Specific detection of fusarium species in blood and tissues by a PCR technique.

Fusarium species are opportunistic nosocomial pathogens that often cause fatal invasive mycoses. We designed a primer pair that amplifies by PCR a fragment of a gene coding for the rRNA of Fusarium species. The DNAs of the main Fusarium species and Neocosmospora vasinfecta but not the DNAs from 11 medically important fungi were amplified by these primers. The lower limit of detection of the PCR system was 10 fg of Fusarium solani DNA by ethidium bromide staining. To test the ability of this PCR system to detect Fusarium DNA in tissues, we developed a mouse model of disseminated fusariosis. Using the PCR, we detected Fusarium DNA in mouse tissues and in spiked human blood. Furthermore, F. solani, Fusarium moniliforme, and Fusarium oxysporum were testing by random amplified polymorphic DNA (RAPD) analysis. The bands produced by RAPD analysis were purified, cloned, and sequenced. The information was used to design primer pairs that selectively amplified one or several Fusarium species. The method developed may be useful for the rapid detection and identification of Fusarium species both from culture and from clinical samples.

Organisation of the mitochondrial genome of Trichophyton rubrum III. DNA sequence analysis of the NADH dehydrogenase subunits 1, 2, 3, 4, 5 and the cytochrome b gene.

In this paper, we present the nucleotide sequence of a 9761 nt-long segment of the mitochondrial genome of the dermatophyte Trichophyton rubrum that bridges the gap between two previously published segments, making a unique contig that represents approximately 80% of the molecule. The location of all genes on the map is determined except for some tRNA genes expected to flank the LSU rRNA gene not yet sequenced. Starting from the 5' end of the present sequence, we recognized the ND5 and ND2 genes, the cytochrome b gene, an unusually long intergenic spacer of unknown function, as well as the ND3, ND1 and ND4 genes. This sequence extends and confirms the similarity with the mitochondrial genome of Aspergillus nidulans. Interestingly, two cases of partial overlaps between the terminator and initiator codons of successive genes (ND4-ND5 and ND5-ND2) are encountered.

Deep-seated fungal diseases in the South Pacific, especially in New Caledonia.

The main deep-seated fungal diseases and their encountered pathology in New Caledonia and other islands of the South Pacific are reviewed (1970-96). Cryptococcosis is encountered in all islands of the South Pacific, Australia and Papua New Guinea, with a predominance of variety gattii, which is associated with some species of Eucalyptus. Histoplasmosis is not uncommon, and there was an epidemic in New Caledonia in 1994 among people who had visited a bat-inhabited cave. Mycetomas, in particular presenting as pale granules in tissues, are encountered in New Caledonia, Vanuatu, Papua New Guinea, Fiji and French Polynesia. Other fungal infections, such as zygomycosis, sporotrichosis (three cases) and chromomycosis (six cases) are rarely observed in New Caledonia.

First European cases of onychomycosis caused by Onychocola canadensis.

Onychocola canadensis is a new fungus responsible of onychomycosis only isolated until now from Canada and New Zealand. We present the first European cases of onychomycosis caused by this fungus.

Organisation of the mitochondrial genome of Trichophyton rubrum. DNA sequence analysis of the ND4 gene, the ATPase subunit-6 gene, the ribosomal RNA small-subunit gene, the ND6 gene, the COXIII gene, the ATPase subunit-8 gene and six tRNA genes that correspond respectively to the tyrosine, lysine, glutamine, asparagine, isoleucine and tryptophan isoacceptors.

We present the nucleotide sequence of a 5207-bp-long region of the mitochondrial genome of the dermatophyte Trichophyton rubrum. This represents about 1/5th of the total genome and extends a previous study. From the 5' end of the present sequence, the order of genes is as follows: the end of the ND4 gene, the gene coding for subunit 6 of ATPase, the gene coding for the small ribosomal RNA (SSU rRNA), the tyrosyl tRNA gene, the ND6 gene, the COXIII gene, the ATPase 8 subunit gene and a cluster of tRNAs genes corresponding respectively to the lysine, glutamine, asparagine, isoleucine and tryptophan isoacceptors. The interesting features of this region are its compact organisation, the presence of subunit 8 of the ATPase gene and the secondary structure of SSU rRNA which is close to that of Aspergillus nidulans. On the basis of the order of the genes, which is essentially similar to that of A. nidulans, we can also assume that the LSU rRNA subunit gene should be just upstream of this sequenced region.

Subcutaneous abscess caused by Pleurophomopsis lignicola Petr: first case.

The dematiaceous coelomycete Pleurophomopsis lignicola Petr. was isolated repeatedly in pure culture from a subcutaneous abscess of the left leg in a farmer undergoing corticosteroid therapy for asthma gravis. To our knowledge, we report the first case involving this fungus as an aetiological agent of subcutaneous phaeohyphomycosis.

Sex hormone effects on Phialophora verrucosa in vitro and characterization of progesterone receptors.

To determine the effects of sex steroid hormones on the growth of an aetiologic agent of chromoblastomycosis, we studied the dematiaceous fungus Phialophora verrucosa. The in vitro growth of this species on culture media containing either progesterone, testosterone or oestradiol at various concentrations was assessed. Both progesterone and testosterone inhibited the growth of P. verrucosa, whereas oestradiol did not. In other experiments, fungal cytosolic fractions were obtained and steroid binding assays were performed. These studies showed that the presence of progesterone receptors possessed two binding sites as determined by Scatchard analysis, one of which has a high affinity to progesterone (Kd = 6.02 x 10(-8) M) with total binding sites of 120 fmol micrograms-1 protein. These findings suggest that the growth of P. verrucosa is regulated by steroid hormones and that the effect of progesterone could be mediated through fungal intracellular progesterone receptors.

[Chromoblastomycosis in Gabon. Study of 64 cases]. / La chromoblastomycose au Gabon. Etude de 64 cas.

During the last 25 years, 64 cases of chromoblastomycosis were diagnosed in Gabon. They came from the entire territory, where living in rural zones men as well as women were affected. The evolution of the disease has lasted for more than 10 years for the majority of the cases. The preferential location is the lower limb for 78.5% of the patients, with a double localization for one case and an exceptional malignant transformation for another case. Two clinical aspects emerged, despite an apparent polymorphism: extensive patches and warty nodules, spread apart from or adjacent to the others. The histological images were classical and the fumagoid cells, sometimes budding and filamentous, were easily detected during examination of the squama and scabs. Fonsecaea pedrosoï was the only identified species. The 5-fluorocytosine treatment constantly led to an improvement of the lesions. The major problem for these destitute patients is economic, concerning the cost of the therapy.

[Rhino-orbito-cerebral mucormycosis caused by Rhizopus oryzae.A typical case in a cirrhotic patient]. / Mucormycose rhino-orbito-cérébrale due à Rhizopus oryzae. Un cas typique survenu chez une patiente cirrhotique.

The rhino-orbito-cerebral mucormycosis is an acute rapidly progressive fungal infection. This disease is caused by a zygomycetes fungus, most often from the Rhizopus genus. This fungus is saprophitic of the nasal cavity and paranasal sinuses. It becomes pathogenic in some particular conditions, specially during diabetes mellitus. Histopathological study is the only method allowing the diagnosis, by revealing the tissue invasion by characteristic hyphae. Mycologic study allows a definitive identification of the fungus. The authors report a case of rhino-orbito-cerebral mucormycosis in a 44 year-old woman with cirrhosis. She presented an acute blindness and ophthalmoplegia. Despite of a rapid histologic diagnosis from the nasal and ethmoidal biopsies, the patient died 3 days after.

Mitochondrial DNA sequence analysis of the cytochrome oxidase subunit I and II genes, the ATPase9 gene, the NADH dehydrogenase ND4L and ND5 gene complex, and the glutaminyl, methionyl and arginyl tRNA genes from Trichophyton rubrum.

In this paper, we present the nucleotide sequence of a 5248 bp-long region of the mitochondrial (mt) genome of the dermatophyte Trichophyton rubrum. This region which represents about 1/4 of the total mt genome of this species reveals a compact organization of genes including: the glutaminyl tRNA, the methionyl tRNA, the cytochrome oxidase subunit I gene, the arginyl tRNA, the mitochondrial version of the ATPase subunit 9 gene, the cytochrome oxidase subunit II gene and a part of the NADH dehydrogenase ND4L and ND5 gene "complex". The main features of the part of mt DNA sequenced is the non-interrupted COXI gene and the presence in the mitochondrial version of the ATPase 9 gene of a small group IA intron. The extensive amino-acid sequence similarity with the equivalent gene in Aspergillus nidulans and Neuropora crassa indicates that this gene codes for a dicyclohexylcarbodiimide binding protein. The conserved arrangement of this portion of the mt genome and the presence of tRNAs between the protein-coding genes are compatible with a large polycistronic transcript processed by the excision of tRNAs, or similar secondary structures, as proposed for other fungal or mammalian mt DNAS.

Hendersonula toruloidea as an agent of mycotic foot infection in Gabon.

Pathogenic fungi were isolated from 149 of 210 Gabonese patients with clinical signs of tinea pedis. Hendersonula toruloidea was isolated from 51 patients (34.2%), either alone (in 25%) or in association with a dermatophyte species or Candida albicans. Clinical signs of H. toruloidea infection closely resembled those of dermatophyte infection except that hyperkeratinization of the soles was most often associated with infection by H. toruloidea. H. toruloidea mycelium could be readily distinguished from dermatophyte mycelium on direct examination of skin scrapings. All the H. toruloidea isolates closely resembled the Type 1 isolates described by Moore, both microscopically and macroscopically. The importance of establishing a mycological diagnosis of this infection and the therapeutic implications of infection with H. toruloidea are discussed. This fungus was isolated from 34% of our patients from Gabon with clinical tinea pedis.

Maxillary sinusitis from Microascus cinereus and Aspergillus repens.

Microascus was associated with Aspergillus repens in a left maxillary sinus. Tissue contained septale filaments of two types, conidia, ostiolate perithecia containing ascospores corresponding to Microascus cinereus which was identified by culture. The abundance of sexual fructifications in the tissue indicates that pathogenicity is due to Microascus cinereus.

[Fungal otitis in Libreville. Study of 83 cases]. / Les otites mycosiques à Libreville. Etude de 83 cas.

83 cases of mycotic otitis of external ear are reported during a period of 27 months from three departments of otorhinolaryngology in Libreville (Gabon, Central Africa) Prevalence is estimated at about 25% among all infectious otitis. The main functional signs are pruritus, otorrhea, pain and hypoacousia. The physical examination shows masse of white, grey, black or creamy caseous debris, invading the external auditory meatus (EAM) which is sometimes inflammatory. Fungal species responsible of otitis are Aspergillus (54%), yeasts (45%) mainly Candida, infrequently Fusarium (1%). A niger (26%), A. flavus (17%), Candida parapilosis (18%), Candida albicans (9%) are predominant species isolated (70%) among all the 21 species of identified fungi from otomycosis in Gabon. Therapy, done by thorough washing of the ear followed by insertion into the EAM of a wick soaked in Econazole or Amphotericin B, is quickly effective.

Immunochemical characterisation of antigens and growth inhibition of Fonsecaea pedrosoi by species-specific IgG.

Antigens of Fonsecaea pedrosoi, the most common agent of chromomycosis, were characterised by immunoprecipitation with a rabbit antiserum raised against the cell-protein extract and serum from an infected patient. Thirteen antigens were commonly detected and, as some of these antigens could be iodinated, they may be present in the fungal cell wall. Purified IgG from the rabbit antiserum was shown to produce a 50-60% inhibition of fungal growth. Some of the antigens characterised may be important in relation to the stimulation of protective immunity against chromomycosis.