Further studies on Egyptian soil fungi: Succession of sugar and osmophilic fungi in soil amended with five organic substrates

The sugar and osmophilic fungal composition of soils amended with five organic substrates (newspaper, orange peel, bromegrass leaves, wheat straw and wood sawdust) was estimated after 2, 4, 6, 8 and 10 weeks using the dilution plate method on glucose and 50% sucrose Czapek's agar media. Wheat straw was the best substrate for total counts of both sugar and osmophilic fungi followed by newspaper, bromegrass leaves, wood sawdust and orange peel. Wood sawdust supported the highest average counts of total sugar fungi, Fusarium, Mucor, Scopulariopsis, Trichoderma and Trimmatostroma spp.; Newspaper, of Aspergillus (8 spp.), Penicillium (4 spp.) and Chaetomium sp. bromegrass leaves of Cladosporium sp., Humicola sp. and Sporotrichum sp.; orange peel, ofAlternaria sp., Circinella sp. and Stachybotrys sp.; and wheat straw, of Botryotrichum sp. and Myrothecium sp. Bromegrass leaves and orange peel supported the highest average counts of total osmophilic fungi, Aspergillus (10 spp.), Cladosporium sp. Paecillomyces sp. and Rhizopus sp.; and of Stemphylium sp., Trichoderma sp., Humicola sp. and Circinella sp. respectively; wheat straw, of Epicoccum sp., Scopulariopsis sp. and Trichothecium sp.; newspaper, of Penicillium (4 spp.) and Alternaria sp.; and wood sawdust of Curvularia sp. and Fusarium (3 spp.). The best colonizers throughout the experimental periods were Aspergilus and Penicillium spp.     

Aflatoxin and aflatoxicosis

A wide range of fungi, amounting to fifty-six species belonging to twenty-two genera, have been recovered from animal feeds and foodstuffs.The most frequent fungi were Aspergillus niger, and A. flavus, followed by Mucor racemosus, Alternaria alternata, Rhizopus stolonifer, Penicillium corylophilum and P. notatum. Three genera were found to be of moderate occurrence, namely, Mucor, Rhizopus and Alternaria. The three following genera were of low occurrence: Cladosporium, Fusarium, and Neurospora.The fluorescence method of detecting aflatoxin-producing strains demonstrated that only one isolate of A. flavus possesses this property. Certain species of Penicillium and Aspergillus produced fluorescent substances (metabolites) similar in color to B and G aflatoxin. These substances were subsequently proved not to be aflatoxin by (TLC) chromatography.The animal and public health significance from such toxins was also discussed.     

Ecology of fungi associated with moist stored barley grain

A study of the ecology of fungi associated with moist stored barley grain was carried out on samples from a partially sealed silo in Cumberland. Samples were collected during harvest and after storage at high moisture content in a partially sealed silo. At harvest, the field fungi Alternaria, Fusarium, Cladosporium and Mucor spp. were dominant both in the husks and in the dehusked, surface-sterilized grain but they were rapidly succeeded by the storage fungi: Aspergillus, Penicillium and Absidia spp. then constituted about 80 % of the initial invasion. At this stage internal grain infection rarely exceeded 5 % compared to 65–80 % infection in the husks. In subsequent months, there was a remarkable increase in invasion, mainly by Penicillium, Aspergillus and Absidia spp. and by yeasts, which appeared both in the husks and in dehusked grain. In the third month of storage, Penicillium cyclopium, P. roqueforti, Absidia corymbifera, Aspergillus candidus, A. terreus and yeasts became dominant. The thermophilous species Aspergillus fumigatus, A. terreus, Absidia corymbifera, Mucorpusillus and Dactylomyces crustaceus were isolated from husks and in dehusked grain incubated at 37 and 45 °C. Most grains were infected significantly by single genera.     

Separation of Fungal Propagules by Partition in Aqueous Polymer Two-Phase Systems

Conidia of Penicillium chrysogenum and Penicillium frequentans and sporangiospores of Rhizopus rhizopodiformis, Rhizomucor pusillus, and Mucor racemosus were subjected to partition in aqueous polymer two-phase systems. The partition behavior differed drastically between the conidia of the two Penicillium species and the sporangiospores of the three species of phycomycetes. This difference in partition behavior can be used for purification of fungi belonging to different taxonomical groups. P. frequentans was completely separated from M. racemosus by two extractions, whereas four extractions were needed to purify M. racemosus. This method was used on an air sample from a locality where wood fuel chips are handled. The conidia of the fungi Trichoderma viride and Rhizopus rhizopodiformis were removed completely by only two extractions.     

Detection of 3-nitropropionic acid and cytotoxicity inMucor circinelloides

Mucorales are regarded as the aetiological agents of Mucormycosis. Their capabilities to produce mycotoxins are not profoundly investigated, in contrast to those of the fungi from the generaPenicillium, Aspergillus, orFusarium. The aim of this study was to isolate and identify fungi of the order Mucorales and investigate mycotoxins production. Twelve samples of visibly moulded grass silage and eight samples of damaged whole crop maize silage were analysed. Malt extract agar plates were used for sub cultivation. Three fungal species of the order Mucorales were isolated from grass silage, which were identified by their macro-and micro-morphology asAbsidia corymbifera, Mucor circinelloides andRhizopus stolonifer. The cytotoxicity ofMucor circinelloides extract was analysed using the cytotoxicity test (MTT assay) and the result, showed a low cytotoxicity. Additionally extracts fromAbsidia corymbifera, Mucor circinelloides andRhizopus stolonifer were tested for mycotoxin-production using an LC/MS/MS-based multimycotoxin method. 3-nitropropionic acid was detected in the culture extract ofMucor circinelloides. Presented at the 30^th Mykotoxin Workshop Utrecht, Netherlands, April 28–30, 2008.     

Low occurrence of patulin‐ and citrinin‐producing species isolated from grapes

Aims: To assess the ability of fungi isolated from grapes to produce patulin and citrinin. Methods and Results: A total of 446 Aspergillus isolates belonging to 20 species and 101 Penicillium isolates were inoculated in Czapek yeast extract agar and yeast extract sucrose agar and incubated for 7 days at 25°C. Extracts were analysed for patulin and citrinin by thin‐layer chromatography. None of the isolates of Aspergillus spp. produced either patulin or citrinin. Patulin was produced by three isolates of Penicillium expansum and two of Penicillium griseofulvum. Citrinin was produced by five isolates of P. expansum, two of Penicillium citrinum and one of Penicillium verrucosum. Conclusions: Our results show that the Aspergillus and Penicillium species commonly isolated from grapes are not a source of the mycotoxins, patulin and citrinin. Significance and Impact of the Study: The possibility of co‐occurrence of patulin and citrinin with ochratoxin A in grapes and grape products remain low, owing to the low frequency of isolation of potentially producing species.     

Evaluation of five sugar beet varieties for root-knot nematode and root-rot fungal infection

Survey results during 2010–2011 season revealed that the Meloidogyne spp., Helico–tylenchus spp. and Tylenchorhynchus spp. were the common plant parasitic nematodes in the rhizospheres of sugar beet in El-Behera, El-Fayoum and Beni Sueif Governorates. Aspergillus spp., Aspergillus niger, Fusarium spp., Penicillium spp., Penicillium chrysogenum, Penicillium citrinum, Rhizoctonia spp., Rhizopus nigricans, Trichoderma spp. and others were also the common fungi in the same rhizospheres. The rhizosphere of El-Behera Governorate was highly infected with Meloidogyne spp., Fusarium spp. and Rhizoctonia spp., compared to the rhizospheres of El-Fayoum and Beni Sueif, respectively. Evaluation of five of sugar beet cultivars, viz. Chems, Dema-Poly, DS 9001, Manila and Ras-Poly to infection, with Meloidogyne incognita (root-knot nematode), Fusarium solani and Rhizoctonia solani (root rot disease) was carried out under naturally field infection condition in the Nubariya region, Behera Governorate during 2011–2012 season. Host susceptibility rating revealed that the varieties of Ras-Poly, DS 9001 and Manila can be considered as susceptible, while the varieties of Dema-Poly and Chems can be considered as highly susceptible. Pathological results showed that the varieties of Dema-Poly and Ras-Poly were highly infected with F. solani, while not infected with R. solani. The varieties of DS 9001, Manila and Chems were moderately infected with the two pathogens.     

Mycoflora of bean,broad bean,lentil, lupine and pea seeds in Saudi Arabia

Using the seed-plate method on glucose-Czapek's agar at 28 °C (±2 °C), 24 genera and 60 species in addition to two varieties were collected from bean, broad bean, lentil, lupine and pea seeds. The most frequent genera were Aspergillus (16 species + two varieties), Penicillium (14 species), Rhizopus (1 species) and Yeasts, followed by Fusarium (3 species), Mucor (4 species) and Drechslera (3 species). From the preceding genera, Aspergillus niger, A.flavus, Penicillium citrinum, Rhizopus stolonifer, Fusarium moniliforme or F. oxysporum, Mucor hiemalis or M. racemosus and Drechslera spicifera were the most prevalent species.     

The water content/water activity relationship of cured tobacco and water relations of associated spoilage fungi

The relationship between moisture content and water activity (a_w) in cured tobacco was significantly influenced by sugar content. Overall, high sugar tobaccos such as Oriental and Virginia had a higher moisture content at any given water activity compared to low sugar tobaccos such as Burley. Virginia and Burley were both predominantly colonised by Aspergillus and Penicillium spp. Of these, about 80% of isolates could germinate at between 0·75 and 0·85 a_w, equivalent to moisture contents of between 18% and 24% in Burley and between 22% and 31% in Virginia. Growth of the dominant Aspergillus and Penicillium spp. was much slower on Virginia and Burley tobacco extract than on malt extract agars over the range 0.85 to 0.98 a_w. For some species the optimum a_w for growth on tobacco extract medium was altered from that on the richer malt extract agar and for some there was also a significant difference in growth between Virginia and Burley extract agars. The mould-free storage periods for five different tobacco types was influenced by a_w. Visible moulding occurred within 7–14 days at 0.85–0.90 a_w but only after about six months at 0.70–0.75 a_w. There were some differences in rate of moulding between tobacco types as well as in the range of fungi isolated at different a_w storage levels.     

Mycoflora of mangrove mud

Summary Thirty-nine species and four varieties of fungi are reported from mangrove mud collected from Kagh Island, Port Canning and Diamond Harbour, West Bengal, India. Of these forms twelve species and two varieties belonged to genusAspergillus and three species belonged, each toMucor, Penicillium andPreussia. Two species both ofFusarium andTrichoderma were also isolated. Two new species,Cladosporium indicum andPaecilomyces indicus and five other interesting fungi are described in detail.     

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.     

A survey of outdoor and indoor airborne fungal spora in the Redemption City, Ogun State, south-western Nigeria

The concentration and biodiversity of airborne fungi of the Redemption City, an immense campground for Christian faithful and the temporary site of the Redeemer’s University in south-western Nigeria, was studied between February and May 2011 using the culture plate method. The study was undertaken to assess the concentrations of fungal spores and their health implication in this ever-busy environment. Fifteen different sites classified as closed or open were selected. During the experiment, a total of 228 colonies were counted, and 29 fungal species belonging to 26 genera were isolated which include the following: Aspergillus flavus, Aspergillus niger, Bipolaris spp., Chrysosporium spp., Cladosporium spp., Coniothyrium corda, Curvularia spp., Diplodia spp., Fusarium spp., Gliocladium spp., Monilia spp., Mucor spp., Mucor plumbeus, Penicillium spp., Phycomyces spp., Phytophthora spp., Pilobolus spp., Pyrenochaeta spp., Rhizopus stolonifer, Torula spp., Trichoderma spp. and Trichophyton spp. The most frequently occurring fungi were A. niger, C. corda and M. plumbeus, while the least recorded were Torula and Trichophyton species. Majority of the fungi isolated are known allergens; they could also be opportunistic causing various diseases in man. There is therefore a dire need for good sanitation practices within the studied areas of the camp.     

Mycoflora isolation and molecular characterization of Aspergillus and Fusarium species in Tunisian cereals

Wheat, barley and maize are the mainly consumed cereals in Tunisia. This study aimed to determine the mycoflora of these cereals with special focus on the mycotoxigenic Aspergillus and Fusarium species. Freshly harvested samples and other stored samples of each type of cereal (31 and 34 samples, respectively) were collected in Tunisia and cultured for fungal isolation and identification. Identification of fungal genera was based on morphological features. Aspergillus and Fusarium species were identified by species specific PCR assays complemented with DNA sequencing. Alternaria (70.83%), Eurotium (62.50%), Aspergillus (54.17%) and Penicillium (41.67%) were the most frequent fungi isolated from wheat. Penicillium (75%), Aspergillus (70%), Eurotium (65%) and Alternaria (65%) were the most frequently recovered genera from barley. The predominant genera in maize were Aspergillus (76.19%), Eurotium (42.86%), and Penicillium (38.09%). Aspergilllus, Penicillium, Fusarium and Alternaria were detected in both stored and freshly harvested grain samples. The frequencies of contamination with Aspergillus, Fusarium and Alternaria were higher in freshly harvested samples, whereas Penicillium species were more frequent in stored samples. The predominant Aspergillus species detected were A. flavus and A. niger. The Fusarium species detected were F. equiseti, F. verticillioides, F. nygamai, and F. oxysporum. This study suggested the potential risk for Aflatoxins and, to a lesser extent, for Ochratoxin A in Tunisian cereals. This is the first survey about mycoflora associated with wheat, barley and maize in Tunisia.     

Imaging techniques for the detection of stored product pests

Stored grains are subject to deterioration and losses through various factors, but mainly insects and fungi. Various techniques are employed to detect stored product pests; however, there is an urgent need for an industrial-scale on-line detection technique. Near-infrared hyperspectroscopic imaging and soft X-rays have shown the potential for real-time application. These techniques are particularly effective for detecting internal infestations of stored grains. The digital images of the scanned objects are analyzed for various spectral and image features using statistical techniques such as complex multivariate tools. Classification accuracies as high as 80–100 % have been achieved for various pest and grain combinations. Dual-energy X-rays have been shown to detect the concealed eggs of stored product insect pests. The main threats to stored cereals come from Aspergillus spp., Penicillium spp., and Fusarium spp., which may produce mycotoxins. These imaging techniques have shown good results in the detection of fungal infections of stored grain.     

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.     

Invasion of peanut fruits by Aspergillus flavus and other fungi

Intensive studies of the mycoflora of Dixie Spanish peanuts grown under different combinations of tillage, organic matter, and previous crop revealed no consistent reduction in either Total Fungi,A spergillus spp.A. niger, A. flavus, Penicillium spp., orFusarium spp. for any of the 16 combinations studied. The level of fungous invasion in apparently sound peanut shells and seeds was consistently high, but the species composition of this flora changed as the season progressed.Penicillium was the most common genus, being isolated from over half the shells sampled. The level in seeds was lower, but in both shells and seeds there were morePenicillium spp. in fruits in the ground than in fruits from stacks.A spergillus spp. occurred at lower levels thanPenicillium, but they also decreased in fruits in the stack. Of particular interest is the fact that at all three collecting dates the amount ofAspergillus isolated from seeds was at least twice that from shells. The lower frequency of certain fungi in fruits from stacks is undoubtedly related to the lower moisture level in the drying fruits. Other fungi, especiallyFusarium spp., increase sharply in the dried tissues so that the overall level of invasion remains high. This may be of considerable practical importance since certain species ofFusarium are now known to produce toxins. Whether the increase in these fungi is due to their ability to grow under conditions of low moisture or is related to altered interspecific competition resulting from differences in the abilities of other species to survive the dry habitat, or both, is uncertain.

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Zusammenfassung Eingehende Untersuchungen der Mykoflora der Dixie Spanish Erdnüsse unter verschiedenen Kombinationen von Ackern, organischem Material und vorherigem Anbau zeigten keine beständige Verminderung des gesamten Pilzgutes,Aspergillus spp.A. niger, A. flavus, Penicillium spp., oderFusarium spp., für keine der 16 Kombinationen. Der Grad der Pilzinvasion in anscheinend gesunden Erdnußhülsen und Samen war ständig hoch, aber die Artzusammensetzung dieser Flora wechselte mit der fortschreitenden Jahreszeit.Penicillium war die häufigste Gattung und war von der Hälfte der Hülsen isoliert. Der Grad in den Samen war niedriger, aber sowohl in Hülsen als auch in Samen war mehrPenicillium spp. in Früchten im Boden als in Früchten im Stapel.Aspergillus spp. kam zu einem niedrigeren Grade alsPenicillium vor, aber sie nahmen auch in Früchten im Stapel ab. Von besonderem Interesse ist die Tatsache, daß die Anzahl vonAspergillus spp., die von Samen isoliert worden sind, war mindestens das Zweifache von denen, die von Hülsen erhalten worden sind, an allen drei Sammeldaten. Die niedrigere Häufigkeit gewisser Pilze in Früchten vom Stapel ist zweifellos mit dem niedrigeren Feuchtigkeitsgrad der trockenen Früchte im Zusammenhang. Andere Pilze, insbesondereFusarium spp., nahmen im trockenen Gewebe stark zu, so daß der allgemeine Grad der Invasion hoch blieb. Dies mag von einer besonderen, praktischen Gewichtigkeit sein, da gewisse Arten vonFusarium bekannterweise Toxine produzieren. Ob die Zunahme dieser Pilze die Folge ihrer Fähigkeit ist, unter Bedingungen niedriger Feuchtigkeit zu wachsen oder sie mit dem veränderten interspezifischen Wettbewerb im Zusammenhang steht als Ergebnis der Unterschiede in der Fähigkeit anderer Pilze, in trockener Umgebung zu überleben oder beide, ist ungewiß.

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Journal Series Paper No. 612, University of Georgia College of Agriculture Experiment Stations, College Station, Athens. This research was supported in part by funds from Cooperative Agreement No. 12-14-100-9378(34), administered by Peanut Investigations, Oilseed and Industrial Crops Branch CRD, ARS, USDA. Accepted for publication: 20. IX. 1969     

The fungal flora of soft fruits in relation to storage and spoilage

The fungal population of freshly harvested strawberries, raspberries and blackberries was studied throughout the fruiting seasons and the relative importance of fungi responsible for spoilage of stored fruit from different harvest dates was assessed. Botrytis cinerea, Cladosporium spp., Penicillium spp., Aureobasidium pullulans and Mucor spp. were consistently present on all three fruits while Colletotrichum gloeosporioides and Didymella applanata were prevalent on raspberries only and Epicoccum purpurascens and Alternaria spp. on blackberries only. Botrytis cinerea and Mucor mucedo were the predominant spoilage organisms of all three fruits, although the relative importance of the two fungi changed as the season progressed, especially for strawberries. Rhizopus stolonifer, Cladosporium spp., Penicillium spp. and Colletotrichum gloeosporioides were only of minor importance as spoilage organisms.     

Hydrolysis of nitriles and amides by filamentous fungi

Screening of culture collection afforded nitrile-utilizing fungi belonging to genera Aspergillus, Talaromyces and Penicillium. Fusarium solani O1 was enriched from soil using 3-cyanopyridine as the sole source of nitrogen. This strain, and Penicillium multicolor CCF 2244 (the best one of the culture collection strains), showed comparable specific benzonitrile-hydrolyzing activities (0.95 and 0.87 μmol of benzoic acid h⁻¹ mg⁻¹ of dry cell weight at 28 °C, respectively). These fungi showed similar substrate specificities for substituted benzonitriles and heterocyclic nitriles but different pH and temperature optima (pH 8 and 38 °C for P. multicolor, pH 7 and 48 °C for F. solani). Amides as by-products were produced from some heterocyclic nitriles. Both fungi showed an amidase activity for nicotinamide.     

Evaluation of Etest Performed in Mueller–Hinton Agar Supplemented with Glucose for Antifungal Susceptibility Testing of Clinical Isolates of Filamentous Fungi

Although reference broth microdilution protocol is currently available for filamentous fungi antifungal susceptibility testing (AFST), simpler alternatives as Etest^® tend to be favoured in clinical routine, making their validation of utmost importance. In this study, Etest^® method using 2 % glucose supplemented Muller–Hinton agar was compared to the Clinical and Laboratory Standards Institute (CLSI) M38-A2 protocol for filamentous fungi AFST. The echinocandins, caspofungin and anidulafungin, the azoles voriconazole and posaconazole, and the polyene amphotericin B were tested against 48 Aspergillus spp., seven Fusarium spp., one Beauveria bassiana and three Paecilomyces lilacinus isolates. The majority of the isolates were susceptible to the antifungals tested, and the overall level of agreement between the CLSI and Etest methods was 71.9 % for one dilution and 99.7 % when using two dilutions. Since interpretative breakpoints for filamentous fungi employing the CLSI or Etest methods are not available yet, the established epidemiological cut-off values for Aspergillus spp. were used to distinguish wild-type isolates from those with acquired resistance mechanisms. Forty-five Aspergillus strains did not evidence resistance mutations.     

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.