A century later: rediscovery, culturing and phylogenetic analysis of Diploöspora rosea, a rare onygenalean hyphomycete.

Nearly 100 years after its first discovery, Diploöspora rosea was detected on biologically damaged parchment paper in Rome, Italy and isolated from house dust collected in Micronesia. The isolation of this culture permitted morphological study of colony characters, conidium and conidiophore development, and phylogenetic investigations using sequences of nuc 18S rDNA, internal transcribed spacers, and 28S rDNA. The results indicate that D. rosea is an onygenalean fungus, of uncertain taxonomic position, basal or sister to the Gymnoascaceae. Based on observations of the parchments using SEM-Energy Dispersive Spectroscopy, we speculate that the fungus occurs in archival and domestic environments subject to periodic wetting. Its ability to grow on all low water activity media used in the study, including malt extract agar amended with 60% sucrose, confirms its xerophilic nature.

Onychomycosis caused by Onychocola canadensis: the first report in Estonia and lessons to learn.

We report the first case of onychomycosis caused by Onychocola canadensis in Estonia. We believe that the number of nail infections caused by this fungus is underestimated due to the current diagnostic algorithm of non-dermatophytic onychomycosis. The need to define categories and criteria for 'proven' and 'probable' non-dermatophyte mold infections to promote more extensive studies in the future is also discussed.

Two new Tricladium species from streams in Alaska.

Two new species of aquatic hyphomycetes in the genus Tricladium are described from streams in Alaska, USA. Both species were isolated from submerged decaying sedges. Tricladium kelleri has blackish colonies and typical tricladioid conidia formed on sympodial conidiogenous cells. Tricladium alaskense has conidia with fine elements and 0-4 lateral branches; conidia are formed on sympodial conidiogenous cells. The two species are compared to other species in the genus and related genera using morphological characters and/or rDNA sequencing data (ITS and 28S). Molecular phylogenetic analysis placed both species in the Helotiales.

Pneumonia and lung infections due to emerging and unusual fungal pathogens.

Invasive mold infections affecting the lungs are increasing in incidence and diversity. Severely immunocompromised patients are particularly vulnerable to infection from unusual, normally nonpathogenic fungi that are found naturally in the environment. Certain fungi such as Scedosporium and the dematiaceous fungi also cause lung disease in hosts without overt immune compromise. The impacts of these emerging pathogens range from airway colonization to locally invasive lung, and disseminated, disease. Diagnosis requires isolation and identification of the etiologic agent by either or both phenotypic and molecular biology methods. Evidence of tissue invasion on histopathology is often required to distinguish infection from colonization. Diagnostic imaging techniques are nonspecific, and there are no reliable serological biomarkers of infection. Many rare molds and yeasts demonstrate reduced in vitro susceptibility to antifungal agents. Although amphotericin B formulations remain clinically useful for many of these infections, voriconazole and posaconazole are more effective for some of these difficult-to-treat pathogens. Surgical resection of diseased tissue and support of the host immune system are often required to optimize outcomes.

Molecular characterization and surfactant utilization of Scolecobasidium isolates from detergent-rich indoor environments.

Scolecobasidium, generally found in outdoor samples, were isolated from detergent-rich indoor environments. The isolates from bathrooms and washing machines, because of their exposure to detergents, might be genetically and biologically distinct from outdoor isolates. In this study, 11 Scolecobasidium isolates from detergent-rich indoor environments were examined to find the genetic and biological differences between the indoor and outdoor isolates. One isolate from a wall of a soap factory, showing similar conidia morphology with S. constricta, was phylogenetically distinct from the other Scolecobasidium spp. The 10 isolates from washing machines and bathrooms were identified as S. humicola, but these were classified into 2 groups that differed from the reference strain of S. humicola from leaves. All 11 isolates and the 4 reference strains of S. constricta and S. humicola grew on the medium containing sodium oleate and polyoxyethylene-(9)-lauryl ether, but the reference strains of the other Scolecobasidium spp. grew only on the medium containing sodium oleate. The results showed that S. humicola and S. constricta could utilize both surfactants generally included in soaps or synthetic detergents as nutrients. A further implication is that the genetic variation found in the S. humicola isolates from detergent-rich indoor environments can occur as a result of adaptation to such an environment.

New genus of trichomatous coelomycete on Myrcia fenzliana from the Brazilian Cerrado.

Megacoelomyces (type species: Megacoelomyces sanchezii), an ascomycete asexual morph infecting Myrcia fenzliana (Myrtaceae) from the Brazilian Cerrado, is described as a new genus in the Phaeosphaeriaceae (Pleosporales, Dothideomycetes, Ascomycota), based on multilocus phylogeny (three nuclear ribosomal DNA and two protein-coding genes) in addition to morphological (light and scanning electron microscopy) and ecological data.

Plasmodesmata between hyphal cells of Geotrichum candidum.

Electron-microscopic studies of three isolates of Geotrichum candidum from plants and one isolate from man showed the fine structure to be similar to other closely related, yeastlike fungi. Structures were observed that appeared similar to plasmodesmata present in higher plants.

Evaluation of extracts of Jatropha curcas and Moringa oleifera in culture media for selective inhibition of saprophytic fungal contaminants.

Most fungi occur in nature and utilize simple sources of carbohydrates and nitrogen for growth. Sabouraud's dextrose agar has been an ideal medium for primary isolation of fungi from clinical specimens, but for specimens from nonsterile sites or heavily contaminated ones, it has been necessary to include inhibitory substances such as antibiotics like chloramphenicol (antibacterial) and cycloheximide (antifungal). The problems we have in the our laboratory owing to frequent contamination of cultures and the delays in the procurement of cycloheximide have stimulated a search for alternatives in our local environment to enhance effective laboratory diagnoses of fungal infections. Purified extracts of the leaves and bark of Jatropha curcas and Moringa oleifera (common plants in our locality) were tested against clinical isolates of fungi at various concentrations to determine the minimum inhibitory concentration at which common fungal contaminants are inhibited, without affecting the growth of the pathogenic fungi sought for. At a concentration of 0.75 mg ml(-1) contaminants were totally inhibited by the leaf extracts. The bark extracts did not inhibit any fungus even at higher concentrations. From the results it was evident that the leaf extracts of both plants have potentials for use as inhibitory substances in culture media against contaminant fungi including Aspergillus spp., Penicillium spp., etc. J. curcas and M. oleifera are very common plants in our locality. They can be obtained at almost no cost and at any time needed. The benefits of these findings to mycology laboratories in a developing country are enormous.

Three new species of Piricaudiopsis from southern China.

Piricaudiopsis punicae sp. nov., P. rosae sp. nov. and P. rhaphidophorae sp. nov., occurring respectively on dead branches of Punica granatum, Rosa chinensis and Rhaphidophora decursiva were collected from tropical forest in southern China. They differ from the three previously described Piricaudiopsis species in conidial morphology and proliferation of the conidiogenous cell. The presence or absence of percurrent proliferation of the conidiogenous cells and conidial appendages and the height of conidia are recognized as important characters in the delimitation of species of Piricaudiopsis. A key to Piricaudiopsis species is provided.

Influence of conidial traits and leaf structure on attachment success of aquatic hyphomycetes on leaf litter.

Attachment of conidia on leaves is a critical first step in the life cycle of aquatic hyphomycetes in streams. In a first series of microcosm experiments, attachment success of three common aquatic hyphomycete species differing in conidial shape (compact, filiform and tetraradiate) was determined on two leaf species, black alder and downy oak. Fungal species identity and leaf surface structure significantly affected conidial attachment after 24 h. The lower sides of oak leaves with extensive tufts trapped 2.4-8.8 more conidia than the upper sides of oak leaves and both sides of alder leaves. In a second experiment with seven fungal species, attachment success of two species with tetraradiate conidia was much greater than that of two other tetraradiate and three compact conidia, which all had similar attachment success. The species with the largest spores was also the most successful, but this pattern was not consistent across the size range of tested conidia. These results highlight the importance of leaf surface structure, possibly conidial shape and size and additional properties of aquatic hyphomycete conidia in determining attachment success on leaves and they point to the potential role of these factors in structuring fungal communities on decomposing leaves in streams.

Molecular analyses uncover the phylogenetic placement of the lichenized hyphomycetous genus Cheiromycina.

The genus Cheiromycina is one of the few genera of lichenized hyphomycetes for which no sexual reproductive stages have been observed. The genus includes species from boreal to temperate regions of the Northern Hemisphere where it is found growing on bark or wood. Congeners in Cheiromycina are characterized by a noncorticate thallus, nearly immersed in the substrate and presenting powdery unpigmented sporodochia, and containing chlorococcoid photobionts. The relationships of members of Cheiromycina with other fungi are not known. Here we inferred the phylogenetic placement of Cheiromycina using three loci (nuSSU, nuLSU, and mtSSU) representing C. flabelliformis, the type species for the genus, C. petri, and C. reimeri. Our results revealed that the genus Cheiromycina is found within the family Malmideaceae (Lecanorales) where members formed a monophyletic clade sister to the genera Savoronala and Malmidea. This phylogenetic placement and the relationships of Cheiromycina with other lichenized hyphomycetous taxa are here discussed.

Antimicrobial activities of amino acid derivatives of monascus pigments.

Amino acid derivatives of monascus pigments were produced by fermentation, and their antimicrobial activities were determined. Thirty-nine l- and d-forms of amino acids were added as a precursor to the fermentation medium for derivation of pigments. Derivatives with L-Phe, D-Phe, L-Tyr, and D-Tyr exhibited high activities against Gram(+) and Gram(-) bacteria with MIC values of c. 4-8 microg mL(-1). The control red pigment exhibited minimal inhibitory concentration (MIC) values higher than 32 microg mL(-1). Derivatives with L-Asp, D-Asp, L-Tyr, and D-Tyr were effective against the filamentous fungi Aspergillus niger, Penicillium citrinum, and Candida albicans. Monascus derivatives of amino acids having a phenyl ring like Phe and Tyr derivatives showed high antimicrobial activities. Incubation of the l-Phe derivative with Bacillus subtilis caused cells to aggregate with formation of pellets. Easy adsorption of the L-Phe pigment derivative to the surface of Escherichia coli cells was observed via SEM and TEM. Addition of monascus pigment derivatives decreased the oxygen uptake rate of E. coli in culture. The antimicrobial activities of pigment derivatives are considered to be related to the reduced availability of oxygen for the cells adsorbed with pigment.

Steroid hydroxylations with Botryodiplodia malorum and Colletotrichum lini.

An improved procedure for the microbial hydroxylations of dehydroepiandrosterone (DHEA, 1) and 15 beta,16 beta-methylene-dehydroepiandrosterone (2) was studied using whole cells of Botryodiplodia malorum and Colletotrichum lini. C. lini catalyzed 7 alpha- and 15 alpha-hydroxylation of 1 and 7 alpha-hydroxylation of 2, while B. malorum gave 7 beta-hydroxylation of both the substrates. The stability of the enzymatic activity was higher in the presence of co-substrates (i.e., glucose or mannitol) allowing for repeated batches of the biotransformations. The yields of 7 alpha,15 alpha-dihydroxy-1 production were improved obtaining 5.8 gl(-1) (recovered product) from 7.0 gl(-1) of substrate. The structures of the hydroxylated products were assigned by a combination of two-dimensional NMR proton-proton and proton-carbon correlation techniques.

Streams in Quebec boreal and mixed-wood forests reveal a new aquatic hyphomycete species, Dwayaangam colodena sp. nov.

Foam from eight streams in boreal and mixed-wood forests in Québec were sampled in early and late fall 2002 to evaluate the biodiversity of their aquatic hyphomycete communities. Two regions were studied: 53-54 degrees N and 46-49 degrees N. A total of 54 species were identified. Twenty taxa were found only in the northern region, and four were unique to the southern region. A new aquatic hyphomycete, Dwayaangam colodena sp. nov., was found mostly in northern streams. It is described along with its taxonomic position.

Molecular systematics of Helicoma, Helicomyces and Helicosporium and their teleomorphs inferred from rDNA sequences.

Three genera of asexual, helical-spored fungi, Helicoma, Helicomyces and Helicosporium traditionally have been differentiated by the morphology of their conidia and conidiophores. In this paper we assessed their phylogenetic relationships from ribosomal sequences from ITS, 5.8S and partial LSU regions using maximum parsimony, maximum likelihood and Bayesian analysis. Forty-five isolates from the three genera were closely related and were within the teleomorphic genus Tubeufia sensu Barr (Tubeufiaceae, Ascomycota). Most of the species could be placed in one of the seven clades that each received 78% or greater bootstrap support. However none of the anamorphic genera were monophyletic and all but one of the clades contained species from more than one genus. The 15 isolates of Helicoma were scattered through the phylogeny and appeared in five of the clades. None of the four sections within the genus were monophyletic, although species from Helicoma sect. helicoma were concentrated in Clade A. The Helicosporium species also appeared in five clades. The four Helicomyces species were distributed among three clades. Most of the clades supported by sequence data lacked unifying morphological characters. Traditional characters such as the thickness of the conidial filament and whether conidiophores were conspicuous or reduced proved to be poor predictors of phylogenetic relationships. However some combinations of characters including conidium colour and the presence of lateral, tooth-like conidiogenous cells did appear to be predictive of genetic relationships.

[Collagenolytic activity in several species of deuteromycetes under various storage conditions].

The ability of deuteromycetes of the genera Penicillium, Aspergillus, and Botrytis to retain collagenolytic activity was studied after both 2 and 10 years of storage on a Czapek medium under a layer of mineral oil at 4 degrees C, as well as in silica gel granules at 20 and -60 degrees C. The enzymatic activity of several species, including Botrytis terrestris, Penicillium janthinellum, Penicillium chrysogenum, and Penicillium citrinum, was retained under both conditions of storage. Aspergillus repens retained enzymatic activity only if stored under a layer of mineral oil. The viability of conidia and the collagenolytic activity of Botrytis terrestris, P. janthinellum, P. chrysogenum, and Penicillium citrinum, maintained on silica gel for 10 years, depended on the storage temperature. The viability of the test strains improved after storage on a silica gel at -60 degrees C. A strain of Aspergillus repens lost its ability to dissolve collagen at various storage temperatures on the silica gel. The index of lysis for three strains of Penicillium deuteromycetes (Penicillium janthinellum, Penicillium chrysogenum, and Penicillium citrinum) increased after a 10-year storage on silica gel at -60 degrees C.

Diffusible and volatile compounds produced by an antagonistic Bacillus subtilis strain cause structural deformations in pathogenic fungi in vitro.

An efficient antagonistic strain of Bacillus subtilis, originally isolated from the rhizosphere of established tea bushes, was found to cause structural deformities in six pathogenic fungi under in vitro culture conditions. This effect was attributed to the production of diffusible and volatile antifungal compounds. Out of the selected test fungi four were phytopathogenic, while the remaining two were of clinical importance. The bacterial strain successfully restricted the growth of all test fungi in dual cultures, and induced morphological abnormalities such as mycelial and conidial deviations. The inhibitory effect caused by volatiles was greater than that by diffusible compounds.

Human pulmonary granulomas caused by Chrysosporium parvum var. crescens (Emmonsia crescens).

Multiple lung nodules were found at autopsy of a 56-year-old man who died of unrelated cause. The nodules contained adiaspores which were identified as those of Chrysosporium parvum var. crescens.

Acremonium recifei as an agent of mycetoma in India.

Acremonium recifei was identified as the causal agent in a case of white grain eumycotic mycetoma of the hand which occurred in South India. The identification was based on the structure of the granules in tissue and on the morphological characteristics of the culture obtained from the granules. This case represents the first documented instance in which A. recifei has been found outside of Latin America.

Phaeohyphomycotic soft tissue disease caused by Pleurophomopsis lignicola in a kidney transplant patient.

A 44-year-old immunocompromised man presented with multiple tissue abscesses, covering the entire left limb. A dematiaceous fungus compatible with Pleurophomopsis lignicola Petrak was isolated from the diseased tissue in pure culture. This is the second reported isolation of this fungus from man and the first report of fatal soft tissue infection. A detailed morphological description of the isolate is provided.