First Report of <Emphasis Type="Italic">Tritirachium oryzae</Emphasis> Infection of Human Scalp

The natural habitat of Tritirachium oryzae is soil and decaying plant material. It is also an insect pathogen. As a human pathogen, it has been reported as a cause of corneal ulcers and otomycosis. The case of a 4-year-old infant is reported with Tritirachium oryzae infection of the scalp. Diagnosis was established by direct mycological study and culture that showed Tritirachium oryzae as the only agent in a pure culture. The topical treatment involved an antifungal medication to a complete cure. We report the first case of scalp dermatomycosis due to Tritirachium oryzae infection, illustrating a novel clinical manifestation.     

Taxonomy and morphology of dematiaceous fungi isolated from nature

The Hughes system of classification was applied to the identification of dematiaceous fungi isolated from nature. The majority of the isolates encountered could be identified with this system using brightfield microscopy. However, phase contrast microscopy was needed to determine the mode of conidial ontogeny with some species of Phialophora, Exophiala and Wangiella. The identifications based upon phase contrast microscopy were confirmed using the scanning electron microscope (SEM). Scanning electron photomicrographs are presented for Bispora betulina, Cladosporium trichoides, Exophiala jeanselmei, Phialophora revens, P. verrucosa and Wangiella dermatitidis. SEM observations of W. dermatitidis support the present classification of this organism as proposed by other workers. It is suggested that the Hughes system of classification and phase contrast microscopy be used to facilitate identification of isolates of dematiaceous fungi encountered in the clinical laboratory.     

Mitochondrial DNA analysis of Exophiala jeanselmei and Exophiala dermatitidis

Mitochondrial DNA (mtDNA) analysis with Hae III, Hind. III and Msp I was performed in 45 Exophiala jeanselmei strains (30 Phialophora jeanselmei and 15 Phialophora gougerotii strains) and 31 Exophiala dermatitidis strains. The results were as follows, 1) P. jeanselmei and P. gougerotii are identical, 2) E. jeanselmei is classified into 18 types based on restriction profiles, 3) two strains of E. jeanselmei CBS 577.76 and CBS 578.76 are identified as E. dermatitidis, 4) E. dermatitidis has no intraspecific variation and is definitely distinct from E. jeanselmei, 5) E. jeanselmei is suggested to be a complex organism because of extensive mtDNA polymorphism.     

Pathpgenicity of Exophiala jeanselmei for ddY mice

The pathogenicity of three cultures isolated as Phialophora jeanselmei was compared with that of three cultures of Phialophora gougerotii using ddY mice.One hundred and twenty mice were used. They were divided into 6 groups consisting of 20 each. Each culture was evaluated in 20 mice. Mice were inoculated intravenously with 0.2 ml of a 1% (wet weight/vol.) yeast-like cell suspension and sacrificed at adequate intervals until the 30th day.As results, 1) the virulence of the three cultures each of P. jeanselmei and P. gougerotii to ddY mice was mild. 2) These cultures were not neurotropic. 3) P. gougerotii survived longer in the mice than P. jeanselmei. 4) There were no major differences in histopathology of the lesions in the mice inoculated with the two taxa.     

Dematiaceous fungal pathogens isolated from nature

This study was conducted to demonstrate the presence of pathogenic dematiaceous fungi in nature. Using hamster and mouse inoculation techniques, 43 isolates of dematiaceous fungi were recovered from 39 samples of woody plant material and soil from the Virginia environment. Thirteen species were identified and included 4 Phialophora spp., 3 Cladosporium spp., 2 Exophiala spp., Sporothrix sp., Wangiella dermatitidis, Bispora betulina, and Scytalidium lignicola. Evidence is presented for the first isolations of C. trichoides from nature in the United States; these isolates proved to be pathogenic for mice in which they produced disease and death in a course similar to that seen in man. Natural isolates of Phialophora verrucosa, Phialophora repens, Exophiala jeanselmei, and Wangiella dermatitidis were identical to those species isolated from man using the following criteria: morphology, 12% gelatin reaction, and survival in laboratory animals.     

Water use and phosphorus and potassium status of wheat seedlings colonized byGaeumannomyces graminis orPhialophora radicicola

Summary The water consumption and levels of phosphorus, potassium, and total minerals were measured for wheat seedlings colonized byGaeumannomyces graminis var.tritici, Phialophora radicicola var.radicicola, orPhialophora radicicola var.graminicola. Infection byG. graminis resulted in a considerable reduction in water consumption, and reduced level of phosphorus when the supply of phosphorus to the seedlings was plentiful. Colonization byP. radicicola var.radicicola increased levels of phosphorus and potassium, but these increases varied according to the isolate of the fungus and the supply of phosphorus and potassium available to the seedlings. Colonization byP. radicicola var.graminicola resulted in reduced water consumption by the seedlings.The results are discussed in relation to stelar cell wall thickening in wheat roots colonized byP. radicicola, and the effects on nutrient uptake of mycorrhizal root systems.     

Mycoflora of the Finnish “sauna” (bath-house)

The investigation concerned the mycoflora of the old, in some cases already discarded, bath houses (sauna) situated in various parts of Finland for the purpose of finding the fungus,Phialophora pedrosoi, which causes chromoblastomycosis in man. The material, 220 specimens, consisting of thin shavings from the wooden floors and platforms, was collected from 78 bath houses. P. pedrosoi was not found in this investigation; the followingPhialophora-species:P. fastigiata, P. lagerbergii, P. lignicola andP. verrucosa were isolated, however.In total 98 various fungus species were isolated, the number of isolates was 1052. Phycomycetes represented 9.2 % of the species and 15.2 % of the isolates, Ascomycetes 4.1 % and 3.4 %, Basidiomycetes 4.1 % and 0.7 % and Fungi imperfecti 82.6 % and 80.7 % respectively of the species and the isolates. The species most frequently isolated from specimens were:Trichoderma viride, Mucor mucedo, Stemphylium consortiale, Rhinocladiella atrovirens, Arthrobotrys superba andPhialophora verrucosa.In all, 13 nematode-destroying fungus species were isolated belonging to the genera:Nematoctonus, Cephalosporium, Harposporium, Dactylaria, Arthrobotrys andDactylella. 60 % of the bath houses were infected by nematodes. Gliodendron gen. nov. with the speciesGliodendron balnicolum sp. nov. is described and illustrated.

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Zusammenfassung Die Untersuchung betraf die Mykoflora von alten, in manchen Fällen bereits aufgegebenen Badehäusern, die sich in verschiedenen Teilen Finnlands befanden, um den Pilz,Phialophora pedrosoi, den Erreger der Chromoblastomykose, zu finden. Das Material bestand aus 220 Einzelstücken, Spänen von Holzfußböden, die in 78 Badehäusern gesammelt wurden,Phialophora pedrosoi wurde in diesen Untersuchungen nicht gefunden. Die folgendenPhialophora-Arten sind isoliert worden:P. fastigiata, P. lagerbergii, P. lignicola, undP. verrucosa. Insgesamt sind 98 verschiedene Pilzarten isoliert worden. Die Anzahl der Isolierungen war 1052. Phykomyceten stellten 9,2 % von den Arten und 15,2 % der Isolierungen dar: Askomyceten 4,1 % und 3,4 %; Basidiomyceten 4,1 % und 0,7 %; Fungi imperfecti 82,6 % und 80,7 %. Die häufigst isolierten Arten waren:Trichoderma viride, Mucor mucedo, Stemphylium consortiale, Rhinocladiella atrovirens, Arthrobotrys superba undPhialophora verrucosa. Alles in allem wurden 13 Nematoden zerstörende Arten isoliert, die zu den folgenden Gattungen gehörten:Nematoctonus, Cephalosporium, Harposporium, Dactylaria, Arthrobotrys undDactylella. Sechzig Prozent der Badehäuser waren von Nematoden infektiert.Gliodendron gen. nov. ist mit der ArtGliodendron balnicolum, sp. nov. beschrieben und abgebildet worden.

     

Phialophora cyanescens sp. nov. with Phaeosclera-like synanamorph,causing white-grain mycetoma in man

The human-pathogenic fungus, Phialophora cyanescens sp. nov. is characterized by the abundant production of chlamydospore-like cell aggregates resembling the form-genus Phaeosclera Sigler et al., and by elongate phialides with ellipsoidal conidia. A brown pigment and a blue, acid-labile pigment may be present.     

habitat de Nocardia asteroides,Phialophora pedrosoi y Cryptococcus neoformans en Venezuela

Resumen Se estudiaron 336 muestras de tierra, madera y plantas provenientes de diferentes zonas del Edo. Mérida/Venezuela y Estados vecinos. Fueron inoculados cobayos y hamsters intratesticularmente y ratones por via endovenosa. Testículos y pulmones respectivamente fueron examinados por medio de cultivos e histológicamente.De 11 muestras se aislaronNocardia asteroides, de 6Phialophora pedrosoi y de unaCryptococcus neoformans.Se discuten las probables causas de porqué no se logró aislarParacoccidioides brasiliensis.

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Summary 336 samples of soil, wood and plants from several aereas of the State of Mérida/Venezuela and adjacent regions were studied.The samples were injected into the testes of guinea pigs and into mice intravenously. The testes and lungs of the animals respectively were cultured and examined histologically. From eleven samples were isolatedNocardia asteroides, from sixPhialophora pedrosoi and from oneCryptococcus neoformans.It is discussed, why it was not possible to isolateParacoccidioides brasiliensis.

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Zusammenfassung Es wurden 336 Erd-, Holz- und Pflanzenproben aus verschiedenen Gebieten des Staates Mérida/Venezuela und benachbarten Zonen untersucht. Das Material wurde Meerschweinchen intratestikulär und weissen Mäusen intravenös injiziert. Hoden und Lungen der Tiere wurden auf Nährböden verbracht und histologisch untersucht.Elfmal wurdeNocardia asteroides, sechsmalPhialophora pedrosoi und einmalCryptococcus neoformans isoliert und histologisch nachgewiesen. Es werden die möglichen Gründe diskutiert, warum es nicht gelangParacoccidioides brasiliensis zu isolieren.

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Del Instituto de Anatomía Patológica y Laboratorio de Micología de la Universidad de Los Andes, Mérida/Venezuela y del Laboratorio de Micología, Escuela de Medicina de la Universidad de Cincinnati/Ohio EEUU.

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Este trabajo pudo realizarse con la ayuda económica proporcionada por el Sr. Rector de la U.L.A., Dr.P. Rincón Gutiérrez a quien damos nuestras gracias más sinceras.A Wm. Bridge Cooke, Ph. D. y al Dr.G. A. de Vries agradecemos la identificación de las cepas dePhialophora pedrosoi.

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A los Dres.R. Kohler yP. N. Tablante G., y al Sr.Oswaldo Juergenson nuestro agradecimiento por la ayuda prestada en la recolección de las muestras.     

Isolation of Phialophora verrucosa and Fonsecaea pedrosoi from nature in Japan

Seventeen strains of Phialophora verrucosa and one strain of Fonsecaea pedrosoi were isolated from natural sources such as rotting wood, pine bark, pine logs or soil in Japan. These saprophytic isolates were compared morphologically, biologically and serologically with human isolates and their virulence for rats was studied. There were no distinct differences between the isolates from the two sources.     

Genetic Diversity of Naturally Established Ectomycorrhizal Fungi on Norway Spruce Seedlings under Nursery Conditions

We have assessed ectomycorrhizal fungi colonizing Norway spruce (Picea abies L.) seedlings in nine forest nurseries using restriction fragment length polymorphism (RFLP) and sequencing analyses of the internal transcribed spacers (ITS1-5.8S-ITS2) amplicons. Restriction analysis of the amplified DNA fragments with HinfI, MboI, and TaqI enzymes allowed the definition of 17 RFLP genotypes; five of them could be unambiguously assigned to Thelephora terrestris, Hebeloma longicaudum, H. crustuliniforme, Tricharina ochroleuca, and Cenococcum geophilum species by comparison with the sporocarp RFLP-pattern database. The remaining genotypes have been sequenced and compared with sequences deposited in the GenBank database. The phylogenetic analysis of resulting sequences and their identified matches indicated that isolated genotypes have formed seven clades. The ascomycetes were predominant: we have determined eight species—Wilcoxina mikolae, Phialophora finlandia, Tuber sp., Cenococcum geophilum, Tricharina ochroleuca, Pulvinula constellatio, and two unidentified ascomycetes—whereas the basidiomycetes were less common (four species denoted: Amphinema byssoides, Hebeloma crustuliniforme, H. longicaudum, and Thelephora terrestris). Wilcoxina mikolae and Phialophora finlandia were the most frequent fungi. Analysis of variance revealed that ascomycetes abundance was higher in nurseries that used organic fertilizer.     

Characterization of the nuclear DNA ofPhialophora gregata ff.sp.adzukicola andsojae

Phialophora gregata nuclear (n) DNA was characterized by physical methods. The nDNA of f.sp.adzukicola was shown to be larger than that of f.sp.sojae, 2.9 and 2.1 × 10¹⁰ Da, respectively. The amounts of repetitive sequence and AT-rich region in the nDNA were also larger in f.sp.adzukicola than f.sp.sojae. These results indicate that the nuclear genome organization of the two formae speciales is differentiated.     

Specificity of Proteinase K from Tritirachium album Limber for Synthetic Peptides

The specificity of proteinase K from Tritirachium album Limber was determined using various synthetic peptide substrates. The esterase activity against N-acylated amino acid esters indicated that the enzyme is primarily specific against aromatic or hydrophobic amino acid residues at the carboxyl side of the splitting point. Secondary interaction for hydrolysis was also studied using peptide esters or others, which showed that the enzyme activity is markedly promoted by elongating the peptide chain to the N-terminal from the splitting point. Thus, peptide chloromethyl ketone derivatives such as Cbz-Ala-Gly-PheCH₂Cl inactivated the enzyme activity markedly.     

Distal Lateral Subungual Onychomycosis Owing to <Emphasis Type="Italic">Tritirachium oryzae</Emphasis>: A Bystander or Invader?

The genus Tritirachium is a mitosporic fungus which inhabits in soil and decaying plant material and also a notable insect pathogen. Human infections with Tritirachium species though rare were previously reported to cause corneal ulcers, otomycosis, onychomycosis, and dermatomycosis of the scalp and hence may be considered as a potential pathogen. Here we report a case of distal lateral subungual onychomycosis involving right great toenail in a 22-year-old female, wherein direct potassium hydroxide preparations, fungal cultures, and molecular sequencing of the isolate established Tritirachium oryzae as the etiological agent. Antifungal susceptibility performed by the microbroth technique of CLSI revealed increased MICs to amphotericin B and low MICs to azoles and echinocandins. The case was managed with surgical nail avulsion followed by topical application of 2% ketoconazole cream resulting regrowth of normal nail. To the best of our knowledge, this is the first report of non-dermatophytic mold T. oryzae causing onychomycosis in India.     

Study of the conidial development and cleistothecium-like structure of some strains of Fonsecaea pedrosoi

The reproduction of twenty-seven strains of Fonsecaea pedrosoi was studied. The denticulate type (asexual reproduction) showed three morphological variations: medium-size, long and sessile forms. These forms can be used to characterize the different strains isolated especially for epidemiological purposes.Only one strain (MR 1335) isolated from a chromomycosis case in Martinique showed a cleistothecium-like structure. This formation was compared to that observed in Phialophora verrucosa, another agent of chromomycosis, and to the cleistothecium of Dictyotrichiella mansonii, the perfect state of Wangiella mansonii which is a saprophyte fungus.     

Biodegradability and biodegradation of poly(lactide)

Poly(lactide) (PLA) has been developed and made commercially available in recent years. One of the major tasks to be taken before the widespread application of PLA is the fundamental understanding of its biodegradation mechanisms. This paper provides a short overview on the biodegradability and biodegradation of PLA. Emphasis is focused mainly on microbial and enzymatic degradation. Most of the PLA-degrading microorganisms phylogenetically belong to the family of Pseudonocardiaceae and related genera such as Amycolatopsis, Lentzea, Kibdelosporangium, Streptoalloteichus, and Saccharothrix. Several proteinous materials such as silk fibroin, elastin, gelatin, and some peptides and amino acids were found to stimulate the production of enzymes from PLA-degrading microorganisms. In addition to proteinase K from Tritirachium album, subtilisin, a microbial serine protease and some mammalian serine proteases such as α-chymotrypsin, trypsin, and elastase could also degrade PLA.     

Predisposition of Aster to Phialophora wilt

The susceptibility of Aster novi-belgii to infection by the vascular wilt pathogen Phialophora asteris varied according to the growing conditions to which the host was subjected before inoculation. Relatively low air temperature, high soil moisture, and soil nutrients rich in potassium or phosphorus were pre-inoculation conditions in which Aster became relatively susceptible. Increased resistance was favoured by growing plants at relatively high air temperatures, low soil moisture, and soil nutrients rich in nitrogen or calcium but low in potassium or phosphorus prior to inoculation.     

Production de deux glycopeptides par le Phialophora cinerescens (Wr.) van beyma cultive in vitro

Résumé Le Phialophora cinerescens alcalinise nettement le milieu liquide sur lequel il est cultivé durant 7 semaines; Le saccharose fourni est rapidement hydrolysé en glucose et fructose. La production des deux glycopeptides neutre et anodique — qui affectent la croissance des jeunes plantes d'Oeillet — a été appréciée. Le glycopeptite neutre est synthétisé dès le début de la culture, le glycopeptide anodique plus tardivement; les variations de la croissance du P. cinerescens cultivé in vitro suggèrent que les deux glycopeptides ne sont pas des produits de l'autolyse du mycélium.

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In vitro growth of the fungus Phialophora cinerescens was studied for 7 weeks. During this period the pH values were shown to increase in the culture filtrates; the added sucrose was rapidly hydrolyzed into glucose and fructose. The production of both neutral and anodic glycopeptides — which are know to affect the growth of carnation seedlings — was periodically monitored. The neutral glycopeptide was synthesized at the beginning of the culture period, the anodic one was synthesized later. The P. cinerescens growth curves indicated these two metabolites were probably not autolytic products of fungal cells.

     

Diversity,Molecular Phylogeny,and Bioactive Potential of Fungal Endophytes Associated with the Himalayan Blue Pine (Pinus wallichiana)

In this study, we investigated the diversity of fungal endophytes associated with Pinus wallichiana from the Western Himalayas, with emphasis on comparison of endophytic communities harbored by the stem and needle tissues of the host and their antimicrobial potential. A total number of 130 isolates, comprising of 38 different genera, were recovered from 210 fragments of the plant. Among the isolated fungi, only a single isolate, Tritirachium oryzae, belonged to the Phylum Basidiomycota whereas the rest belonged to Ascomycota. Dothideomycetes was the dominant class with the highest isolation frequency of 49.2 %. The most frequent colonizers of the host were Alternaria spp., Pestalotiopsis spp., Preussia spp., and Sclerostagonospora spp. The diversity and species richness were higher in needle tissues than in the stems. Antimicrobial activities were displayed by extracts from a total number of 22 endophytes against one or more pathogens. Endophytes designated as P1N13 (Coniothyrium carteri), P2N8 (Thielavia subthermophila), P4S6b (Truncatella betulae), P7N10 (Cochliobolus australiensis), and P8S4 (Tritirachium oryzae) were highly active against Candida albicans. Broad spectrum antimicrobial activities were obtained with the extracts of P8-S4 (Tritirachium oryzae) and P5-N26 (Coniochaeta gigantospora) that were potentially active against the Gram-positive and Gram-negative bacteria as well as the fungal pathogen, Candida albicans. The most prominent antagonistic activity against fungal pathogens was shown by P8-S4 (Tritirachium oryzae), P5-N31a (Truncatella spadicea), and P5-N20 (Fusarium larvarum). Our findings indicate that Pinus wallichiana harbors a rich endophytic fungal community with potential antimicrobial activities. Further studies are needed to understand the ecology and evolutionary context of the associations between the Himalayan pine and its endophytes.     

Removal of ABS from solutions by a common fungus of sewage

Conclusions Phialophora jeanselmei (Margarinomyces heteromorphum) is very common in sewage treatment systems, and can remove ABS from solution even in the absence of glucose. Because glucose is present in sewage treatment systems either as a pollutant or as a byproduct of the metabolism of other organisms, it appears that, to some extent, ABS is removed by this organism. Additional work is needed to determine whether this organism can be made a nucleus of an artificial population deliberately established to act upon some special types of wastes; whether a mononucleate-monospore culture capable of more rapid and more complete removal of ABS can be developed; and what pathways of metabolism and decomposition of ABS might be involved with the idea of breaking into the chains of reaction with other organisms.In the presence of added glucose,Phialophora jeanselmei was found to be capable of removing ABS from shaken and still culture solutions containing an ABS concentration of 10 ppm. More adsorption on the mycelium took place in still culture than in aerated culture. In shaken culture a greater loss of ABS from solution resulted; this may be due to metabolic processes since the ABS was neither found in solution nor adsorbed on the mycelium. While wild cultures are capable of a demonstrable amount of activity, culture efficiency could be improved if selected monospore-mononucleate cultures were developed.