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PubMed2002

A potential resource for bioconversion of domestic wastewater sludge.

Twenty seven filamentous fungal strains representing five genera; Aspergillus, Penicillium, Trichoderma, Myriodontium and Pleurotus were isolated from four sources; domestic wastewater sludge cake (SC) from IWK (Indah Water Konsortium) wastewater treatment plant, palm oil mill effluent compost from Sri Ulu palm Oil Processing Mill, compost of plant debris, and fungal fruiting bodies from a rotten wood stump. Thirty-three strains/isolates were tested for their ability to convert domestic wastewater sludge into compost by assessing biomass production and growth rate on sludge enriched media. The strains/isolates Aspergillus niger, SS-T2008, WW-P1003 and RW-P1 512 produced the highest dry biomass at higher sludge supplemented culture media from their respective group (Aspergillus, Trichoderma, Penicillium and Basidiomycetes, respectively). This implied these strains are better adapted for growth at higher sludge rich substances, and subsequently may be efficient in bioconversion/biodegradation of sludge. The fungi isolated from ecological closely related sources were more amendable to adaptation in a sludge rich culture media.

Biodegradation, Environmental↗

PubMed1997

A quantitative bioassay for extracellular metabolites that antagonize growth of filamentous fungi, and its use with biocontrol fungi.

A bioassay and an empirically derived formula were developed to quantify fungitoxic effects. This bioassay can be easily performed and objectively read, and it is suitable for low-volume samples originating from aqueous or organic solvents. The formula defines the Inhibition Index (I), a single value that incorporates both the response to concentrations of the inhibitory compound and the persistence of inhibition. Antagonistic efficacy of metabolites produced by biocontrol strains of Trichoderma spp. were measured based on inhibition of growth of Rhizoctonia solani. Although the bioassay itself was not influenced by pH or light conditions, these factors affected metabolite production or activity. Aqueous extracts from light-grown germlings of Trichoderma virens inhibited R. solani more than extracts from germlings grown in the dark. Low pH increased the inhibitory activity of extracts from T. virens. Tests of fungal strains developed for biocontrol demonstrated that the bioassay reflected their activity both in the field and in other in vitro tests. The bioassay and formula are readily adapted for use with other fungi.

Journal Article↗

PubMed1997

Screening of fungi for the presence of the trichodiene synthase encoding sequence by hybridization to the Tri5 gene cloned from Fusarium poae.

A trichodiene synthase gene (Tri5) was amplified from F. poae by polymerase chain reaction using synthetic primers constructed on the basis of the coding portion of the same gene from F. sporotrichioides. Sequence analysis showed a high degree of similarity with other trichodiene synthase genes. A 378 bp HindIII fragment of the gene that contains the genetic information for the putative active site of the trichodiene synthase enzyme was radiolabelled and used for dot blot analysis. This probe could detect Tri5 hybridization in 1-10 ng DNA of fusaria that have the genetic potentiality to synthesize toxic trichothecene compounds, but gave no reaction with trichothecene nonproducing members of the genus. When other fungi reported to produce trichothecenes (Myrothecium, Stachybotrys, Trichoderma, Trichothecium spp.) were tested, only strains of Myrothecium and Stachybotrys gave strong positive reaction. Faint but consistent hybridization signals were obtained in four species (F. semitectum, F. tricinctum, Trichoderma viride and Trichothecium roseum) indicating the presence of nonhomologous evolutionary variants or inactive remnants of the Tri5 gene in these fungi.

Base Sequence↗

PubMed1999

Endoglucanase 28 (Cel12A), a new Phanerochaete chrysosporium cellulase.

A 28-kDa endoglucanase was isolated from the culture filtrate of Phanerochaete chrysosporium strain K3 and named EG 28. It degrades carboxymethylated cellulose and amorphous cellulose, and to a lesser degree xylan and mannan but not microcrystalline cellulose (Avicel). EG 28 is unusual among cellulases from aerobic fungi, in that it appears to lack a cellulose-binding domain and does not bind to crystalline cellulose. The enzyme is efficient at releasing short fibres from filter paper and mechanical pulp, and acts synergistically with cellobiohydrolases. Its mode of degrading filter paper appears to be different to that of endoglucanase I from Trichoderma reesei. Furthermore, EG 28 releases colour from stained cellulose beads faster than any other enzyme tested. Peptide mapping suggests that it is not a fragment of another known endoglucanases from P. chrysosporium and peptide sequences indicate that it belongs to family 12 of the glycosyl hydrolases. EG 28 is glycosylated. The biological function of the enzyme is discussed, and it is hypothesized that it is homologous to EG III in Trichoderma reesei and the role of the enzyme is to make the cellulose in wood more accessible to other cellulases.

Amino Acid Sequence↗

PubMed2003

Fungal flora of normal eyes of healthy horses from the State of Rio de Janeiro, Brazil.

The conjunctival fungal flora of 32 adult horses with normal eyes (n = 64) from the State of Rio de Janeiro in Brazil was identified in the fall of 2000 using horses of different breeds, both genders and aged 5-19 years old. The culture samples were taken from the conjunctival sac of both eyes with a sterile cotton swab wetted with saline solution, seeded in Sabouraud's dextrose agar with chloramphenicol, and incubated for 5 days at an average temperature of 25 degrees C. The number of fungal colonies per eye varied between 0 and 250 colony forming units (CFUs). There were often differences in colony types between eyes of the same animal. Filamentous fungi of genera were isolated and identified in the following proportion of the total genera of fungal colonies isolated: Aspergillus (32.2%), Penicillium (25.8%), Scopulariopsis (15.9%), Trichoderma (11.2%), Cladosporium (5.6%), Mucor (2.1%), Syncephalastrum (2.1%), Eurotium (1.7%), Geotrichum (0.9%), Rhizopus (0.9%), Gliomastix (0.4%), Fusarium (0.4%), Staphylotrichum (0.4%) and Verticillium (0.4%). Yeast genera represented 9% of the total isolates. Over half the horses had at least one normal eye with either Aspergillus, Penicillium, Trichoderma or Scopulariopsis isolated, which is a departure from other studies of the normal horse eye.

Animals↗

PubMed2003

Allergen detection from 11 fungal species before and after germination.

BACKGROUND: Allergens dispersed by airborne fungal spores play an important but poorly understood role in the underlying cause and exacerbation of asthma. Previous studies suggest that spores of Alternaria and Aspergillus release greater quantities of allergen after germination than before germination. It is unknown whether this is true of other allergenic fungi. OBJECTIVE: Our purpose was to investigate the release of allergen from a range of individual fungal spores before and after germination. METHODS: Allergen expression from spores of Alternaria alternata, Cladosporium herbarum, Aspergillus fumigatus, Botrytis cinerea, Epicoccum nigrum, Exserohilum rostratum, Penicillium chrysogenum, Stemphylium botryosum, Curvularia lunata, Trichoderma viride, and Bipolaris spicifera was examined by halogen immunoassays through the use of pooled serum IgE from patients allergic to fungus. Spores were deposited onto protein-binding membranes direct from culture. To germinate spores, samples were incubated in high humidity at room temperature for 48 hours. Ungerminated and germinated samples were then laminated with an adhesive film and immunostained by the halogen assay. The samples were examined by light microscopy, and positive counts (haloed particles) were expressed as percentages of total spores. RESULTS: For 9 of 11 species, between 5.7% and 92% of spores released allergen before germination. Spores of Penicillium and Trichoderma did not release detectable allergen. After germination, all spores that germinated had allergen elution from their hyphae. Eight of 11 species showed a significant increase (P <.05) in the percentage of spores eluting detectable allergen. Localization of allergen along the hyphae varied with species, such that some eluted allergen mainly from hyphal tips and septal junctions whereas others eluted allergen along the entire length. CONCLUSIONS: Increased elution of allergen after germination might be a common feature of many species of allergenic fungi. Although allergens from both spores and hyphae were recognized by human IgE, the extent to which human exposure occurs to allergens eluted from inhaled spores or from hyphae that germinate after deposition in the respiratory tract remains to be explored. The patterns of allergen expression might affect the clinical response to such exposure.

Administration, Inhalation↗

PubMed1998

Genes from mycoparasitic fungi as a source for improving plant resistance to fungal pathogens.

Disease resistance in transgenic plants has been improved, for the first time, by the insertion of a gene from a biocontrol fungus. The gene encoding a strongly antifungal endochitinase from the mycoparasitic fungus Trichoderma harzianum was transferred to tobacco and potato. High expression levels of the fungal gene were obtained in different plant tissues, which had no visible effect on plant growth and development. Substantial differences in endochitinase activity were detected among transformants. Selected transgenic lines were highly tolerant or completely resistant to the foliar pathogens Alternaria alternata, A. solani, Botrytis cinerea, and the soilborne pathogen Rhizoctonia solani. The high level and the broad spectrum of resistance obtained with a single chitinase gene from Trichoderma overcome the limited efficacy of transgenic expression in plants of chitinase genes from plants and bacteria. These results demonstrate a rich source of genes from biocontrol fungi that can be used to control diseases in plants.

Base Sequence↗

PubMed2002

Cleavage of the xylosyl serine linkage between a core peptide and a glycosaminoglycan chain by cellulases.

We previously found that endo-beta-xylosidase from Patinopecten is an endo-type glycosidase that cleaves the xylosyl serine linkage between a glycosaminoglycan chain and its core protein (Takagaki, K., Kon, A., Kawasaki, H., Nakamura, T., Tamura, S., and Endo, M. (1990) J. Biol. Chem. 265, 854-860). Screening for endo-beta-xylosidase activity in several cellulases detected this activity in the enzymes from Aspergillus niger, Penicillium funiculosum, Trichoderma reesei, Trichoderma viride, and Irpex lacteus. The cellulase derived from A. niger was purified, and its molecular weight was determined to be 26,000 by SDS-PAGE. Examination of the specificity of the cellulase revealed that 1) the enzyme acts on the linkage region (xylosyl serine) between a core peptide and a glycosaminoglycan chain; 2) enzymatic activity is greater with shorter glycosaminoglycan chains; 3) the enzyme readily hydrolyzes the linkage in glycosaminoglycan peptides, but intact proteoglycan is cleaved only slowly; and 4) the activity is unaffected by the glycosaminoglycan component (chondroitin sulfate, dermatan sulfate, and heparan sulfate). Judging from these enzymatic characteristics, this cellulase is different from the endo-beta-xylosidase of Patinopecten. We believe that this cellulase will become a useful tool in the further development of glycotechnology, because, like the endo-beta-xylosidase of Patinopecten, it enables the release of intact glycosaminoglycans from glycosaminoglycan peptides.

Carbohydrate Sequence↗

PubMed1999

The abfB gene encoding the major alpha-L-arabinofuranosidase of Aspergillus nidulans: nucleotide sequence, regulation and construction of a disrupted strain.

Using a DNA fragment containing the Aspergillus niger abfB gene as a probe, the homologous Aspergillus nidulans gene, designated abfB, has been cloned from a genomic library containing size-selected HindIII fragments. The nucleotide sequence of the A. nidulans abfB gene shows strong homology with the A. niger abfB, Trichoderma reesei abf-1 and Trichoderma koningii alpha-L-arabinofuranosidase/beta-xylosidase genes. Regulation of abfB expression has been investigated in cultures induced with L-arabitol. The accumulation of abfB mRNA, total alpha-L-arabinofuranosidase activity and AbfB protein levels have been determined in a wild-type A. nidulans strain as well as in different mutant strains. These strains are affected either in their response to ambient pH (paIA1 and pacC(c)14 mutants), carbon catabolite repression (creA(d)4 mutant), the ability to utilize L-arabitol as a carbon source (araA1 mutant) or a combination of both latter genotypes (araA1 creA(d)4). The results obtained indicate that the expression of the A. nidulans abfB gene was higher at acidic pHs and was superinduced in this double mutant. Furthermore, disruption of the abfB gene demonstrated that in A. nidulans AbfB is the major p-nitrophenyl alpha-L-arabinofuranoside-hydrolysing activity but at least one minor activity is expressed, which is involved in the release of L-arabinose from polysaccharides.

Amino Acid Sequence↗

PubMed2002

Endophytic fungal beta-1,6-glucanase expression in the infected host grass.

Mutualistic fungal endophytes infect many grass species and often confer benefits to the hosts such as reduced herbivory by insects and animals. The physiological interactions between the endophytes and their hosts have not been well characterized. Fungal-secreted proteins are likely to be important components of the interaction. In the interaction between Poa ampla and the endophyte Neotyphodium sp., a fungal beta-1,6-glucanase is secreted into the apoplast, and activity of the enzyme is detectable in endophyte-infected plants. Sequence analysis indicates the beta-1,6-glucanase is homologous to enzymes secreted by the mycoparasitic fungi Trichoderma harzianum and Trichoderma virens. DNA gel-blot analysis indicated the beta-1,6-glucanase was encoded by a single gene. As a secreted protein, the beta-1,6-glucanase may have a nutritional role for the fungus. In culture, beta-1,6-glucanase activity was induced in the presence of beta-1,6-glucans. From RNA gel blots, similar beta-1,6-glucanases were expressed in tall fescue (Festuca arundinacea Schreb.) and Chewings fescue (Festuca rubra L. subsp. fallax [Thuill] Nyman) infected with the endophyte species Neotyphodium coenophialum and Epichloë festucae, respectively.

Amino Acid Sequence↗

PubMed1999

Characterization of two novel type I ribosome-inactivating proteins from the storage roots of the andean crop Mirabilis expansa.

Two novel type I ribosome-inactivating proteins (RIPs) were found in the storage roots of Mirabilis expansa, an underutilized Andean root crop. The two RIPs, named ME1 and ME2, were purified to homogeneity by ammonium sulfate precipitation, cation-exchange perfusion chromatography, and C4 reverse-phase chromatography. The two proteins were found to be similar in size (27 and 27.5 kD) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their isoelectric points were determined to be greater than pH 10.0. Amino acid N-terminal sequencing revealed that both ME1 and ME2 had conserved residues characteristic of RIPs. Amino acid composition and western-blot analysis further suggested a structural similarity between ME1 and ME2. ME2 showed high similarity to the Mirabilis jalapa antiviral protein, a type I RIP. Depurination of yeast 26S rRNA by ME1 and ME2 demonstrated their ribosome-inactivating activity. Because these two proteins were isolated from roots, their antimicrobial activity was tested against root-rot microorganisms, among others. ME1 and ME2 were active against several fungi, including Pythium irregulare, Fusarium oxysporum solani, Alternaria solani, Trichoderma reesei, and Trichoderma harzianum, and an additive antifungal effect of ME1 and ME2 was observed. Antibacterial activity of both ME1 and ME2 was observed against Pseudomonas syringae, Agrobacterium tumefaciens, Agrobacterium radiobacter, and others.

Amino Acid Sequence↗

PubMed1983

Fungal biodegradation and biotransformation of soluble lignocarbohydrate complexes from straw.

Aspergillus japonicus is an efficient degrader of phenolics and carbohydrates present in a mixture of soluble lignocarbohydrate complexes extracted from wheat straw. Trichoderma sp. attacked part of the carbohydrate but hardly affected the aromatic portion of this solution. Polyporus versicolor had a complex effect; polymerization of low-molecular-size phenolics accompanied the degradation of aromatic and carbohydrate polymers. The addition of xylose to the medium facilitated depolymerization of lignin by the fungi tested and prevented the polymerization of low-molecular-size fractions of lignocarbohydrate complexes by P. versicolor. P. versicolor, in contrast to A. japonicus and Trichoderma sp., also excreted into the medium considerable amounts of laccase, but only in the absence of endogenous or exogenous carbohydrates. Apparently, laccase is involved in polymerization rather than degradation of lignin in this organism. A number of extracellular glycanases were also secreted by these fungi.

Journal Article↗

PubMed1970

Relative numbers of certain microbial groups present in compost used for mushroom (Agaricus bisporus) propagation.

The relative numbers of microorganisms associated with compost during mushroom production were studied by the dilution plate method. Thermophilic actinomycetes and fungi were isolated with a very high frequency early in the growing season. Although numbers of thermophilic bacteria diminished slowly during the season, the thermophilic fungi and actinomycetes diminished rapidly with the latter disappearing after 6 weeks. Mesophilic fungi other than Agaricus or Trichoderma remained relatively stable throughout the growing period. Agaricus could be isolated between the first and third break. Trichoderma became dominant after the fourth break. The mesophilic bacterial counts diminished during the most productive portion of the mushroom cropping season and then increased to much higher numbers toward the end of the season.

Actinomyces↗

PubMed1994

Respiratory symptoms, immunological responses, and aeroallergen concentrations at a sawmill.

After identification of a case of extrinsic allergic alveolitis due to exposure to wood dust at a sawmill, all employees at the sawmill where he worked were studied with an occupational, environmental, and symptom questionnaire, spirometry, skin prick tests, and serum specific IgG measurements. Ninety five of current and 14 of 17 ex-sawmill workers were studied. As a basis for comparison, a group of 58 workers from a nearby light engineering factory were also studied. Few women (6) were employed and they were excluded from the analysis. Workers at the sawmill were stratified into high and low exposure groups depending on their place of work. This division was supported both by their subjective assessment of the dustiness of their environment and the results of personal dust samples. There were no significant differences between the three groups in age, height, smoking habits, exposure to other causes of extrinsic allergic alveolitis, forced expiratory volume in one second, forced vital capacity, atopic state, or cutaneous reactivity to moulds. In the high exposure group the prevalence of work related cough and nasal and eye symptoms was higher than in the low exposure and comparison groups. The prevalence of work related wheeze was similar in both the high exposure and comparison groups, but was lower in the low exposure group. The prevalences of chronic bronchitis and symptomatic bronchial hyper-reactivity were similar in the high and low exposure groups but were lower in the comparison group. Serum concentrations of specific IgG against extracts of sawdust and Trichoderma koningii were significantly higher in the high exposure group than in the other two groups. The prevalence of symptoms suggestive of extrinsic allergic alveolitis was 4.4% in the high exposure group, greater than in the low exposure group (0%), and the comparison group (1.9%). In conclusion extrinsic allergic alveolitis probably occurs in British sawmills, and among the exposed population its prevalence may be as high as that reported in Sweden. The allergen responsible is likely to be from mould growing on the wood and may be from Trichoderma koningii.

Adolescent↗

PubMed1975

Effect on microorganisms of volatile compounds released from germinating seeds.

Volatile compounds evolved from germinating seeds of slash pine, bean, cabbage, corn, cucumber, and pea were evaluated for their ability to support growth of microorganisms in liquid mineral salts media lacking a carbon source. Growth of eight bacteria was measured turbidimetrically and of six fungi as dry weight of mycelium. Volatiles caused increased growth of Pseudomonas fluorescens, Bacillus cereus, Erwinia carotovora, Agrobacterium tumefaciens, A. radiobacter, Rhizobium japonicum, Mucor mucedo, Fusarium oxysporum f. conglutinans, Trichoderma viride, and Penicillium vermiculatum but not of Sarcina lutea, Serratia marcescens, Chaetomium globosum, or Schizophyllum commune. Spores of Trichoderma viride showed higher germination in the presence of volatiles. Effects on growth were apparent only during the first 3 or 4 days after planting the seeds. Killed or dried seeds had no effect. The volatiles did not support microbial growth in the absence of nitrogen nor did they supply growth factors. Passing volatiles through KMnO4 or hydrazone reduced growth of the bacteria, indicating that oxidizable organic compounds, primarily aldehydes, were the active components. The volatiles were not absorbed by sterile soil, clay minerals, or water, but they were absorbed by non-steril soil and activated charcoal.

Aldehydes↗

PubMed2001

Biological control in greenhouse systems.

The controlled environment of greenhouses, the high value of the crops, and the limited number of registered fungicides offer a unique niche for the biological control of plant diseases. During the past ten years, over 80 biocontrol products have been marketed worldwide. A large percentage of these have been developed for greenhouse crops. Products to control soilborne pathogens such as Sclerotinia, Pythium, Rhizoctonia and Fusarium include Coniothyrium minitans, species of Gliocladium, Trichoderma, Streptomyces, and Bacillus, and nonpathogenic Fusarium. Products containing Trichoderma, Ampelomyces quisqualis, Bacillus, and Ulocladium are being developed to control the primary foliar diseases, Botrytis and powdery mildew. The development of Pseudomonas for the control of Pythium diseases in hydroponics and Pseudozyma flocculosa for the control of powdery mildew by two Canadian research programs is presented. In the future, biological control of diseases in greenhouses could predominate over chemical pesticides, in the same way that biological control of greenhouse insects predominates in the United Kingdom. The limitations in formulation, registration, and commercialization are discussed, along with suggested future research priorities.

Canada↗

PubMed2005

Virulence of fungal spores determined by tracheal inoculation of goats following inhalation of aerosolized sterile feedyard dust.

OBJECTIVE: To compare the virulence of spores of 7 fungi by tracheal inoculation of goats following exposure of goats to an aerosol of sterilized feedyard dust. Animals-54 weanling Boer-Spanish goats. PROCEDURE: A prospective randomized controlled study was conducted. There were 7 fungal treatment groups, a tent control group, and a pen control group (n = 6 goats/group). Goats in the 7 treatment and tent control groups were exposed to autoclaved aerosolized feedyard dust for 4 hours in a specially constructed tent. Goats in the 7 treatment groups were then inoculated intratracheally with 30 mL of a fungal spore preparation, whereas tent control goats were intratracheally inoculated with 30 mL of physiologic saline (0.9% NaCI) solution. These treatments were repeated each week for 6 weeks. RESULTS: Severity of pathologic changes differed significantly among the 7 fungal treatment groups as determined on the basis of gross atelectatic and consolidated lung lesions and histologic lesions of the lungs. Descending order for severity of lesions was Mucor ramosissimus, Trichoderma viride, Chaetomium globosum, Stachybotrys chartarum, Aspergillus fumigatus, Penicillium chrysogenum, and Monotospora lanuginosa. Trichoderma viride spores were the most invasive and were isolated from the bronchial lymph nodes and thoracic fluid of all 6 goats administered this organism. Spores were observed-histologically in lung tissues harvested 72 hours after inoculation from all treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: 4 of 7 fungal spore types induced significantly larger lung lesions, compared with those induced by the other 3 spore types or those evident in control goats.

Air Microbiology↗

PubMedDate not supplied

Hypocrea voglmayrii sp. nov. from the Austrian Alps represents a new phylogenetic clade in Hypocrea/Trichoderma.

The holomorph of the new species Hypocrea voglmayrii (Hypocreales, Ascomycota, Fungi) is described by a combined approach, using morphology of the teleomorph, morphology of the anamorph, culture studies and phylogenetic analyses of ITS1 and 2, ech42 and rpb2 gene sequences. Its anamorph Trichoderma voglmayrii is described as a new anamorph species. Unlike most other species of Hypocrea the teleomorph of H. voglmayrii occurs on dry standing trunks and exhibits well defined black ostioles. Although exclusively collected at higher altitudes, this species grows at 35 C in culture. Hypocrea voglmayrii develops pale yellowish to greenish conidia, a yellowish pigment and a coconut-like odor on CMD. Phylogenetically, H. voglmayrii forms a distinct, isolated branch between the section Trichoderma and the H. pachybasioides clade but does not associate with any of these clades in different gene trees.

Austria↗