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.

[Egg crater mattresses: a deposit of methicillin-resistant staphylococcus aureus?]. / Colchões do tipo caixa de ovo: um reservatório de Staphylococcus aureus resistente à meticilina?

The purpose of this study was to evaluate the microbiological conditions of the egg crater mattress in hospital use to identify the presence of Staphylococcus aureus and its methicillin-resistance phenotype (MRSA). Petrifilm plates were used to collect the microbiological data from the mattresses, in pre-established positions. A total 180 plates were collected in 15 mattresses, 139 (72.2%) of which were positive for Staphylococcus aureus. Of the positive plates, 77 (55.4%) were collected before and 62 (44.6%) after washing the mattresses. There was a significant reduction (p=0.023) in Colony Forming Units (CFU); however, regarding the resistance profile, 8 (53.3%) mattresses with MRSA were identified. Results show the risk of these mattresses acting as a secondary deposit in the infection chain, especially regarding the presence of MRSA.

Colchões do tipo caixa de ovo: um reservatório de Staphylococcus aureus resistente à meticilina? / Egg crater mattresses: a deposit of methicillin-resistant staphylococcus aureus? / Colchones del tipo caja de huevos: ¿un reservorio de staphylococcus aureus resistentes a la meticilina?

O estudo teve como objetivo avaliar as condições microbiológicas de colchões caixa de ovo em uso hospitalar com a finalidade de identificar a presença de Staphylococcus aureus e seu fenótipo de resistência à meticilina (MRSA). Coletaram-se as amostras microbiológicas nos colchões por meio de placas de contato PetrifilmTM em posições pré-estabelecidas. Totalizou-se 180 placas coletadas em 15 colchões, das quais 139 (72,2 por cento) foram positivas para Staphylococcus aureus. Desse total, 77 (55,4 por cento) e 62 (44,6 por cento) corresponderam respectivamente à coleta antes e após a lavagem dos colchões. Evidenciou-se redução significante (p=0,023) das Unidades Formadoras de Colônias (UFC), entretanto com relação ao perfil de resistência foi identificado 8 (53,3 por cento) colchões com MRSA. Diante dos resultados, pode-se inferir sobre o risco destes colchões atuarem como reservatórios secundários na cadeia de infecção, especialmente no que se refere à presença de MRSA.

The purpose of this study was to evaluate the microbiological conditions of the egg crater mattress in hospital use to identify the presence of Staphylococcus aureus and its methicillin-resistance phenotype (MRSA). PetrifilmTM plates were used to collect the microbiological data from the mattresses, in pre-established positions. A total 180 plates were collected in 15 mattresses, 139 (72.2 percent) of which were positive for Staphylococcus aureus. Of the positive plates, 77 (55.4 percent) were collected before and 62 (44.6 percent) after washing the mattresses. There was a significant reduction (p=0,023) in Colony Forming Units (CFU); however, regarding the resistance profile, 8 (53.3 percent) mattresses with MRSA were identified. Results show the risk of these mattresses acting as a secondary deposit in the infection chain, especially regarding the presence of MRSA.

El estudio tuvo como objetivo evaluar las condiciones del tipo caja de huevos en uso hospitalario con la finalidad de identificar la presencia de Staphylococcus aureus y su fenotipo de resistencia a la meticilina (MRSA). Se recolectaron las muestras microbiológicas en los colchones a través de placas de contacto PetrifilmTM en posiciones preestablecidas. Se totalizaron 180 placas colectadas en 15 colchones, de las cuales 139 (72,2 por ciento) fueron positivas para Staphylococcus aureus. De ese total, 77 (55,4 por ciento) y 62 (44,6) correspondieron respectivamente a la recolección anterior y posterior al lavado de los colchones. Se evidenció reducción significativa (p=0,023) de las Unidades Formadoras de Colonias (UFC), mientras que con relación al perfil de resistencia se identificaron 8 (53,3 por ciento) colchones con MRSA. Ante tales resultados, se puede inferir el riesgo de que estos colchones actúen como reservorios secundarios en la cadena de infección, especialmente en lo que se refiere a la presencia de MRSA.

[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.

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

OBJETIVO: caracterizar fenotipicamente leveduras isoladas do conteúdo vaginal de 223 mulheres adultas, sintomáticas (S) e assintomáticas (A) para vulvovaginite, e determinar os indicadores clínicos que possivelmente levam ao surgimento de sinais e sintomas relacionados ao acometimento da mucosa por essa patologia. MÉTODOS: inicialmente foi aplicado um questionário, com questões abertas e fechadas, sobre dados clínicos epidemiológicos. Logo, ocorreu o diagnóstico micológico com semeadura em meio Chrom Agar Candida, identificação micromorfológica e bioquímica. Métodos específicos para detecção de fatores de virulência, proteinase e fosfolipase foram empregados. A análise estatística das variáveis foi estabelecida utilizando os testes χ2 e χ2 de Pearson. RESULTADOS: Candida albicans foi a espécie mais prevalente (87%, S e 67%, A), seguida de Candida glabrata (4%, S e 17%, A). O número de mulheres que referiram adoção de anticoncepcionais foi mais alto entre as sintomáticas, 77%. Nos dois grupos estudados, em torno de 87% apresentaram ciclos menstruais regulares, 57% das mulheres eram casadas com idade entre 30 a 40 anos. Em relação a práticas sexuais, houve para parte das pacientes, concomitância entre os hábitos, anal, oral e vaginal. Em relação à fosfolipase, apenas Candida albicans produziu este fator de virulência em 37,5%. A proteinase foi detectada em Candida albicans, Candida glabrata e Candida parapsilosis. Esse último fator de virulência esteve associado, principalmente, a isolados de pacientes sintomáticas. CONCLUSÕES: a colonização e infecção da mucosa vaginal por levedura é real com diversas espécies de Candida presentes. No entanto, Candida albicans se destaca como espécie prevalente em mucosa vaginal de mulheres adultas. Fica evidente a emergência de espécies de Candida não albicans, algumas com resistência intrínseca aos azólicos, tais como Candida glabrata, Candida...

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 χ2 and Pearson’s χ2 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.

[Onychomycosis: clinical, epidemiological and mycological study in the municipality of São José do Rio Preto]. / Onicomicose: estudo clínico, epidemiológico e micológico no município de São José do Rio Preto.

This was a clinical-epidemiological and mycological study on 184 patients at the university hospital: 200 samples, 142 positive samples, 98 yeasts and 68 filamentous fungi. Candida parapsilosis (47%) and Trichophyton rubrum (38%) were prevalent. They were 100% sensitive to ketoconazole and 99% sensitive to amphotericin B. The highest prevalences were among women (80%), adults (62%) and toes (84%).

Onicomicose: estudo clínico, epidemiológico e micológico no município de São José do Rio Preto / Onychomycosis: clinical, epidemiological and mycological study in the municipality of São José do Rio Preto

Estudo clínico-epidemiológico e micológico em 184 pacientes de Hospital Escola: 200 amostras, 142 positivas, 98 leveduras e 68 fungos filamentosos. Candida parapsilosis (47 por cento) e Trichophyton rubrum (38 por cento) foram prevalentes. Ao cetoconazol, 100 por cento de sensibilidade e a anfotericina B, 99 por cento. Prevalência em mulheres (80 por cento), adultos (62 por cento) em pododáctilos (84 por cento).

This was a clinical-epidemiological and mycological study on 184 patients at the university hospital: 200 samples, 142 positive samples, 98 yeasts and 68 filamentous fungi. Candida parapsilosis (47 percent) and Trichophyton rubrum (38 percent) were prevalent. They were 100 percent sensitive to ketoconazole and 99 percent sensitive to amphotericin B. The highest prevalences were among women (80 percent), adults (62 percent) and toes (84 percent).