Comparative genomics of opportunistic Phialophora species involved in divergent disease types.

BACKGROUND: Black opportunists Phialophora verrucosa complex species can cause different disease types in competent and in immunocompromised individuals, but are remarkably overrepresented in CARD9-related infections. OBJECTIVES: To better understand the ecology and potential pathogenicity of opportunistic Phialophora species and reveal eventual genetic parameters associated with the behaviour in vivo and genetic profiles in patients with CARD9 immunodeficiency. METHODS: Genomes of 26 strains belonging to six species of the Phialophora verrucosa complex were sequenced. Using multilocus analysis, all environmental and clinical strains were identified correctly. We compared the genomes of agents from different disease types among each other including CARD9 immunodeficiency. RESULTS: We obtained genome sizes of the 26 Phialophora strains ranged between 32 and 37 MB. Some species showed considerable intraspecific genomic variation. P americana showed the highest degree of variability. P verrucosa was variable in CAZy enzymes, whereas P americana varied in PKS-related genes. Phialophora species, particularly P verrucosa, are relatively frequent in patients with CARD9-related immunodeficiency. Different mutations in the CARD9 gene seem to increase susceptibility for infection by different groups of species, that is either Candida, dermatophytes or black fungi. A number of patients with chromoblastomycosis revealed an as yet unknown CARD9 mutation. TNFα impairment was prevalent in patients with CARD9 infections, while CBM patients were invariably IFNγ. CONCLUSIONS: From genomic investigations, the known virulence factors between clinical and environmental strains did not reveal any significant difference. Phialophora complex has an equal chance to cause infection in humans, either healthy or CARD9-impaired.

Phaeohyphomycosis: Cytomorphologic Evaluation in Eleven Cases.

OBJECTIVE: Phaeohyphomycosis caused by phaeoid fungi is a type of mycosis emerging worldwide which causes a wide variety of clinical manifestations. STUDY DESIGN: A retrospective analysis of 11 cases diagnosed with fungal inflammation on cytology over a period of 6 years (2013-2018) was done along with culture/histopathologic confirmation. RESULTS: Of the total of 11 cases, 9 cases presented with subcutaneous swellings and 1 case each with brain and lung lesions. The age range was 30-83 years (mean: 53.6); 8 patients were male and 3 were female. Cytologic smears showed fungal profiles with septate tortuous hyphae, as well as swollen and narrow, yeast-like swellings with an irregular breadth of the hyphae in all cases. The fungal profiles were visualized on a Masson-Fontana stain. The background showed inflammatory cells, giant cells, and necrosis in variable proportions. Five cases were diagnosed as phaeohyphomycosis on cytology, whereas 3 cases were misdiagnosed as aspergillus and 2 as candida. In 1 case, typing of the fungus was not done. Histopathology was available in 5 cases, and in all these a diagnosis of phaeohyphomycosis was reached. Ten of the 11 cases had confirmation on fungal culture. CONCLUSIONS: Phaeoid fungi are rarely seen in routine cytologic practice. Careful evaluation of cytologic smears and an awareness of the characteristic morphologic features of phaeohyphomycosis are helpful in arriving at a correct diagnosis. Fine needle aspiration cytology provides a rapid diagnosis, enabling prompt therapy.

Visceral Mycoses in Autopsied Cases in Japan from 1989 to 2013.

BACKGROUND AND METHODS: Our group has continuously studied the epidemiology of visceral mycoses (VM) among autopsy cases in Japan from 1989 to 2013. RESULTS: First, from a total of 11,149 autopsied cases, 571 (5.1%) cases of VM were observed in 2013. It was significantly higher than those of 2005 (p < 0.05) and earlier. Notably, incidence of cases with mucormycetes (Muc) in 2013 was higher than that of 1997 and earlier (p < 0.001), especially in leukemia cases. Muc cases also showed higher rate of "severe infection" compared with other cases (p < .0001). Emerging diseases were also observed. Severe fever with thrombocytopenia syndrome cases showed high incidence of VM as a complication. In addition, we observed cases with the rare mycoses caused by Phialopohra verrucosa and Rhodotorula spp. in our analysis. Moreover, the predominant fungal agent of central nervous system infections changed from Cryptococcus spp. to Aspergillus spp. in 2013. This may be considered a breakthrough infection. CONCLUSION: The prevalence of VM in 2013 became higher than those of 2005 (p < 0.05) and earlier, with a notable increase of incidence in cases with Muc. The occurrence of breakthrough VM and emerging mycoses deserve attention.

Chromoblastomycosis--a clinical and mycological study of 71 cases from Sri Lanka.

Chromoblastomycosis, a well-documented chronic fungal infection, represents a specific clinical entity with typical warty cutaneous nodules and a worldwide distribution. Although more prevalent in tropical and subtropical regions, only a few reports are available from Sri Lanka or from Asia. Five etiologic agents of chromoblastomycosis have been recognized worldwide. Of these the majority of infections have been caused by Fonsecaea pedrosoi. During the period from 1952 to 1962, only twelve culturally proven cases of this disease had been recorded from Sri Lanka. The fungus responsible was F. pedrosoi. The present report presents a study of the clinical and mycological features of 71 Sri Lankan patients with chromoblastomycosis for the 16-year period from 1978 to 1993. It documents three etiological agents. Culture identification was made in 69 cases. The three fungal species were Fonsecaea pedrosoi (64), Phialophora verrucosa (3) and a fungus compatible morphologically with F. compacta (2). The isolation of a fungus morphologically compatible with F. compacta is of significance since only 12 cases have been documented in the world's literature so far.

Identification of dematiaceous fungi and their role in human disease.

Phaeohyphomycosis is the disease caused by the dematiaceous hyphomycetes or those fungi that are darkly pigmented because of the presence of melanin in their cell wall. Infections with these fungi may remain localized at the site of traumatic inoculation or within the sinuses or may become disseminated. As these fungi pose a significant problem to both the clinician who must treat the patient and the laboratorian faced with identification of the organism, a selection of representative organisms was presented at the meeting entitled "Focus on Fungus Infections." These fungi often infect patients who are not overtly immunocompromised and therefore may be encountered in a wide variety of cases.

Cromoblastomicose auricular: relato de um caso / Auricular chromoblastomycosis : case report

E apresentado um caso de cromoblastomicose auricular apresentado sob forma de lesao eritemato-escamosa, tendo tido o diagnostico de eczema. Os AA discutem sobre a raridade desta localizacao e aspecto clinico. Embora os poucos casos relatados de cromoblastomicose auricular tenham sido causados pela Fonsecaea pedrosoi, neste o agente etiologico foi a Phialophora verrucosa.

Isolation of Lecythophora mutabilis and Wangiella dermatitidis from the fruit eating bat, Eidolon helvum.

Lecythophora mutabilis was isolated from the lungs of 3 and from the liver of 2 bats, Eidolon helvum a fruit eating species. Wangiella dermatitidis was recovered from the liver of 2 bats of the same species. The isolates were pathogenic for laboratory mice when injected by subcutaneous, intraperitoneal and intravenous routes. W. dermatitidis was neurotropic in the mice injected intravenously.

Lipases of Fonsecaea pedrosoi and Phialophora verrucosa.

Lipase activity was demonstrable titrimetrically in the culture filtrates of Fonsecaea pedrosoi and Phialophora verrucosa on the 6th day of incubation reaching a peak on the 15th and 12th days respectively for the two fungi. Purified lipases of F. pedrosoi and P. verrucosa, with specific activities of 36.0 and 39.4 fold increase respectively, were obtained by cold acetone extraction, gel filtration followed by ion exchange chromatography. The lipases had the same optimum pH (5.5) and temperature (35 degrees C). The molecular weights of the lipases of F. pedrosoi and P. verrucosa, estimated by gel filtration on Sephadex G-100, were 25,000 and 20,000, respectively and the enzymes showed broad substrate specificity. The possible role of lipase in the pathogenesis of infection caused by the fungi is discussed.

The mucilage secreted by roots and its possible role in cell-cell recognition for the adhesion of fungal pathogens to root surfaces of Zea mays L.

The outer cells of the roots of plants secrete a mucilage which lubricates the root and keeps it moist. The mucilage is secreted from the Golgi apparatus in vesicles which fuse at the plasma membrane. In maize roots a complex of at least three polysaccharides and glycoproteins are formed, some of which have a large proportion of fucose in their composition. The synthesis of these compounds can be readily monitored because fucose can be easily identified, and especially because exogenous fucose is not catabolized but is incorporated intact into the polymers. The synthesis of the polymers seems to be initiated in the endoplasmic reticulum in conjunction with polyprenoid oligosaccharides that contain fucose. Lipid-oligosaccharides of nine sugar residues can be obtained from the membrane preparations of the root cells. These compounds are polyprenyl diphosphate derivatives. A GDP-fucose:polyprenyl phosphate transfucosylase occurs in the endoplasmic reticulum, whereas fucosyl transferase that transfers fucose to a polymer occurs mainly in the Golgi apparatus. The indirect evidence suggests that oligosaccharides of polyprenyl diphosphate compounds are transferred to proteins, elaborated in the Golgi apparatus, and large molecular weight polysaccharides are finally exported as the mucus. Part of the mucus is acidic and in some respects resembles pectin. The presence of fucose in such large quantities in maize root mucilage suggested that this might have some significance for the recognition of these plants by parasitic root fungi. The adsorption of mucilage by pathogenic fungi was investigated with two types of fungi, a highly specialized ectotrophic root-infecting fungus, e.g. Phialophora radicicola and a vascular wilt fungus capable of attacking a great variety of tissues, e.g. Fusarium moniliforme. The adsorption of radioactively labelled and fluorescently labelled polymers by the pathogenic fungi was investigated. The character and proportion of fungal surfaces present in vitro were standardised by the production and semi-synchronous germination of populations of conidia. Changes in appearance of fungal walls, present before and after germination, were examined ultrastructurally. There was polyanionic material on hyphal but less on conidial surfaces of the ectotrophic root-infecting fungi. In contrast this material was present to similar extents on both hyphal and conidial surfaces of F. moniliforme.(ABSTRACT TRUNCATED AT 400 WORDS)

Evaluation of proteolytic activity to differentiate some dematiaceous fungi.

A total of 123 isolates of Cladosporium spp., Exophiala spp., Fonsecaea spp., Lecythophora hoffmannii, Phaeoannellomyces werneckii, Phialophora spp., Wangiella dermatitidis, and Xylohypha bantiana were tested for proteolytic activity by using 26 different formulations of gelatin, milk, casein, and Loeffler media. Other physiological properties examined included hydrolysis of tyrosine and xanthine, sodium nitrate utilization in Czapek Dox agar, and thermotolerance. Isolates of Exophiala jeanselmei, Fonsecaea compacta, Fonsecaea pedrosoi, W. dermatitidis, and X. bantiana lacked proteolytic activity. Proteolytic activity was variable among the remaining species, depending on the type of medium used. Thermotolerance had value in distinguishing some taxa.

Studies on pathogenic dematiaceous fungi. 1. Isolation from natural sources.

Two hundred and twenty-six samples of woody materials, vegetable matter and soil were processed by the direct plating and mouse inoculation technique for the isolation of pathogenic dematiaceous fungi. The species of fungi isolated were Fonsecaea pedrosoi - 13, Cladosporium carrionii - 7 and Phialophora verrucosa -4 isolates. The mouse inoculation technique was found to be much better than direct plating for the recovery of these fungi. Woody plant materials proved to be a good sample source for pathogenic dematiaceous fungi contributing about 90% of the isolates. All the isolates were pathogenic for mice as evidenced by the presence of dark nodular lesions containing fungal elements in the organs of experimentally infected animals.

Role of the fungal cell wall in the granulomatous response of mice to the agents of chromomycosis.

Mice were given, intraperitoneally, inocula of a cell-wall preparation and fractions thereof from Fonsecaea pedrosoi, F. compactum, Cladosporium carrioni and Phialophora verrucosum. Large doses of cell-wall preparation, with or without trypsin treatment, produced a pronounced loss of body weight, a granulomatous reaction and, sometimes, death. After extraction of the cell wall preparation with 1N NaOH, three fractions were obtained: an alkali-insoluble fraction 1; an alkali-soluble acid-insoluble fraction 2; and an alkali- and acid-soluble fraction 3. Intravenous administration showed that only fraction 1 induced a granulomatous reaction and death in mice.

Pathogenicity 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 keratitis following penetrating keratoplasty.

A keratitis with an unusual, sessile, filamentary mass extending into the anterior chamber developed in a patient three weeks after penetrating keratoplasty. The causative organism was identified as Exophilia (Wangiella) dermatitidis, a dematiaceous fungus. The infection was cured with a combination of medical and surgical therapy. Inoculation of the isolate into rabbit corneas produced a similar keratitis from which the same organism was cultured. Miconazole levels measured in corneal tissue removed at surgery were approximately 25 times greater than the minimum inhibitory concentration for the fungal isolate.

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.

Subcutaneous chromomycotic abscesses caused by Phialophora gougerotii.

Subcutaneous chromomycotic absesses due to phialophora gougerotii in a 50-year-old woman were characterized by subcutaneous abscesses adhering together through fistulous tracts, rupturing and leaving black crusts over multiple sinuses mimicking mycetoma. Reinfection occurred in contaminated excised lesions. Intradermal inoculation in different anemic patients caused abscesses in 14 days and healed without therapy in eight weeks. No spreading to other organs was found after intraperitoneal inoculation of mice for eight weeks.