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Antibacterial activity of marine-derived fungi.

A total of 227 marine isolates of ubiquitous fungi were cultivated on different media and the secondary metabolite content of the extracts (ethyl acetate/chloroform/methanol 3:2:1) characterized by HPLC. The fungi were secured from animals, plants and sediments of Venezuelan waters (0-10 m) including mangroves and lagoonal areas. The extracts were tested for antibacterial activity. A total of 7 were active towards Vibrio parahaemolyticus and 55 towards Staphylococcus aureus, representing 18 different fungal species from 8 ascomycetous genera. For 61 strains of Penicillium citrinum antibacterial activity correlated well with content of secondary metabolites as measured by HPLC. Thirteen isolates of Penicillium steckii produced very similar profiles of secondary metabolites and 6 of these had activity against either V. parahaemolyticus or S. aureus or both.

Anti-Bacterial Agents↗

Effect of cadmium on growth of potentially pathogenic soil fungi.

The effect of Cd on the mycelial growth of some potentially pathogenic soil fungi was investigated. Sixty-four strains from twenty-five fungal species were tested for their susceptibility to Cd. Final colony diameter, radial growth rate and final dry mass of mycelium (for Cd: 1-200 ppm) were measured. EcD50 values were calculated from these parameters. The intra- and interspecific variability in the results obtained is presented. Among the keratinolytic fungal group, the general Arthrographis, trichophyton and Chrysosporium (including Chrysosporium anamorph of Aphanoascus reticulisporus) were the most resistant to Cd. T. mentagrophytes was the most sensitive of all Trichophyton species tested. Among the nonkeratinolytic fungal group, the genera Pseudallescheria, Absida and Rhizopus were highly resistant to Cd. Strains of Aspergillus were more resistant to Cd than strains of Penicillium. Zygomycetes were more resistant to Cd than Ascomycetes with Fungi Imperfecti. Nonkeratinolytic fungi showed higher resistance to Cd than keratinolytic fungi. The two last differences resulted from the extremely high EcD50 values for R. oryzae and A. corymbifera.

Analysis of Variance↗

Pathogenicity of Hansenula anomala in model of immunocompromised mice.

Systemic infections caused by opportunistic fungi have shown an increased frequency in the past 10 years, particularly in immunocompromised patients. Hansenula anomala is an ascosporogenous yeast of the Ascomycetes class found in the skin, throat, and digestive tract transient normal flora. This study was conducted to compare the pathogenicity of H. anomala and Candida albicans in a model of immunocompromised mice. Thirty-eight Swiss mice were divided into two groups as follows: 30 animals received an intraperitoneal (i.p.) injection of cyclophosphamide (200 mg/kg) four days before the induction of infection with H. anomala (1 x 10(6) yeasts/mL), and 8 animals received 100 mg/kg of cyclophosphamide at 3-day intervals during 3 weeks before inoculation of 1 x 10(7) yeasts/mL. All animals were treated with amoxicillin/clavulanic acid (40 mg/kg) four days before induction of infection. A group of mice inoculated with C. albicans (ATCC 64548) served as control. Tissue samples from the lung, spleen, liver, and kidney for histological and mycologic studies were obtained at necropsy. In each animal, the number of viable yeasts per gram of kidney was determined. The organs most frequently infected by H. anomala were the kidneys and the liver (20%), and the lung (10%). However, in conditions of sustained immunosuppression, H. anomala was found in 65.5% of the organs examined. It is concluded that in an experimental model of immunocompromised mice, the pathogenicity of H. anomala was low.

Amoxicillin↗

Degradation of polycyclic aromatic hydrocarbons with three to seven aromatic rings by higher fungi in sterile and unsterile soils.

Seven commercial 3- to 7-ring (R) polycyclic aromatic hydrocarbons (PAH) as well as PAH derived from lignite tar were spiked into 3 soils (0.8 to 9.7% of organic carbon). The disappearance of the original PAH was determined for the freshly spiked soils, for soils incubated for up to 287 d with their indigenous microflora, and for autoclaved, unsterile and pasteurized soils inoculated with basidiomycetous and ascomycetous fungi. Three to 12 d after spiking, 22 to 38% of the PAH could no longer be recovered from the soils. At 287 d, 88.5 to 92.7%, 83.4 to 87.4%, and 22.0 to 42.1% of the 3-, 4-, and 5- to 7-R PAH, respectively, had disappeared from the unsterile, uninoculated soils. In 2 organic-rich sterile soils, the groups of wood- and straw-degrading, terricolous, and ectomycorrhizal fungi reduced the concentration of 5 PAH by 12.6, 37.9, and 9.4% in 287 d. Five- to 7-R PAH were degraded as efficiently as most of the 3- to 4-R PAH. In organic-rich unsterile soils inoculated with wood- and straw-degrading fungi, the degradation of 3- to 4-R PAH was not accelerated by the presence of fungi. The 5- to 7-R PAH, which were not attacked by bacteria, were degraded by fungi to 29 to 42% in optimum combinations of fungal species and soil type. In organic-poor unsterile soil, these same fungi delayed the net degradation of PAH possibly for 2 reasons. Mycelia of Pleurotus killed most of the indigenous soil bacteria expected to take part in the degradation of PAH, whereas those of Hypholoma and Stropharia promoted the development of opportunistic bacteria in the soil, which must not necessarily be PAH degraders. Contemporarily, the contribution of the fungi themselves to PAH degradation may be negligible in the absence of soil organic matter due to the lower production of ligninolytic enzymes. It is concluded that fungi degrade PAH irrespective of their molecular size in organic-rich and wood chip-amended soils which promote fungal oxidative enzyme production.

Biodegradation, Environmental↗

A review of soybean (Glycine max) seed, pod, and flower mycofloras in North America, with methods and a key for identification of selected fungi.

A review of the fungi associated with soybean seeds, pods, or flowers was conducted in North America. Species of Deuteromycetes are the most common fungi in each of the soybean flower organs followed by the Ascomycetes and Phycomycetes which comprise about one-fourth of the total mycoflora. Eighty genera and about 135 or more species occur in seeds, pods, or flowers. With regard to numbers of taxa from separate mycofloras, 63 genera and about 108 or more species occur in seeds, 65 genera and about 88 or more species occur in pods, and 36 genera and approximately 47 or more species occur in flowers. Most of the fungi which occur in flowers can be cultured from pods, and the majority of those fungi occur in seeds. Methods for and a key are provided for the identification of the 30 most important selected fungi.

Ascomycota↗

Fungal populations on sunflower (Helianthus annuus) anthosphere and their relation to susceptibility or tolerance to Sclerotinia sclerotiorum attack.

Analysis of the fungal flora from different floret parts of various sunflower (Helianthus annuus) varieties showed that there are differences in both fungal species and frequency, depending on whether the sunflower variety is susceptible (SV) or tolerant (TV) to attack of the flower heads by the ascomycete pathogen Sclerotinia sclerotiorum. The sunflower varieties analyzed were SV: HA 300 and Z 20028, and TV: HA 302, Z AV 8410 and Z 30629. The isolates showed different "in vitro" behavior as biocontrol agents. The most common types of interaction with Sclerotinia sclerotiorum were D2 and D2+ (hyphal contact) for isolates from SV and TV, while some of the isolates from TV displayed antibiosis. The microorganisms that colonize TV florets play a part in an indirect mechanism that protects flowers from ascospore germination and pathogen growth.

Ascomycota↗

Two new anamorphic yeasts: Candida germanica and Candida neerlandica.

Descriptions are given for the two new anamorphic ascomycetous yeasts Candida germanica (type strain NRRL Y-27064, CBS 4105) and Candida neerlandica (type strain NRRL Y-27057, CBS 434). The species were isolated, respectively, from the atmosphere over Germany and from pressed yeast cake in The Netherlands. Phylogenetic analysis of 26S domain D1/D2 ribosomal DNA sequences places C. germanica near Pichia philogaea, whereas C. neerlandica is a member of the Lodderomyces elongisporus/Candida albicans clade.

Air Microbiology↗

Three new species of Candida from apple cider: C. anglica, C. cidri and C. pomicola.

Three new anamorphic ascomycetous yeasts are described: Candida anglica (type strain NRRL Y-27079, CBS 4262), Candida cidri (type strain NRRL Y-27078, CBS 4241), and Candida pomicola (type strain NRRL Y-27083, CBS 4242). These three species were isolated from cider produced in the United Kingdom, and their identification was determined from unique nucleotide sequences in the species-specific D1/D2 domain of large subunit (26S) ribosomal DNA. Phylogenetic analysis of D1/D2 sequences placed C. anglica near Candida fragi, C. cidri near Pichia capsulata, and C. pomicola in the Pichia holstii clade.

Beverages↗

Ferricrocin--an ectomycorrhizal siderophore of Cenococcum geophilum.

The ectomycorrhizal fungus Cenococcum geophilum was grown in low-iron medium and the excreted siderophores were extracted, purified and analyzed by HPLC. The principal hydroxamate siderophore produced, was identified as ferricrocin as confirmed by analytical HPLC, FAB-mass spectrometry and 1H- and 13C-NMR spectra. Although the occurrence of ferricrocin has been shown earlier to occur in the ericoid mycorrhizal fungi, this is the first report of ferricrocin in a true ectomycorrhizal fungus which is taxonomically related to the ascomycetes.

Ascomycota↗

A research on the microfungal flora of some greenhouse soils in the vicinity of Lapseki Canakkale, Turkey.

Qualitative and quantitative distributions of microfungal flora of the some greenhouse soils in the vicinity of Lapseki, Canakkale were studied. A total of 25 soil samples were collected from 10 cm depth in five greenhouses. Using soil dilution technique, 128 different microfungal isolates were obtained Isolates collected belonged to Oomycetes (1), Zygomycetes (7), Ascomycetes (9) and 96 belonging to the Deuteromycetes Fifteen isolates were classified as Mycelia Sterilia. The genera of microfungi most abundant in the greenhouse soils were Aspergillus, Penicillium, Geomyces, Exophiala and Fusarium. Qualitative and quantitative distributions of the Aspergillus were higher than those of the other genera. It was found that the maximum value was in greenhouse 5 and the minimum value was in greenhouse 4. The solarization method was the most effective in reducing fungal numbers.

Ascomycota↗

Plant cell membranes as biochemical targets of the phytotoxin helminthosporol.

Helminthosporol is one of the natural sesquiterpenoid toxins isolated and identified in the culture medium of the phytopathogenic ascomycete fungus Cochliobolus sativus. The effect of this phytotoxin was investigated on enzymatic activities, electron and ion transport in mitochondria, chloroplasts, and microsomes of plant. The results indicate that helminthosporol drastically affects the membrane permeability of these organelles to protons and substrate anions, inhibiting the mitochondrial oxidative phosphorylation, the photophosphorylation in chloroplasts, and the proton pumping across the cell plasma membrane. The 1,3-beta-glucan synthase activity, involved in defense mechanisms of plant cells against stress and damage, e.g., during pathogen attack, was also strongly inhibited by the toxin.

Cell Membrane↗

A new species and three new combinations in Cyphellophora, with a note on the taxonomic affinities of the genus, and its relation to Kumbhamaya and Pseudomicrodochium.

Cyphellophora guyanensis sp. nov., isolated from leaf litter in French Guyana, is described and illustrated. This fungus is characterized by ampulliform to flask-shaped phialides, with a conspicuous, funnel-shaped collarette, and nearly straight to falcate or slightly sigmoid, (2-)3-6 septate conidia. Cyphellophora guyanensis is compared to the other Cyphellophora species, and the latter genus is compared to Kumbhamaya and Pseudomicrodochium. Kumbhamaya indica, P. fusarioides, and P. suttonii are transferred to Cyphellophora. The phylogenetic relationships of Cyphellophora with other genera of ascomycetes and related mitosporic fungi were inferred from the analysis of the partial nuclear ribosomal DNA SSU sequences data. The analysis supports a placement of Cyphellophora in Chaetothyriales, and probably in the Herpotrichiellaceae

Ascomycota↗

Assessment of yeast diversity in a marine environment in the south of Portugal by microsatellite-primed PCR.

The occurrence and diversity of yeasts in seawater was investigated in a study site located 20 Km off Faro, Portugal, above the Alvares Cabral Trench. A total of 43 water samples from different layers (above the permanent thermocline, under the thermocline and near the bottom) and directly from the surface, originated 234 isolates. All the isolates were identified using a molecular approach that included, in a first stage, MSP-PCR fingerprinting. A total of 31 MSP-PCR classes were formed, 8 for the pigmented yeasts and 23 for the non-pigmented yeasts. The pink coloured isolates were identified by direct comparison of the new fingerprints with those obtained for representative strains of the various species. For identification of the non-pigmented yeasts, a representative isolate of each MSP-PCR class was selected for sequence analysis and compared with reference sequences. The five most abundant yeast species were Sakaguchia dacryoidea, Pseudozyma aphidis, Rhodosporidium babjevae, R. diobovatum and Debaryomyces hansenii. The distribution of isolates and species in the major taxonomic groups indicated that the number of basidiomycetous yeasts and their diversity are prevalent in relation to their ascomycetous counterpart. Diversity indices were determined and superficial water and water near the bottom had the highest diversity. The sampling effort effectiveness was estimated, and found to correspond to approximately 60% of the species present. MSP-PCR identification proved suitable for pigmented basidiomycetous yeasts and, when used in conjunction with sequence analysis, was effective for the characterization of non-pigmented populations. Our results indicate that the MSP-PCR fingerprinting method is appropriate for the characterization of large groups of isolates due to its simplicity and good reproducibility.

Biodiversity↗

Screening of basidiomycetes for antimicrobial activities.

As a part of a screening programme developed to evaluate the antimicrobial activity of basidiomycetes, 317 isolates representing 204 species collected in Spain were screened against a range of human clinical pathogens and laboratory controls. Extracts from 45% of the isolates, representing 109 species, showed antimicrobial activity. Antibacterial activity was more pronounced than antifungal activity. The proportion of extracts from basidiomycetes showing antimicrobial activity was similar to or above that obtained for representative orders of Ascomycetes, such as Pezizales and Xylariales, but lower than that produced by members of the orders Diaporthales, Eurotiales, Hypocreales, Leotiales and Sordariales. Suprageneric taxa (orders and families) did not show pronounced differences in their antimicrobial activities though such differences were observed at the genus level, suggesting that the ability to produce these bioactive compounds is not homogenously distributed amongst the basidiomycetes. Isolates from some species showed large differences in their ability to produce metabolites with antimicrobial activity, possibly reflecting genetic differences at the infraspecific level.

Anti-Bacterial Agents↗

Homokaryotic and heterokaryotic hyphae in Terfezia.

Mycelia of Terfezia pfeilii (Ascomycetes) were obtained by two methods, i.e., from the sterile hyphae of fresh fruit bodies or by germinating ascospores. Nuclear staining revealed the existence of multinucleate cells in all mycelia. Paired nuclei were observed only in mycelia obtained from sterile hyphae proliferation, while single nuclei were found in mycelia originating from singly germinated spores. Co-cultivation of mycelia from two different ascospores apparently facilitated plasmogamy, resulting in mycelia with paired nuclei. Terfezia boudieri cultures originating from sterile hyphae also exhibit paired nuclei, indicating the possible existence of a long-term heterokaryon. The timing of plasmogamy and karyogamy in Terfezia is discussed.

Ascomycota↗

Candida sergipensis, a new asexual yeast species isolated from frozen pulps of tropical fruits.

Sixteen strains of the new yeast species Candida sergipensis have been isolated from frozen pulps of the tropical fruits umbú ( Spondias tuberosa Avr. Cam.) and mangaba ( Hancornia speciosa Gom.). Candida sergipensis was one of the prevalent species in the yeast community of these substrates. The new asexual ascomycetous yeast is phylogenetically related to Candida spandovensis and Candida sorbophila, species belonging to the Wickerhamiella clade, as evidenced by the sequences of the D1/D2 domains of their large subunit ribosomal DNAs. The species C. sergipensis and C. spandovensis can be separated on the basis of growth on 50% glucose agar, xylose and succinate, negative for the first species and positive for the second. The type culture is strain UFMG-R188 (CBS 9567).

Anacardiaceae↗

Pod and seed mycoflora on transgenic and conventional soybean [Glycine max (L.) Merrill] cultivars in Mississippi.

A 2-year (1999-2000) study was conducted at Starkville and Stoneville, MS to determine if the occurrence of the mycoflora varied on Roundup Ready (transgenic) compared to conventional soybean (Glycine max) cultivars. A total of 7,658 fungal isolates were identified from the pod and seed tissues of four cultivars compared at growth stages R6 and R8. Ninety-nine percent of all fungi isolated were mitosporic fungi and ascomycetes. In both years, total fungal isolates from the two locations were greater from the pod (65%) than from seed (33%) tissues. Isolation frequency from conventional cultivars was 54% compared to 46% for the transgenic cultivars. The most common fungi identified that are reported pathogens of soybean included Alternaria, Cercospora, Cladosporium, Diaporthe, Fusarium and Verticillium spp. When main effects and interactions were compared among the frequency data for the fungal genera, significant differences occurred, but consistent trends were not noted. Isolation frequencies of Diaporthe spp. during the R6 growth stage, were significantly greater on the conventional than on the transgenic cultivars in both years of the study, but only at Starkville. Isolation frequencies from samples taken during the R8 growth stage were similar at both locations in 1999 and 2000. Fusarium spp. isolated at R6 and R8 growth stages from pod and seed tissues were significantly greater on conventional than on transgenic cultivars in 2000. Even though frequencies were often significantly different between the transgenic and conventional cultivars, the data was not consistent between locations, pod and seed tissues, or growth stages. The pod and seed mycoflora of transgenic and conventional soybean cultivars was, therefore, similar in Mississippi.

Antifungal Agents↗

Entomopathogenic fungi from 'El Eden' Ecological Reserve, Quintana Roo, Mexico.

Entomopathogenic fungi were isolated and identified from insects collected from the tropical forest and an agricultural area at El Eden Ecological Reserve, Quintana Roo, Mexico. These fungi were studied to determine their potential as biological control agents of greenhouse Trialeurodes vaporariorum (Homoptera: Aleyrodidae), and to contribute to the knowledge of biodiversity of this area. No pest insects were observed in the tropical forest. In contrast, all insects collected in the agricultural area were considered important pests by the local farmers, with the whitefly, as the most relevant, plentiful in Cucurbitaceae plants. From approximately 3400 collected insects in three different surveys, different anamorphic Ascomycetes were recovered. One isolate of Aspergillus sp., two of Penicillium sp., three of Paecilomyces marquandii, and three of Verticillium sp. out of 308 insects (2.9%) from three insect orders, Hymenoptera, Diptera and Isoptera in the tropical forest. In contrast, a higher number of fungal isolates were recovered from the agricultural area: three isolates from Aspergillus parasiticus, 100 of Fusarium moniliforme, one of Aschersonia sp., and 246 of Fusarium oxysporum out of 3100 insects (11.3%) from three insect orders, Homoptera, Coleoptera and Lepidoptera. The results of this study show Fusarium moniliforme and F oxysporum as highly virulent to infected insects in the agricultural area, with 100 and 246 isolates respectively, out of 350 infected insects of 3100 studied specimens. Laboratory whitefly nymph bioassays with isolates Ed29a of F. moniliforme, Ed322 of F. oxysporum, and Ed22 of P marquandii showed 96 to 97.5% insect mortality with no significant differences (P < 0.05) among them. F. oxysporum Ed322 produced no mortality when inoculated on tomato, bean, squash and maize seedlings (with and without injuries) compared to the 100% mortality caused by phytopathogenic strains, F. oxysporum f. sp. lycopersici and F. oxysporum f. sp. radicis lycopersici.

Animals↗