International Society of Human and Animal Mycology (ISHAM)-ITS reference DNA barcoding database--the quality controlled standard tool for routine identification of human and animal pathogenic fungi.

Human and animal fungal pathogens are a growing threat worldwide leading to emerging infections and creating new risks for established ones. There is a growing need for a rapid and accurate identification of pathogens to enable early diagnosis and targeted antifungal therapy. Morphological and biochemical identification methods are time-consuming and require trained experts. Alternatively, molecular methods, such as DNA barcoding, a powerful and easy tool for rapid monophasic identification, offer a practical approach for species identification and less demanding in terms of taxonomical expertise. However, its wide-spread use is still limited by a lack of quality-controlled reference databases and the evolving recognition and definition of new fungal species/complexes. An international consortium of medical mycology laboratories was formed aiming to establish a quality controlled ITS database under the umbrella of the ISHAM working group on "DNA barcoding of human and animal pathogenic fungi." A new database, containing 2800 ITS sequences representing 421 fungal species, providing the medical community with a freely accessible tool at http://www.isham.org/ and http://its.mycologylab.org/ to rapidly and reliably identify most agents of mycoses, was established. The generated sequences included in the new database were used to evaluate the variation and overall utility of the ITS region for the identification of pathogenic fungi at intra-and interspecies level. The average intraspecies variation ranged from 0 to 2.25%. This highlighted selected pathogenic fungal species, such as the dermatophytes and emerging yeast, for which additional molecular methods/genetic markers are required for their reliable identification from clinical and veterinary specimens.

Pithomyces species (Montagnulaceae) from clinical specimens: identification and antifungal susceptibility profiles.

The fungal genus Pithomyces comprises numerous dematiaceous saprobic species commonly found on dead leaves and stems of a great variety of plants. Occasionally, they have been recovered from clinical specimens. We morphologically and molecularly (rDNA sequences) investigated a set of 42 isolates tentatively identified as Pithomyces recovered from clinical specimens in the United States. The predominant species were P. chartarum and P. sacchari (33.3% each), followed by Pithomyces sp. I (28.6%) and P. maydicus (4.8%). Most of the isolates were obtained from samples of superficial tissue (50%), the respiratory tract (21.4%), and the nasal region (19%). In general, these fungi were highly susceptible in vitro to the eight antifungal agents tested.

Molecular identification and in vitro response to antifungal drugs of clinical isolates of Exserohilum.

Exserohilum is an agent of human and animal mycoses. Although classification has been based on a few subtle morphological differences, three species of clinical interest have been traditionally accepted. In this study, by using a multigene sequence analysis, we have demonstrated that Exserohilum longirostratum and E. mcginnisii are probable synonyms of E. rostratum. The isolates tested were mainly from the nasal region. Antifungal susceptibility testing demonstrated high activity of the eight agents tested against this fungus.

[Phenotypic characterization of yeasts isolated from the vaginal mucosa of adult women]. / Caracterização fenotípica de leveduras isoladas da mucosa vaginal em mulheres adultas.

PURPOSE: to characterize, phenotypically, yeasts isolated from the vaginal content of 223 symptomatic (S) and asymptomatic (A) adult women with vulvovaginitis, and to determine the clinical indicators which may lead to the appearance of signs and symptoms related to the mucosa involvement by this pathology. METHODS: a questionnaire with open and closed questions on epidemiological clinical data was applied initially. Then, mycological diagnosis with sowing in Chrom Agar Candida was done, followed by micro-morphological and biochemical identification. Specific methods for the detection of the virulence factors, proteinase and phospholipase were employed. Statistical analysis was performed through chi2 and Pearson's chi2 tests. RESULTS: the most prevalent species found was Candida albicans (87%, S and 67%, A) followed by Candida glabrata (4%, S e 17% A). The number of women reporting the use of contraceptives was higher among the symptomatic, 77%. In the two groups studied, about 87% of the women presented regular menstrual cycles and 57% were married with ages between 30 to 40 years old. Concerning the sexual practices, there has been concomitance among anal, oral and vaginal habits from the patients. Only Candida albicans produced the virulence factor phospholipase in 37.5% of them. Proteinase has been detected in Candida albicans, Candida glabrata and Candida parapsilosis. This latter virulence factor was mainly associated to isolates from symptomatic patients. CONCLUSIONS: it is a fact that the vaginal mucosa can be colonized and infected by yeasts, with several Candida species present. Nevertheless, Candida albicans is the most prevalent in the vaginal mucosa of adult women. It is evident the emergence of non-albicans Candida species, some of them with intrinsic resistance to azolics, such as Candida glabrata, Candida parapsilosis, Candida tropicalis, and Candida guillermondii, which can be explained by the inadequate use of medicines and empirical treatment.