided ultrastructural evidence supporting the hypothesis that A. flavus may enter seeds via the vascular tissue. Controls for the microscopy studies included observations on cottonseeds with no visual signs of infection and on laboratory-grown cultures of A. flavus. These observations demonstrated that the hyphae localized within fluorescent seeds had features characteristic of A. flavus and that fungal-like structures do not occur within uninfected seeds. [ABSTRACT FROM AUTHOR]

Non-aflatoxin-producing isolates of Aspergillus flavus from nature and isolates of A. flavus that had lost their toxigenic trait following laboratory transfer were compared biochemically. After the addition of aflatoxin B precursors sterigmatocystin or O-methylsterigmatocystin to whole cell cultures, the non-toxin producing isolates from nature remained non-toxigenic while toxigenicity was restored in the nontoxigenic laboratory strains. Results imply a lack of enzymes needed for biochemical conversions of precursors to aflatoxin B in natural non-producers and suppression of these enzymes in the nonproducing laboratory strains. [ABSTRACT FROM AUTHOR]

In recent years Aspergillus flavus and aflatoxin production have been noted on several occasions in grain preserved with formic acid. Samples of mouldy barley treated with formic acid and stored in an open bin were investigated for the presence of fungi. In the lower part of the bin there was a clear dominance of Fusarium sporotrichioides, and deoxynivalenol and neosolaniol were detected. A. flavus and A. fumigatus were also present. Paecilomyces variotii occurred, almost as a pure culture, in the upper part of the bin, but no patulin was found. Cultivation of four fungal isolates from these genera on laboratory substrates containing formic acid showed P. variotii to be the most tolerant to formic acid, withstanding 150 mM, but still without patulin production. F. sporotrichioides and A. fumigatus tolerated only 6 mM formic acid. The growth of A. flavus was reduced and atypical at 60 mM formic acid. Pretreatment of A. flavus spores with formic acid increased aflatoxin production about 800 times. [ABSTRACT FROM AUTHOR]

The effect of different nitrogen sources and varying glucose concentration on aflatoxin production by a toxigenic and non-toxigenic strain of Aspergillus flavus was studied. Greatest production (3.8 ppm) of aflatoxin B was produced in a synthetic medium when casamino acids were supplied as the nitrogen source. Optimum sugar concentration for aflatoxin B production ranged between 3 and 10 g/100 ml. There was no appreciable difference in the metabolic behaviour between toxigenic and non-toxigenic strains of A. flavus when dry mycelial weight, total proteins, non-protein nitrogen and reducing sugar were the criteria. [ABSTRACT FROM AUTHOR]

Twenty-seven mature cotton bolls with Aspergillus flavus Link colonies naturally occurring on the surface of the boll or lint were collected in the field in Arizona along with their subtending stems and peduncles. Bolls inoculated through the carpel wall 30 days after anthesis were allowed to mature in the field and were collected in the same manner. The seed and stem and peduncle sections of each boll were surface-sterilized, plated on agar media and observed for A. flavus. Seventy-eight percent of the naturally contaminated bolls with A. flavus in the seed also had the fungus in the stem and peduncle, whereas only 31% of the naturally contaminated bolls with no A. flavus in the seed had the fungus in the stem or peduncle. This difference was significant (P=0.0125), indicating a positive relationship between seed infection and stem and peduncle infection. All of the bolls inoculated through the carpel wall had A. flavus in the seed, but only 11% of the stem and peduncle sections were infected, indicating that the fungus does not readily grow downward from the boll into the supporting stem or peduncle. This unidirectional pattern of movement (upward) was further substantiated in greenhouse experiments where cotton seedlings were inoculated at the cotyledonary leaf scar with A. flavus and plants were sequentially harvested, surface sterilized and plated. Aspergillus flavus was isolated from the cotyledonary leaf scar, flower buds, developing bolls, and stem sections in the upper portion of the plant. It was never isolated from roots or stem sections below the cotyledonary node, again indicating that the fungus does not readily move downward through the plant. [ABSTRACT FROM AUTHOR]

A total of 126 fungal strains belonging to the Aspergillus flavus group isolated from commercial poultry mixed feeds were studied. One hundred and twenty-five were identified as A. flavus and one as A. parasiticus. Forty nine strains (39%) produced aflatoxins on a crushed moist wheat medium (28 °C/10 days), whereas only sixteen (13%) showed specific fluorescence on Aflatoxin-Producing Ability Medium. In both media, mainly aflatoxins B and B were detected, the average concentration of aflatoxins being 4294+/−1083 μg/kg in crushed moist wheat medium, and 877+/−257 μg/kg in Aflatoxin-Producing Ability Medium. [ABSTRACT FROM AUTHOR]