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Phylogenetic relationships in Termitomyces (Family Agaricaceae) based on the nucleotide sequence of ITS: a first approach to elucidate the evolutionary history of the symbiosis between fungus-growing termites and their fungi.

Termitomyces constitutes a very poorly known genus of fungi whose essential characteristic is that all representatives of the genus are cultivated by termites (Macrotermitinae) in their nest and that all the fungi cultivated by termites belong to this genus. For the first time, the phylogenetic relationships of several African Termitomyces species was studied by the sequencing of their internal transcriber spacer region (ITS1--5.8S--ITS2). It appeared that this group is clearly monophyletic and belongs to the Tricholomataceae family. The total homology of the ITS zone of several Termitomyces symbionts of different termite genera indicated that the specific diversity of this group is in fact less important than previously supposed. Finally, the comparison between the Termitomyces phylogenetic tree and the taxonomic tree of Macrotermitinae showed that if for certain genera the hypothesis of termite/fungus coevolution is acceptable, it should not be applied for all symbiosis.

Agaricales↗

Degradation of xenobiotics by white rot fungi.

White rot fungi such as P. chrysosporium degrade the nonrepeating, nonstereoselective, insoluble polymer lignin under conditions of nutrient limitation. The attack on lignin principally involves extracellular peroxidases (ligninases) and hydrogen peroxide. Hydroxyl radicals may also make a significant contribution. The ligninolytic system lends itself to the degradation of xenobiotics, since these often have limited solubility in water and are not readily available in soil to intracellular metabolism. A nonspecific attack should proceed at a rate independent of the target's concentration and the fungal system would be expected to remediate soil contaminated with a mixture of compounds. This contrasts with the need for induction and problems with simultaneous metabolism encountered with bacterial inoculation. The P. chrysosporium system has been found active against such diverse substrates as DDT, lindane, PCBs, TNT and crystal violet, with substantial mineralization in many cases. Some like biphenyl and triphenylmethane dyes are structurally related to lignin substructures while others bear groups such as nitro (TNT) or halogen (PCP) that are absent from the natural polymer. The fate of transformed targets varies: pentachlorophenol is incorporated into soil organic matter as a result of fungal ligninase action, whereas highly lipophilic Aroclor PCBs are converted to water-soluble metabolites. Normally less toxic intermediates are generated: for example, with benzo[a]pyrene, mutagenic arene oxides do not appear in the white rot fungal system. In certain cases, purified ligninases were also active in degrading pollutants such as PCP, benzo[a]pyrene or triphenylmethane dyes. Methods of optimizing ligninase activity in fungal reactors have been described, such as the addition of surfactants and veratryl alcohol to the medium. It remains to be seen how molecular biology can provide further advances in maximizing the bioremediating activity of white rot fungi applied to contaminated soil.

Basidiomycota↗

Phylogeny of Alternaria fungi known to produce host-specific toxins on the basis of variation in internal transcribed spacers of ribosomal DNA.

The internal transcribed spacer regions (ITS1 and ITS2) of ribosomal DNA from Alternaria species, including seven fungi known to produce host-specific toxins, were analyzed by polymerase chain reaction-amplification and direct sequencing. Phylogenetic analysis of the sequence data by the Neighbor-joining method showed that the seven toxin-producing fungi belong to a monophyletic group together with A. alternata. In contract, A. dianthi, A. panax, A. dauci, A. bataticola, A. porri, A. sesami and A. solani, species that can be morphologically distinguished from A. alternata, could be clearly separated from A. alternata by phylogenetic of the ITS variation. These results suggest that Alternaria pathogens which produce host-specific toxins are pathogenic variants within a single variable species, A. alternata.

Alternaria↗

Transformation of seven species of filamentous fungi using the nitrate reductase gene of Aspergillus nidulans.

A gene transfer system originally developed for Fusarium oxysporum has been applied to seven species of filamentous fungi of agricultural and industrial importance. This transformation system relies on the selection of mutants deficient in nitrate reductase by positive screening. Such mutants were recovered easily in all the fungi tested--without mutagenic treatments--through their resistance to chlorate. They were transformed by a plasmid vector (pAN301) carrying the Aspergillus nidulans wild-type gene (niaD). Transformation frequencies ranged from one to ten transformants/micrograms plasmid DNA. The general properties of the transformants were analyzed. Most of them are mitotically stable, and the integration of the vector into the host genome frequently occurred in a tandem fashion.

Aspergillus nidulans↗

Isolation of keratinophilic fungi from soil in Malaysia.

Two hundred and thirty soil samples from different localities were examined for the presence of geophilic keratinophilic fungi. Six species namely Microsporum gypseum--34 isolates, Chrysosporium keratinophilum--29, C. tropicum--20, Keratinophyton terreum--4, Trichophyton terrestre--8 and Chrysosporium species--3--were isolated. Most of these fungi were recovered from garden, field and river bank soil. The importance of these findings is briefly discussed.

Ascomycota↗

Keratinophilic fungi on the hair of cows, donkeys, rabbits, cats, and dogs from the West Bank of Jordan.

The mycoflora of hair of 111 cows, donkeys, rabbits, cats, and dogs were analysed and the frequency of occurrence and the relative importance value of the different fungal species isolated were calculated. Total numbers of species 56, 45, 48, 23, and 11 were recovered from cows, donkeys, rabbits, cats, and dogs respectively. The majority of the keratinophilic fungi isolated were either well-known mycotic agents or have been recovered from various animal and human lesions. These comprised 87.8%, 66.7%, 61.4%, 59.3%, and 55.2% of the hair mycoflora in cats, dogs, rabbits, cows, and donkeys respectively. Seven species of dermatophytes were isolated: Trichophyton mentagrophytes, T. verrucosum, Microsporum gypseum, M. nanum, T. ajelloi, and M. canis. Cats harboured the largest number of dermatophytes (5 species), followed by cows (4 spp), rabbits (3 spp), and donkeys and dogs (1 sp). The role of these animals in the persistence and transmission of pathogenic fungi is discussed.

Animals↗

Osmophilism in foodgrain storage fungi.

A comparative study of different species of storage fungi isolated from rice showed them to be truly osmophilic rather than osmotolerant. Their osmophilism was of a high order, their optimum sucrose requirement being in the range of 50% to 75% w/v in the medium. However, only Aspergillus glaucus was obligately osmophilic. A sucrose concentration of 50% w/v in Czapek-Dox agar was concluded to be the most suitable for the isolation of these different fungi from foodgrains.

Aspergillus↗

The application of immunoperoxidase staining for the detection of causative fungi in tissue specimens of mycosis I.

This study was performed in order to identify the fungi of four species (Aspergillus fumigatus, Fusarium anthophilum, Candida albicans, Cryptococcus neoformans) in formalin-fixed, paraffin-embedded tissue sections by the indirect method of immunoperoxidase staining. Mature albino rabbits were immunized by formalin-killed organisms. The antibodies were prepared by precipitation at a 50% saturation of ammonium sulfate and were checked for cross-reactivities by Ouchterlony's double immunodiffusion and precipitin test. The immunoperoxidase staining was applied to the paraffin-embedded tissue sections of infected mice, human autopsy and biopsy specimens. Although each fungus was stained clearly the cell wall, cross-reactivities appeared among them, however it was possible to identify four fungi by absorption and dilution of the antisera.

Animals↗

Survey of aflatoxin-producing fungi in certain fermented foods and beverages in Thailand.

Aflatoxin-producing fungi were found in fermented foods and beverages: fermented rice (kaomak), soybean sauce (taotjo), peanut butter, soy sauce (shoyu), Thai red and white wine, and rice sugar wine. These foods were extracted directly and tested for aflatoxins by thin-layer chromatography (TLC) and high pressure liquid chromatography (HPLC). Four strains of aflatoxin-producing fungi were isolated from peanut butter, taotjo, and shoyu. Direct extracts of 10% of the peanut butters tested and 5% of the kaomak tested contained large amounts of aflatoxins. The HPLC procedures used in this experiment utilized chloroform-ethyl acetate (3:1).

Aflatoxins↗

Dermatophytes and other associated fungi isolated from ringworm lesions of camels.

Among 75 camels showing skin lesions, 48% were positive for fungal infection. The younger individuals were more susceptible to this infection. Sixteen species belonging to nine genera of keratinophilic and cycloheximide-resistant fungi were recovered from diseased camels. Trichophyton, Microsporum and Chrysosporium were the most common genera. T. verrucosum appeared to be the main cause of ringworm in small camels while T. mentagrophytes infected older ones. Camel skin presents a suitable habitat for the growth of some dermatophytes and other potentially pathogenic fungi.

Age Factors↗

Production of lytic enzymes by Trichoderma spp. and their effect on the growth of phytopathogenic fungi.

The production of beta-1,3-glucanases and chitinases by three strains of Trichoderma in submerged cultures was determined. The synthesis of enzymes was induced by cell wall biopolymers of phytopathogenic fungi (Botrytis cinerea, Fusarium culmorum and F. oxysporum). T. hamatum produced the highest beta-1,3-glucanase activity; the most effective inducer of enzyme synthesis was the biomass of F. oxysporum. All examined strains of Trichoderma inhibited phytopathogen growth in biotic tests. The diffusion tests showed that the lytic enzymes take part in growth inhibition of phytopathogenic fungi.

Botrytis↗

Free and bound magnesium in fungi and yeasts.

The content of total, bound and osmotically free magnesium was estimated in various fungi and in the yeast Saccharomyces cerevisiae. Total magnesium increases at lower growth rates of Endomyces magnusii and Penicillium chrysogenum 140A as well as during the logarithmic stage of growth of Penicillium chrysogenum Q-176. The binding of magnesium requires orthophosphate, decreasing during lack of external phosphate when the intracellular concentration of free magnesium rises. The fungi were found to contain a novel form of bound magnesium, a polymeric magnesium orthophosphate (PO Mg), which appears to take part in the control of free magnesium level in Penicillium chrysogenum Q-176. The level of free magnesium is proportional to the growth rate of Endomyces magnusii and Penicillium chrysogenum Q-176 and 140A. Total, as well as free, magnesium changes less than three-fold as external Mg concentration is changed 13,000-fold. The magnesium is taken up against concentration gradients of 1 : 25 to 1 : 1300, the metal being distributed non-uniformly in the cells of Saccharomyces cerevisiae.

Ascomycota↗

Image processing by microcomputer on ultrastructure of griseofulvin-resistant fungi in favus.

The ultrastructure of 5 griseofulvin-resistant fungi of favus was studied by image processing with microcomputer. It was found that the cell walls of the fungi consisted of 8 layers, and the inner layer containing cytoplasm was loose. It was also found that all structures within the cytoplasm possessed a 1-3 layers integral envelope with chromatins in the nucleus. These might be contributing factors in the development of resistance to griseofulvin. This multiple-layered, thick cell wall might act as a barrier responsible for the impermeability of the cell to griseofulvin.

Drug Resistance, Microbial↗

Incorporation of cadmium into proteins in a cadmium tolerant fungi.

Aspergillus carbonarius and a strain of Penicillium, a cadmium tolerant fungi, are able to metabolize cadmium chloride up to 2% (w/v). Their amino acids analysis on cadmium free and cadmium chloride containing media indicated certain disorders in their metabolic activities. Cystathionine was only detected in both fungi in the presence of cadmium chloride. However, cadmium was incorporated into several types of low and high molecular weight proteins. The amino acids hydrolyzates of a cadmium containing protein are characterized by the presence of high levels of sulfur amino acids; cysteine and methionine. Ethylasparagine was detected in the hydrolyzate of that cadmium containing protein as well.

Amino Acids↗

Differentiation of anaerobic polycentric fungi by rDNA PCR-RFLP.

The suitability of restriction fragment length polymorphism (RFLP) analysis of the ribosomal DNA cluster for discriminating two genera of anaerobic polycentric fungi, Orpinomyces and Anaeromyces, was determined. Three PCR-amplified DNA fragments--nuclear small subunit (SSU; 18S rDNA), the nuclear large subunit (LSU; 28S rDNA) and internal transcribed spacer (ITS)--were restricted with endonucleases AluI, DraI, HinfI and MboI. Although the SSU DNA fragment could be restricted successfully by all four enzymes, no differences were observed between restriction patterns of Orpinomyces and Anaeromyces. The most polymorphic restriction pattern between Orpinomyces and Anaeromyces resulted from cleavage of LSU rDNA fragments cut by AluI and HinfI and ITS fragment cut by DraI and HinfI. Genus-specific RFLP patterns were determined for Orpinomyces and Anaeromyces genera; the results showed that the PCR-RFLP analysis of rDNA offers an easy and rapid tool for differentiation of two polycentric genera of anaerobic fungi, which could be hardly separated on the basis of morphology.

Classification↗

Fungicidal effect of resveratrol on human infectious fungi.

Resveratrol, a phenolic antioxidant found in grapes, has been known to mediate various biological activities on the human body. In the present study, we tested the antifungal activity of resveratrol against human pathogenic fungi before carrying out further studies to elucidate the antifungal mechanism(s) of resveratrol. Resveratrol displayed potent antifungal activity against human pathogenic fungi at concentration levels of 10-20 microg/mL. Furthermore, time-kill curve exhibited fungicidal effect of resveratrol on C. albicans, but the compound had no hemolytic activity against human erythrocytes. The destruction of C. albicans cells by resveratrol was confirmed by scanning electron microscopy. These results suggest that resveratrol could be employed as a therapeutic agent to treat fungal infections of humans.

Antifungal Agents↗

Reducing dust, lead, dust mites, bacteria, and fungi in carpets by vacuuming

Old carpets may be reservoirs of dust, lead (Pb), and dust mite allergen. The purpose of this study was to determine if the dust, Pb, dust mite allergen, bacteria, and fungi on the surface of carpets could be reduced by 90% in 1 week with the use of a Hoover Self Propelled Vacuum with Embedded Dirt Finder (HSPF). A high-volume surface sampler (HVS3) was used to measure surface dust and pollutants before and after the use of the HSPF to remove deep dust in carpets 12 to 20 years old in nine middle-class homes and two small offices. The minimum, median, and maximum surface loading for the first and final samples are as follows: first fine dust loading: min 0.32 g/m2, max 14.4 g/m2, median 1.30 g/m2; final fine dust loading: min 0.019 g/m2, max 0.289 g/m2, median 0.102 g/m2; first Pb loading: min 38 &mgr;g/m2, max 3,871 &mgr;g/m2, median 471 &mgr;g/m2; final Pb loading: min 13 &mgr;g/m2, max 2,023 &mgr;g/m2, median 86 &mgr;g/m2; first mite loading: min 0.11 &mgr;g/m2, max 12. 88 &mgr;g/m2, median 2.82 &mgr;g/m2; final mite loading: min 0.06 &mgr;g/m2, max 3.67 &mgr;g/m2, median 0.28 &mgr;g/m2. Data were insufficient to determine if the loadings of bacteria and fungi were reduced a similar amount. Six to 45 min/m2 of vacuuming with the HSPF removed deep dust from these carpets. The median surface loadings of fine dust, Pb, and dust mite allergen in these 11 carpets were reduced by 91, 82, and 94%, respectively, in 1 to 15 h of vacuuming. The loading of the deep dust collected with the HSPF ranged from 8 to 171 g/m2 with a median of 66 g/m2.

Journal Article↗