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PubMed2004

Volumetric assessment of airborne fungi in two sections of a rural indoor dairy cattle shed.

There is increasing concern about the exposure to fungal aerosols in occupational environments and associated respiratory allergic diseases and asthma. A large number of people work in cattle sheds around the world, pulmonary function impairments and higher frequency of respiratory symptoms have been reported in dairy farmers; however, it appears that adequate information on the fungal aerosols from the cattle sheds are largely lacking. Volumetric assessment of airborne culturable and nonculturable fungal spores was performed in two sections of a large rural indoor cattle shed of West Bengal, India for 2 consecutive years. An Andersen Two Stage Viable Sampler was used for sampling culturable fungi and a Burkard Personal Slide Sampler was used to collect the total airborne fungal spores including both the culturable and nonculturable types. A total of 31 spore types and 35 types of viable colony-forming units were recorded. Average concentration range of total fungal spores was 233-2985/m(3) and concentration of viable colony-forming units ranged between 165 and 2225 CFU/m(3). Burkard Sampler showed higher frequencies of Aspergilli/Penicilli, Cladosporium, Alternaria, and smut spores. Andersen Sampler showed the prevalence of Aspergillus niger, Aspergillus flavus and Cladosporium cladosporioides colonies. Some recorded fungi were earlier reported as allergenic, toxic, and pathogenic for occupational workers as well as cattle population. Higher concentration levels of airborne total and culturable fungal spores were recorded during the months of November through February (winter) and June through September (late summer and rainy season).

Air Pollution, Indoor↗

PubMed1999

A rapid and efficient method for growth measurement of filamentous fungi.

Growth of filamentous fungi may be measured very efficiently using 96 well microtiter plates and a microplate reader. The relationship between Absorbance reading at 630 nm and dry weight is linear. This relationship seems to be similar for different fungi with a slope of about 4.2 mg/ml dry weight per Absorbance unit. The 8 wells in each column were used as parallels.

Colony Count, Microbial↗

PubMed2002

Filamentous fungi as cell factories for heterologous protein production.

Filamentous fungi have been used as sources of metabolites and enzymes for centuries. For about two decades, molecular genetic tools have enabled us to use these organisms to express extra copies of both endogenous and exogenous genes. This review of current practice reveals that molecular tools have enabled several new developments. But it has been process development that has driven the final breakthrough to achieving commercially relevant quantities of protein. Recent research into gene expression in filamentous fungi has explored their wealth of genetic diversity with a view to exploiting them as expression hosts and as a source of new genes. Inevitably, the progress in the 'genomics' technology will further develop high-throughput technologies for these organisms.

Fermentation↗

PubMed2003

Using fungi and yeasts to manage vegetable crop diseases.

Vegetable crops are grown worldwide as a source of nutrients and fiber in the human diet. Fungal plant pathogens can cause devastation in these crops under appropriate environmental conditions. Vegetable producers confronted with the challenges of managing fungal pathogens have the opportunity to use fungi and yeasts as biological control agents. Several commercially available products have shown significant disease reduction through various mechanisms to reduce pathogen development and disease. Production of hydrolytic enzymes and antibiotics, competition for plant nutrients and niche colonization, induction of plant host defense mechanisms, and interference with pathogenicity factors in the pathogen are the most important mechanisms. Biotechnological techniques are becoming increasingly valuable to elucidate the mechanisms of action of fungi and yeasts and provide genetic characterization and molecular markers to monitor the spread of these agents.

Anti-Bacterial Agents↗

PubMed2003

Influence of temperature, water activity and pH on growth of some xerophilic fungi.

The combined effects of water activity (aw), pH and temperature on the germination and growth of seven xerophilic fungi important in the spoilage of baked goods and confectionery were examined. Eurotium rubrum, E. repens, Wallemia sebi, Aspergillus penicillioides, Penicillium roqueforti, Chrysosporium xerophilum and Xeromyces bisporus were grown at 25, 30 and 37 degrees C on media with pH values of 4.5, 5.5, 6.5 and 7.5 and a range of water activities (aw) from 0.92 to 0.70. The aw of the media was controlled with a mixture of equal parts of glucose and fructose. Temperature affected the minimum aw for germination for most species. For example, P. roqueforti germinated at 0.82 aw at 25 degrees C, 0.86 aw at 30 degrees C and was unable to germinate at 37 degrees C. E. repens germinated at 0.70 aw at 30 degrees C, but at 25 and 37 degrees C, its minimum aw for germination was 0.74. C. xerophilum and X. bisporus germinated at 0.70 aw at all three temperatures. The optimum growth occurred at 25 degrees C for P. roqueforti and W. sebi, at 30 degrees C for Eurotium species, A. penicillioides and X. bisporus and at 37 degrees C for C. xerophilum. These fungi all grew faster under acidic than neutral pH conditions. The data presented here provide a matrix that will be used in the development of a mathematical model for the prediction of the shelf life of baked goods and confectionery.

Culture Media↗

PubMed2001

Antifungal activity of Artemisia annua endophyte cultures against phytopathogenic fungi.

Artemisia annua, well recognized for its production of antimalarial drug artemisinin, is seldom attacked by any of phytopathogenic fungi, which could be partially associated with the presence of endophytes. Present investigation is aiming at disclosing whether the endophytes inside A. annua produce antifungal substances. A total of 39 endophytes were isolated and fermented, and the ferment broth was evaluated in vitro for the antifungal activity against crop-threatening fungi Gaeumannomyces graminis var. tritici, Rhizoctonia cerealis, Helminthosporium sativum, Fusarium graminearum, Gerlachia nivalis and Phytophthora capsici. These plant pathogens are still causing wheat take-all, sharp eyespot, common rot, scab, snow mould, and pepper phytophthora blight, respectively. Out of 39 endophytes investigated, 21 can produce in vitro substances that are inhibitory to all or a few of the tested phytopathogens whereas the rest yielded nothing active. Moreover, the most active broth of endophyte IV403 was extracted with EtOAc and n-butanol, and comparisons of the antifungal activity of the extracts indicated that the major active metabolites were EtOAc-extractable.

Actinomycetales↗

PubMed2002

The role of fungi in the transfer and cycling of radionuclides in forest ecosystems.

Fungi are one of the most important components of forest ecosystems, since they determine to a large extent the fate and transport processes of radionuclides in forests. They play a key role in the mobilization, uptake and translocation of nutrients and are likely to contribute substantially to the long-term retention of radiocesium in organic horizons of forest soil. This paper gives an overview of the role of fungi regarding the transfer and cycling of nutrients and radionuclides, with special emphasis on mycorrhizal symbiosis. Common definitions of transfer factors, soil-fungus and soil-green plant, including their advantages and limitations. are reviewed. Experimental approaches to quantify the bioavailability of radionuclides in soil and potential long-term change are discussed.

Biological Availability↗

PubMed2002

Influence of nematophagous fungi, earthworms and dung burial on development of the free-living stages of Ostertagia (Teladorsagia) circumcincta in New Zealand.

Biological options for nematode parasite control are being sought, as the long-term efficacy of conventional anthelmintics comes increasingly under threat from drug-resistant parasites. Three biological methods with the potential to reduce pasture contamination by parasitic nematode larvae were examined: (a) killing of larvae developing in dung by nematophagous fungi; (b) removal of dung through earthworm ingestion; (c) burial of dung in soil as might occur through the action of dung beetles. Field trials with the test bio-control agents were carried out in autumn and spring by adding dung from sheep infected with Ostertagia (Teladorsagia) circumcincta to pots of ryegrass/white clover. The factorial treatment structure included five fungal treatments (individual applications of Duddingtonia flagrans, Monacrosporium gephyropagum and Harposporium helicoides, a combination of all the three fungi together and an untreated control), two dung burial treatments (dung buried or deposited on the soil surface) and two earthworm treatments (earthworms present or absent). D. flagrans and H. helicoides, individually or in combination, reduced recovery of infective stage larvae in experiment 1, while only H. helicoides reduced recovery in experiment 2. In both the experiments, dung burial increased the total number of larvae recovered, while the number of infective larvae were reduced by the action of earthworms. Increased recovery following burial, along with the fact that larvae moved rapidly from soil onto herbage, suggests that soil may provide a protective reservoir for infective larvae infesting herbage.

Animals↗

PubMed2000

Regulation of plasma membrane H(+)-ATPase in fungi and plants.

The plasma membrane H+-ATPase from fungi and plants is a proton pump which plays a key role in the physiology of these organisms controlling essential functions such as nutrient uptake and intracellular pH regulation. In fungal and plant cells the activity of the proton pump is regulated by a large number of environmental factors at both transcriptional and post-translational levels. During the last years the powerful tools of molecular biology have been successfully used in fungi and plants allowing the cloning of a wide diversity of H+-ATPase genes and rapid progress on the molecular basis of reaction mechanism and regulation of the proton pump. This review focuses on recent results on regulation of plasma membrane H+-ATPase obtained by molecular approaches.

Amino Acid Sequence↗

PubMed1978

Production of protein by fungi from agricultural wastes. I. Standardization of certain factors for maximum protein production.

Several fungi were screened for protein production in a simple medium using either sugarcane bagasse, wheat straw or cowdung as substrate. Out of them Rhizoctonia melongina, Pleurotus ostreatus, and Coprinus aratus proved promising and were taken up for further studies. Optimum conditions of temperature, incubation, and pH for maximum growth and protein production by the three fungi were then ascertained and standardized.

Agaricales↗

PubMed1978

Cellulose-decomposing fungi.

The present article gives a survey of the cellulose-decomposing fungi. It is concerned with the micro-organisms having the capability of degradating cellulose sources. It includes the factors influencing cellulose-decomposing fungi, cellulose-decomposing enzymes, mechanisms of degradation, and factors influencing the cellulolytic enzymes (cellulases).

Biodegradation, Environmental↗

PubMed2003

Expression of the red fluorescent protein DsRed-Express in filamentous ascomycete fungi.

The recently reported red fluorescent protein DsRed from the reef coral Discosoma sp. represents a new marker that has been codon-optimized for high expression in mammalian cells. To facilitate expression of DsRed in ascomycete fungi, we used the clone pDsRed-Express (Clontech) for constructing a plasmid vector, pPgpd-DsRed, containing the constitutive Aspergillus nidulans glyceraldehyde 3-phosphate (gpd) promoter. This vector was used for co-transformation of Penicillium paxilli, Trichoderma harzianum and Trichoderma virens (syn. Gliocladium virens) together with either pAN7-1 or gGFP, both containing a gene for hygromycin resistance for transformant selection. In addition, gGFP contains a green fluorescent protein (GFP) gene for expression in Ascomycetes. Expression of DsRed-Express was obtained in all three fungi, indicating that DsRed can be used as a highly effective vital marker in Ascomycetes. Dual marked transformants expressed both DsRed-Express and GFP in the same mycelium and were used for non-quantitative comparison of the intensity of the fluorescence using confocal laser scanning microscopy.

Gene Expression Regulation, Fungal↗

PubMed2003

Phylogeny of the Glomerales and Diversisporales (fungi: Glomeromycota) from actin and elongation factor 1-alpha sequences.

The arbuscular mycorrhizal (AM) fungi have been elevated to the phylum Glomeromycota based on a ribosomal gene phylogeny. In order to test this phylogeny, we amplified and sequenced small subunit ribosomal RNA (SSUrRNA), actin and elongation factor 1 (EF1)-alpha gene fragments from single spores of Acaulospora laevis, Glomus caledonium, Gigaspora margarita, and Scutellospora dipurpurescens. Sequence variation within and among spores of an isolate was low except for SSUrRNA in S. dipurpurescens, and the actin amino acid sequence was more conserved than that of EF1-alpha. The AM fungal sequences were more similar to one another than to any other fungal group. Joint phylogenetic analysis of the actin and EF1-alpha sequences suggested that the sister group to the AM fungi was a Zygomycete order, the Mortierellales.

Actins↗

PubMed1997

Degradation of the polyamine alkaloid aphelandrine by endophytic fungi isolated from Aphelandra tetragona.

Members of the genus Aphelandra (Acanthaceae) produce rare macrocyclic polyamine alkaloids which consist of spermine acylated with two units of 3-(4-hydroxyphenyl)prop-2-enoic acid. Endophytic fungi were isolated from roots and shoots of Aphelandra tetragona and tested for their ability to metabolize the main alkaloid aphelandrine, which accumulates exclusively in the roots of the plants. Several endophytes were able to metabolize aphelandrine but only root endophytes belonging to the Nectriaceae were good metabolizers. In addition, the endophytes were grown on an agar medium containing putrescine, spermidine, or spermine as the sole nitrogen source. All fungi were able to grow on putrescine, but only the good aphelandrine metabolizers grew well on spermidine or spermine. Acremonium sp. 15, one of the most active metabolizers, grew also on a medium containing aphelandrine as sole nitrogen source. A number of strains thought to be conspecific with Acremonium sp. 15 were also tested for their ability to metabolize aphelandrine. The ability of the endophytes to metabolize aphelandrine suggests an ecological adaptation of the symbionts to their host. The possibility of using the aphelandrine metabolism as a taxonomic character is briefly discussed.

Alkaloids↗

PubMed2003

In vitro susceptibility testing of filamentous fungi: comparison of Etest and reference M38-A microdilution methods for determining posaconazole MICs.

The performance of the Etest for posaconazole susceptibility testing of 72 isolates of filamentous fungi was assessed in comparison with the National Committee for Clinical Laboratory Standards (NCCLS) approved standard broth microdilution method (M38-A). The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48h at 35 degrees C. Etest MICs were determined with RPMI agar containing 2% glucose and were read after incubation for 48h at 35 degrees C. The isolates included Aspergillus fumigatus, A. flavus, A. nidulans, A. niger, A. versicolor, A. oryzae, A. terreus, Cladosporium spp., Curvularia sp., Exophiala sp., Fusarium spp., Paecilomyces spp., Pithomyces sp., Penicillium spp. and Scedosporium apiospermum. Overall agreement between Etest and microdilution MICs was 84% for Aspergillus spp. and 100% for the less common opportunistic molds, with the exception of Penicillium spp. (67%). Where a discrepancy was observed between Etest and the reference method, the Etest tended to give lower values. The Etest method using RPMI agar appears to be a useful method for determining posaconazole susceptibilities of filamentous fungi.

Antifungal Agents↗

PubMed1998

In vitro activity of two echinocandin derivatives, LY303366 and MK-0991 (L-743,792), against clinical isolates of Aspergillus, Fusarium, Rhizopus, and other filamentous fungi.

LY303366 and MK-0991 (previously L-743,792) are new echinocandin derivatives with excellent broad-spectrum antifungal activity. We investigated the in vitro activity of LY303366, MK-0991, itraconazole, amnphotericin B, and 5-flucytosine against 51 clinical isolates of filamentous fungi, including Aspergillus flavus (10), A. fumigatus (12), Fusarium spp. (13), Rhizopus spp. (6), Pseudallescheria boydii (5), and one isolate each of Acremonium spp., A. niger, A terreus, Paecilomyces spp., and Trichoderma spp. In vitro susceptibility testing was performed using a microdilution broth method performed according to National Committee for Clinical Laboratory Standards guidelines. Ly303366 was two- to fourfold more active than MK-0991 against A. flavus, A. fumigatus, and Trichoderma spp. Both LY303366 and MK-0991 were considerably more active (MIC90 of 0.03-0.12 micrograms/mL) than itraconazole, amphotericin B, and 5-flucytosine against Aspergillus spp., but were less active than intraconazole and amphotericin B against Rhizopus spp. MK-0991 was more active than either LY303366 or intraconazole against Acremonium spp., Paecilomyces spp., and P. boydii. These data demonstrate promising activity of both LY303366 and MK-0991 against Aspergillus spp. and other species of filamentous fungi that are likely to be encountered clinically. Further in vitro and in vivo investigation is indicated.

Anidulafungin↗

PubMed2000

Impact of phosphate-solubilizing fungi on the yield and phosphorus-uptake by wheat and faba bean plants.

Three fungal isolates (phosphate-dissolvers), Aspergillus niger, A. fumigatus and Penicillium pinophilum were isolated from the rhizosphere of different plants grown in Ismailia and South Sinai Governorates. They effectively solubilized rock phosphate or tricalcium phosphate in Pikovskaya's liquid medium. In pot and column experiments, they significantly reduced pH and increased available phosphorus in the soil treated with either rock phosphate or superphosphate. The yield components of wheat and faba bean plants increased as a result of soil inoculation with the isolated fungi. Penicillium pinophilum was the most efficient isolate. It increased the yield of wheat grains by 28.9 and 32.8% in the soil treated with rock phosphate and superphosphate, respectively. Similarly, it increased the production of faba bean seeds by 14.7 and 29.4% with the same treatments. The uptake of phosphorus by both crops significantly increased due to inoculation of the soil with the tested fungi.

Aspergillus fumigatus↗

PubMed2000

The development of fungi as affected by pH and type of soil, in relation to the occurrence of bacteria and soil fungistatic activity.

A study was made on the effect of liming (Ca(OH)2) on the numbers of colony-forming units (CFU) and the biomass of fungi in loamy sand (ls) and a loose sandy soil (lss) during 90 days under laboratory conditions. Liming inhibited the growth of fungi more strongly in the lighter soil. Raising the pH of lss from its native 4.5 to 7.0 and 9.0 decreased mean fungal CFU numbers by 50%, and their biomass by 42% and 68%, respectively, in comparison with control unlimed samples. Also in ls with its native pH of 7.7, when alkalinised to 9.0 and 11.0 the fungal CFU numbers were smaller than in the control by 25% and 50%, respectively, and the fungal biomass decreased by 40% and 56%, respectively. Although in a parallel research alkalinisation has been shown to stimulate bacterial growth very strongly, especially in lss, the total microbial biomass (fungal + bacterial) declined by an average of 30% (pH 9.0) and 40% (pH 11.0) in limed ls, and by 35% (pH 7.0) and as much as 50% (pH 9.0) in lss, in comparison with the control.

Bacteria↗