Occurrence of itraconazole-tolerant micromycetes in the soil and food products

Unexpected pathogens from the environment represent considerable risk for humans with impaired health. We examined the occurrence of itraconazole tolerant micromycetes in soil and in maize products. Five concentrations of itraconazole (2.5–12.5 μg/mL) selected according to known treatment schedules for human patients were incorporated into Sabouraud agar with chloramphenicol and Rose Bengal and diluted samples were inoculated onto the agar surface. After 7-d growth at 22°C colonies ofAlternaria sp.,Aspergillus clavatus. A. glaucus group,A. flavus. A, fumigatus, A. niger group,A. ochraceus group,A. ochraceus, Chœtomium sp.,Cladosporium cladosporioides. Cylindrocarpon sp.Doratomyces sp.,Fusarium sp.,F. moniliforme. F. oxysporum. F. solani, F. subglutinans. Marianaea elegans, Mortierella sp.,Mucor sp.,Myrothecium sp.,Penicillium sp.,Rhizopus sp.,Scopulariopsis brevicaulis. Sepedonium sp.,Stachybotrys chartarum. Stemphylium sp.,Torula humicola andTrichoderma viride were isolated.     

A study of fungal spore poupulations in the atmosphere of Kuwait

The fungal air-spora of Kuwait was investigated using the Petri-dish trapping technique. During the period from 1st April, 1974 to 30th June, 1975, a total of 3685 colonies were recorded from 2 % malt agar plates. Fifty-five genera and 116 species were identified. Alternaria occupied the first place in the order of percentage incidence, being represented by 18.3% of the entire catch, followed by Aspergillus (17.1%), Penicillium (14%), Cladosporium (13.6%), Drechslera (13.3%), and Ulocladium (7.1%). The widest spectrum of species was displayed by Aspergillus (possessed 19 species) followed by Penicillium (17 species), Alternaria (6 species), Drechslera (5 species), and Ulocladium (4 species). The monthly total number of fungi showed marked seasonal periodicity with the greatest number of colonies occurring in March–April and November. The lowest counts were recorded in mid-summer (July) and mid-winter (January). Only 220 colonies and 17 species of thermophilous fungi were reported from plates incubated at 45 °C. The most common species were Aspergillus fumigatus and A. terreus.     

Non-dermatophytic Molds as Agents of Onychomycosis in Izmir,Turkey – A Prospective Study

The purpose of this study was to determine the prevalence of causative non-dermatophytic filamentous fungi in onychomycosis. Totally 1,222 (1,222 × 3 = 3,666) samples of nail scrapings from 1,146 patients (from 76 patients two specimens: both from finger- and toe-nails) with prediagnosis of onychomycosis sent to the Mycology Laboratory from the Clinic of Dermatology, Ege University Hospital, Izmir, Turkey, July 2001–December 2003, were prospectively studied with conventional mycological procedures. The set criteria for the diagnosis of onychomycosis due to non-dermatophytic molds were: (1) Observation of fungal elements in 15% KOH-preparations made from nail scrapings, (2) growth of the same mold in all three consecutive cultures of the specimens taken three times from the same patient with one-week intervals, (3) no growth of a dermatophyte or yeast in three consecutive cultures. As agents of onychomycosis molds were detected in 33 (9%), dermatophytes in 175 (48%), yeasts in 150 (41%), and mixed (two different fungi) in 8 (2%) patients. In cases of mold onychomycosis, 11 (33%) had finger-nail and 22 (67%) toe-nail infection; 25 (76%) were female and 8 (24%) male; and 27 (82%) were above 40 years of age. The agents of mold onychomycosis, in order of frequency, were Aspergillus niger (7), Acremonium spp. (6), Fusarium spp. (6), Ulocladium spp. (4), sterile mycelia (2), Alternaria sp. (1), Aspergillus flavus (1), Aspergillus fumigatus (1), Aspergillus terreus (1), Cladosporium sp. (1), Paecilomyces spp. (1), Scopulariopsis sp. (1) and Trichoderma sp. (1). In conclusion, this study showed that non-dermatophytic molds were responsible for nearly 10% of onychomycoses cases attending the dermatology outpatient clinic of a university hospital in Izmir, Turkey. Since molds are common contaminants in the laboratory, cultures from consecutively taken nail scrapings should be made and carefully evaluated in order to diagnose a “mold onychomycosis”.     

Colonization capacity of the soil micro-organisms in relation to fungistasis

Summary Evans' tube and cellophane agar-disc techniques were used to evaluate the colonization capacity of the soil microbes and fungistasis, respectively. Spore germination ofAlternaria tenuis, Curvularia geniculata, Helminthosporium rostratum andPestalotia sp. was studied on a natural soil. Aspergillus fumigatus was the pioneer colonizer, followed in the order of decreasing colonizing capacity by,A. flavus, A. niger, A. terreus andPenicillium nigricans. Sensitivity of test spores to soil fungistasis declined in the order,C. geniculata → Pestalotia sp. →A. tenuis → H. rostratum. The fungistatic level of the soil samples decreased gradually from side arms 1 to 7 of the Evans tube, inoculated with unsterilized soil from one end. Studies conducted with an inoculum of four soil fungi (A. fumigatus, A. flavus, A. niger, andP. nigricans) as well as those involving fumigation of Evans tube by CS₂ also proved the greater colonizing capacity ofA. fumigatus. The level of fungistasis followed closely the colonization pattern of this fungus. When added individually to the soil, all fungi were able to inhibit spore germination of test fungi to some degree or other.     

Differences in the microflora of scarified and unscarified seeds ofKarwinskia humboldtiana (Rhamnaceae)

Seeds ofKarwinskia humboldtiana obtained from a 1997 collection in the locality of Villa de García Nuevo (León, Mexico) were contaminated with spores of filamentous fungi, bacteria and yeasts. The concentration of microorganisms in unscarified seeds ranged from 3.0×10³ to 7.5×10³ CFU/g. Predominant were bacterial isolates of the generaAeromonas sp.,Bacillus, andPseudomonas; from filamentous fungi were identifiedAlternaria, Aspergillus niger, Cladosporium sp.,Fusarium sp.,Mucor sp.,Penicillium commune, Trichothecium sp.; from yeastsRhodotorula sp. andSaccharomyces cerevisiae. Seed scarification significantly reduced the microbial contamination. Of the original fungal isolates, only two were identified on scarified seeds,viz. Cladosporium sp. andSaccharomyces cerevisiae; although a relatively high incidence of a unidentifiable ofPenicillium sp. was found, the bacterial spectrum was not altered. Treatment of scarified seeds with Vitavax 200 WP and Pomarsol Forte 80 WP (3 mg/g seeds) augmented germination by 10–19% compared to treated unscarified seeds, and by 16–31% compared to untreated unscarified seeds.     

Daily and seasonal variations ofAlternaria andCladosporium airborne spores in León (North-West, Spain)

The spores ofAlternaria andCladosporium are present throughout the year in the atmosphere of León (NW Spain), although they show an important seasonal variation. To understand the relationship between the number of spores and climatic factors,Alternaria andCladosporium spores counts for January 1994 to December 1995 were examined by means of correlation analyses. The results of weekly samples of both years showed that the spores concentration of two taxa are significantly and positively correlated with maximum and minimum temperature and sunshine hours and negatively with relative humidity. The statistical analysis of daily samples generally showed the same results. In the hourly distribution of spore concentrations we can see a similar behaviour ofAlternaria andCladosporium, with most spores collected in the 12–14 h period.     

Two new species ofPseudopetigurus (Trilobita) from the Arenig — Llanvirn of South China

Two new species ofPseudopetigurus are described from northern Hunan Province, China.P. deprati n. sp. is abundant in the upper Zitai Formation (upper Arenig) of Shuanghong, Baiyun, Shimen County, andP. expansus n. sp. occurs throughout the Jiuxi Formation (Arenig — lower Llanvirn) of Panjiazui, Taoyuan County. These formations represent outer shelf facies from the Jiangnan Transitional Belt of the South China Plate, close to the southeast margin of the Yangtze Platform. BothP. deprati andP. expansus show similarities to Upper Ordovician species ofPseudopetigurus recorded from South China. Other species ofPseudopetigurus are known almost entirely from cranidia, but pygidia and librigenae are also described for both new species, and the first known hypostome assigned to the genus is described forP. expansus. These two species are ecologically important members of offshore trilobite faunas during the Arenig — Llanvirn and define the outer shelfPseudopetigurus Association of South China. Despite being a deep-water genus,Pseudopetigurus is biogeographically restricted to the western margin of the Gondwanan craton and neighbouring regions of Baltica.      

Ulocladium systematics revisited: phylogeny and taxonomic status

The genus Ulocladium represents phaeodictyosporic Hyphomycetes that produce conidia that are essentially obovoid in shape. Previous molecular studies that included Ulocladium and related taxa in Alternaria, Embellisia, and Stemphylium revealed a conflict between morphology and phylogeny, and Ulocladium was supported as polyphyletic with a paraphyletic core group. Moreover, the genus consistently resolved within a larger Alternaria/Ulocladium clade, resulting in paraphyly of Alternaria and questions as to the taxonomic status of Ulocladium. In the present study, 13 Ulocladium species and three genetic loci were included for a more comprehensive systematic analysis of the genus than had previously been conducted. Total genomic DNA was extracted from representative taxa and sequences were determined for the nuclear internal transcribed spacer region, including the 5.8S rDNA gene, and the protein-coding genes glyceraldehyde-3-phosphate dehydrogenase and Alt a1. Subsequent phylogenetic analyses based on maximum parsimony and Bayesian methods included related Alternaria, Embellisia, and Stemphylium spp. Results supported previous findings of polyphyletic and paraphyletic relationships of Ulocladium among other taxa. Ten Ulocladium species clustered into a core Ulocladium clade and all taxa possessed the key diagnostic feature of Ulocladium, namely, conidia essentially obovoid in shape. However, A. cheiranthi and E. indefessa also clustered within this group with high bootstrap support but did not posses this diagnostic feature. This paraphyletic clade resolved basal to the core Alternaria clade with high bootstrap support, unlike previous studies in which its position was imbedded within the primary Alternaria clade. Thus, the status of the genus as an independent lineage and a unique taxon is strongly supported. As previously reported, U. alternariae and U. oudemansii, which posses the key conidium characteristics of Ulocladium, clustered as a separate clade sister to the core Ulocladium clade. Further studies are necessary to determine if these taxa represent an independent lineage or share a common ancestor with other Ulocladium species. Obovoid conidia were poorly represented in the isolate of U. lanuginosum that was included in these analyses (the only U. lanuginosum isolate currently available), and the isolate resolved as A. radicina based upon all three loci sequenced. Based upon these data and the origin of the isolate, which was originally deposited as A. malvae, a reassessment of its identity is supported.     

Lower Permian Bryozoa from Ellesmere Island (Canada)

A collection of fossils sampled during the 1898–1902 expedition of theFram to the Canadian Arctic Islands includes abundant bryozoans from the Lower Permian (Artinskian) Great Bear Cape Formation of Ellesmere Island. From this material a new genus with one new species —Nansenopora peculiaris n. gen., n. sp. — as well as three new species —Streblotrypella arctica n. sp.,Phragmophera patricki n. sp. andKallodictyon spinatum n. sp. — are described. Furthermore, the speciesUlrichotrypa ramulosa Bassler, 1929 is reported for the first time from the Lower Permian of the Arctic region.     

Studies on fungi associated with tomato fruits and effectiveness of some commercial fungicides against three pathogens

Among 39 fungal species and one variety belonging to 16 genera isolated from 116 healthy tomato fruits collected from markets in Assiut, Egypt, during 1994,Aspergillus niger was found to be the most prevalent, being isolated from 84.6% of the samples. Of 11 species recovered from 156 diseased tomatoes,Alternaria alternata, Rhizopus stolonifer andA. niger were the most common and isolated from 52.7%, 35.9% and 25.0% of the samples, respectively. Experiments for comparison of the effects of medium containing tomato juice with synthetic medium on the mycelial growth of nine fungal species indicated that, the tomato juice medium was more suitable for growth of all fungal species. The effect of five commercial fungicides and sodium hypochlorite when applied as post-harvest dips after inoculation was studied in laboratory againstA. alternata, A. niger andR. stolonifer. At 10 and 100 µg ml^–1, neither of the fungicide caused a noticeable inhibition ofAlternaria rot. At 1000 µg ml^–1 benlate, rovral and sumisclex completely preventedAlternaria andAspergillus rot, whereas cuprosan and ridomil were ineffective against rotting caused byA. niger. Rhizopus rot development was inhibited by over 50% with one treatment only (rovral at 1000 µg ml^–1). Sodium hypochlorite has good curative properties against fruit rots especially those caused byA. alternata andA. niger.     

Fungicide inhibition of aflatoxins,diacetoxyscirpenol and zearalenone production

The effect of fungicides on the production of aflatoxin byAspergillus flavus IMI 89717, diacetoxyscirpenol and zearalenone byFusarium graminearum was studied. In a yeast extract - sucrose medium, dicloran, iprodione and vinclozolin fungicides significantly inhibited mycelial growth ofA. flavus at 250 ppm and significantly decreased aflatoxin production at 100, 250 and 500 ppm, respectively. In potato — dextrose broth, these fungicides diminished the mycelial growth ofF. graminearum and production of diacetoxyscirpenol and zearalenone at 100 ppm. Sensitivity of toxigenic mycelia to fungicides increased approximately five-fold in a yeast extract-starch medium with an appreciable reduction in sugar uptake andα-amylase activity.     

The role of meteorological factors in determining the annual variation ofAlternaria andCladosporium spores in the atmosphere of Palencia, 1990–1992

A study was made of the daily content ofAlternaria andCladosporium spores in the atmosphere in Palencia city (Spain) for three consecutive years 1990–1992.Alternaria andCladosporium represented 55% of the total identified spores, presenting an annual distributional pattern of which the maximum values were reached in summer. Multiple regression analyses showed a positive correlation between minimum temperature andCladosporium spore concentrations, while forAlternaria there was a positive correlation with maximum temperature and a negative correlation with precipitation. Duncan's multiple range test among means ofAlternaria data indicated that winds coming from a northeasterly direction were associated with significantly higher concentrations the spores.     

Intraspecific invariability of the internal transcribed spacer region of rDNA of the truffleTerfezia terfezioides in Europe

ITS regions (internal transcribed spacers—ITS1 andITS2—with the 5.8S gene of the nuclear rDNA) of 25 fruit body samples ofTerfezia terfezioides, originating from Hungary and Italy, were compared. The amplification and sequencing of the ITS region was successful with both theITS1-ITS4 andITSIF-ITS4 primer pairs. No differences of the restriction fragment length polymorphism profiles were detected among 19 samples collected in one place at the same time. The sequences of the ITS region of 9 samples collected in different localities were highly invariable, differing in only two bases. Thus the intraspecific homogeneity of the ITS region seems to be an important species-specific characteristic ofT. terfezioides in contrast to otherTerfezia species. As the samples of the species were collected from different and distant localities of Europe, the ITS sequence ofT. terfezioides can be considered a very conservative, reliable molecular marker of the fungus. *** DIRECT SUPPORT *** A00EN076 00008     

Mapping of theHox-3.1 andMyc-1.2 genes on chromosome 15 of the mouse by restriction fragment length variations

Restriction endonuclease fragment length variations (RFLV) were detected by use of the cDNA probeHox-3.1 for the homeo box-3.1 gene and also thec-myc oncogene probe for exon 2. RFLV ofHox-3.1 were found inHindIII restriction patterns, and RFLV of theMyc-1.2 gene inEcoRV patterns. From the RFLV, theHox-3.1 andMyc-1.2 genes were mapped on chromosome 15. Three-point cross test data showed that the frequency of recombination is 26.4% betweenMyc-1.2 andGpt-1, 30.2% betweenGpt-1 andGdc-1, and 9.4% betweenGdc-1 andHox-3.1. The following order of these genes is proposed,Myc-1.2—Gpt-1—Gdc-1—Hox-3.1. All laboratory strains carry theHox-3.1 ^a andMyc-1.2 ^a alleles. Among strains of wild origin,domesticus strains carry only theHox-3.1 ^a andMyc-1.2 ^a alleles, as do the laboratory strains. One strain ofbrevirostris carries theHox-3.1 ^a andMyc-1.2 ^b alleles. Other wild subspecies from Europe and Asia,M. m. musculus, M. m. castaneus, M. m. molossinus, Chinese mice of wild origin, andM. m. yamashinai carry theHox-3.1 ^b andMyc-1.2 ^b alleles.     

In vitro screening of seaweed extract on the proliferation of mouse spleen and thymus cell

A total number of 31 types of seaweed were assessed with regard to their effects on the proliferation of mouse spleen and thymus cells in a culture, using an MTT reduction assay. Acetone:dichloromethane (1∶1) extracts of three seaweed plants:Derbesia marina, Sargassum sp., andHisikia fuziformis, exhibited significantly positive effects on the survival of mouse spleen and thymus cellsin vitro. The acetone: dichloromethane (1∶1) extracts ofSargassum sp., in particular, much more potent effects on thymus cell activation than did any of the other types of seaweed. However, the methanol extracts ofSargassum ringgoldianium andChondrus crispus exerted a stimulatory influence only on the proliferation of mouse spleen cells, whereas the methanol extracts ofGrateloupia lanceolata exhibited significant cell proliferation properties in both spleen and thymus cells.     

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.     

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.     

Incidence, Aggressiveness and In Planta Interactions of Botrytis cinerea and other Filamentous Fungi Quiescent in Grape Berries and Dormant Buds in Central Washington State

Recovery of quiescent filamentous fungi from non‐symptomatic grape berries and dormant buds demonstrated dominance of Alternaria, Aureobasidium, Cladosporium, Ulocladium and other dematiaceous hyphomycetes. Up to 78% of berries contained fungi prior to harvest. Botrytis cinerea was recovered from 0.2 to 0.5% of surface‐disinfested berries just subsequent to fruit set, and 1.6–4.8% of surface‐disinfested, over‐wintered dormant buds. In laboratory inoculations of mature grape berries with strains of Alternaria, Aureobasidium, Cladosporium, Ulocladium and Botrytis, only the latter was aggressive in rotting berry fruits. Inoculations with B. cinerea alone and in combination with strains of Alternaria, Aureobasidium, Cladosporium and Ulocladium recovered from grape demonstrated that prior occupation of wound sites by the latter fungi resulted in reduced lesion size compared to inoculation with B. cinerea alone.     

Airborne fungal spores colonising marbles exposed in the terrace of Messina Museum, Sicily

Studies were carried out on the air and on Carraramarble blocks located in the terrace of MessinaMuseum, in order to know the likelihood of airbornefungal spores coming into contact with and colonisingtridimensional objects. Our results showed there were not significantdifferences between airborne fungi circulating inspring and in autumn; Aspergillus, Penicillium,Fusarium, Alternaria, Cladosporium,Ulocladium, Aureobasidium, Phoma were themost common isolates. However, only few species wereable to settle on the marble surfaces as demonstratedby their isolation after 2 and 6 years of exposition.     

Enhancement of tessaric acid production in Tessaria absinthioides cell suspension cultures

The total production of the sesquiterpene tessaric acid (TA) by cell cultures of Tessaria absinthioides at day 25 of the culture period reached 0.086 mg g⁻¹ DW, with intracellular accumulation accounting for 0.059 mg g⁻¹ DW. Dimethylsulfoxide-induced permeabilization of the cells effected both total production and extracellular accumulation of the sesquiterpene to reach levels of 148% and 271%, respectively. Cultures treated with elicitor preparations of Verticillum sp., Monodyctis cataneae, Acremonium sp., and Aspergillus niger produced TA at levels of 281%, 197%, 149%, and 139%, respectively. Treatment of cell suspension cultures with cis-(-)-jasmonic acid (5 μM) increased production to 267%, whereas jasmonic acid pretreatment and subsequent elicitation raised external tessaric acid to 702%. Received: 16 December 1998 / Revision received: 02 November 1999 / Accepted: 19 November 1999