Luciana da Silva Ruiz

Yeasts isolated from blood and catheter in children from a Public Hospital of São Paulo, Brazil

The great majority of nosocomial fungal infections, especially fungemias, are caused by yeasts, mostly of the genus Candida. In addition, such infections may be associated with intravascular catheters. In this study, 80 yeast strains were isolated from hospitalized children, being 59 from blood cultures and 21 from vascular catheter cultures. The prevalent species in both blood and catheter was C. parapsilosis (32.2% and 48.9%, respectively), followed by C. albicans (16.9% and 28.6%, respectively). Concerning enzyme production, 78.8% of the 80 isolates presented strong proteolytic activity but 78.8% showed no phospholipase activity. We also detected two prevalent “killer” biotypes: 511 and 888. Additionally, in five patients, it was possible to observe that the yeast species, “killer” biotype and proteolytic and phospholipase activity of blood and catheter were similar. In view of this, we suggest a transmission of nosocomial yeast infection from catheter to blood.

Nosocomial infection in newborns by Pichia anomala in a Brazilian intensive care unit

Disseminated candidiasis is the most common nosocomial fungal infection, and Candida albicans has been reported to account for 50% to more than 70% of cases of invasive candidiasis. However, recent reports have also suggested the emergence of infections caused by non-albicans species. In addition, less-common pathogenic yeasts (Malassezia, Trichosporon, Rhodotorula, Debaryomyces and Pichia) have recently been reported, with increased frequency, as causes of nosocomial infections with high mortality. This article describes two cases of fungemia caused by Pichia anomala in newborns that occurred in an intensive care unit (ICU), in November 2004 at the Instituto da Criança (Pediatric Institute) of the Hospital das Clínicas of the School of Medicine, São Paulo University, Brazil. The principal factors related to virulence (proteinase and phospholipase) and the susceptibility of the isolated strains to antifungal agents were also evaluated, and the biotype of each strain was determined through the use of an epidemiological marker (killer biotype).

Candiduria in a public hospital of São Paulo (1999-2004): characteristics of the yeast isolates

The study involved 100 yeast isolates, obtained from urine samples provided by a Public Pediatric Hospital of São Paulo, Brazil, from 1999 to 2004. The most frequent species was Candida albicans, followed by C. tropicalis, C. glabrata and C. parapsilosis. In regard to virulence, 97% of the isolates showed index 3 for proteinase and 63% index 2 for phospholipase. The most frequent killer biotypes were 511 and 888.

Molecular identification of Candida dubliniensis isolated from oral lesions of HIV-positive and HIV-negative patients in São Paulo, Brazil

Candida dubliniensis is a new, recently described species of yeast. This emerging oral pathogen shares many phenotypic and biochemical characteristics with C. albicans, making it hard to differentiate between them, although they are genotypically distinct. In this study, PCR (Polymerase Chain Reaction) was used to investigate the presence of C. dubliniensis in samples in a culture collection, which had been isolated from HIV-positive and HIV-negative patients with oral erythematous candidiasis. From a total of 37 samples previously identified as C. albicans by the classical method, two samples of C. dubliniensis (5.4%) were found through the use of PCR. This study underscores the presence of C. dubliniensis, whose geographical and epidemiological distribution should be more fully investigated.

Genotyping by RAPD-PCR analyses of Malassezia furfur strains from pityriasis versicolor and seborrhoeic dermatitis patients.

Malassezia furfur is lypophilic yeast commonly associate with dermatological disorders. In the present work, we described the isolation of 47 M. furfur strains from three groups of patients: pityriasis versicolor (21 isolates), seborrhoeic dermatitis (15 isolates) and seborrhoeic dermatitis of the HIV positive patients (11 isolates). To investigate the identity of the strains at molecular level, DNA genomic of M. furfur strains were prepared and used to RAPD-PCR analyses. RAPD assay were carried out using two decamer primers and bands pattern generated were analyzed by an Unweighted Pair-Group Method (UPGMA). Dendrogram established a distinct differentiation between M. furfur isolates from pityriasis versicolor and seborrhoeic dermatitis patients with or without AIDS. We concluded that RAPD typing presented a high discriminatory power between strains studied in this work and can be applied in epidemiological investigation of skin disease causing by M. furfur.

Candida isolated from vaginal mucosa of mothers and oral mucosa of neonates : Occurrence and biotypes concordance

Background: The common occurrence of Candida spp. on the vaginal mucosa of pregnant women suggests this as the source of neonatal candidiasis. Methods: This study investigated the occurrence of yeasts on the vaginal mucosa of 100 mothers at the time of birth, and on the oral mucosa of their respective neonates, all full-term, on the 1st, 3rd, and 9th days after birth by vaginal (72 cases) and cesarean (28 cases) routes. In each case where concordance at the level of species was found between the isolate from the mother and that from the neonate, tests were made to check for concordance between the genotypic and phenotypic profiles (susceptibility to killer toxins, serotyping, proteinase and phospholipase production, and susceptibility to antifungal agents). Results: For the vaginal-route group, yeasts were recovered from the vaginal mucosa of 47.2% of the mothers and from 25% of the neonates. For the cesarean-route group, these rates were 46.4% and 3.6%, respectively. Species found most frequently in the samples from the mothers and the neonates were, respectively C. albicans and C. guilliermondii. For the vaginal-route group, the rate of mother/neonate concordance at the level of species was 23.5% and no cases of concordance for the cesarean births. Of these cases with species concordance, there was concordance between the genotypic and phenotypic profiles in 6% (2 cases). Conclusion: The vaginal mucosa was not the main route of transmission of the Candida species to the neonate, because there was concordance between the genotypic and phenotypic profiles in only 6% (2 cases).

FUNGEMIA CAUSED BY Candida SPECIES IN A CHILDREN'S PUBLIC HOSPITAL IN THE CITY OF SÃO PAULO, BRAZIL: STUDY IN THE PERIOD 2007-2010

Candidemia remains a major cause of morbidity and mortality in the health care environment. The epidemiology of Candida infection is changing, mainly in relation to the number of episodes caused by species C. non-albicans. The overall objective of this study was to evaluate the frequency of yeasts of the genus Candida, in a four-year period, isolated from blood of pediatric patients hospitalized in a public hospital of the city of São Paulo, Brazil. In this period, yeasts from blood of 104 patients were isolated and, the identified species of Candida by phenotypic and genotypic methods were: C. albicans (39/104), C. tropicalis (25/104), C. parapsilosis (23/104), Pichia anomala (6/104), C. guilliermondii (5/104), C. krusei (3/104), C. glabrata (2/104) and C. pararugosa (1/104). During the period of the study, a higher frequency of isolates of C. non-albicans (63.55%) (p = 0.0286) was verified. In this study we verified the increase of the non-albicans species throughout the years (mainly in 2009 and 2010). Thus, considering the peculiarities presented by Candida species, a correct identification of species is recommended to lead to a faster diagnosis and an efficient treatment.

Dermatophytosis and immunovirological status of HIV-infected and AIDS patients from Sao Paulo city, Brazil

Over the past decades, more people became infected with human immunodeficiency virus (HIV) and developed acquired immunodeficiency syndrome (AIDS). Because of that the incidence of fungal infections rose dramatically. It happened because this virus can modify the course of fungal diseases, leading to altered clinical pictures. The aim of this study was to evaluate epidemiological and biological aspects of dermatophytosis in HIV‐positive and AIDS patients living in the city of São Paulo, Brazil. A total of 84 (44 HIV‐positive and 40 AIDS) patients were enrolled in this study. The patients were tested for dermatophyte infections, as well as for the CD4+/CD8+ and HIV viral load counts. Tinea unguium was most frequently observed in AIDS patients, whereas Tinea pedis was mostly observed in HIV‐positive patients. The most frequent dermatophyte species was Trichophyton rubrum. CD4+ counts and CD4+/CD8+ ratios were not associated with a higher risk for dermatophytosis. On the other hand, viral load higher than 100 000 copies/ml was associated with a higher frequency of dermatophytosis. The results suggest to that although dermatophytosis is common in HIV‐positive and AIDS patients, the degree of immunosuppression does not seems to correlate with increased risk of this fungal infection. In addition, high viral load as a predictive risk factor for dermatophyte infection should be subject of further evaluations.

Oral colonization: a possible source for candidemia in low-weight neonates.

OBJECTIVE To check the oral colonization in neonates at high-risk and to associate these cases with candidemia. SUBJECTS AND METHODS This study was conducted in the NICU. For six months, 125 high-risk neonates were investigated for oral colonization and septicemia by yeasts. From this total, 19 neonates had yeasts on the oral mucosae and 12 neonates developed fungemia. All of the 12 neonates with fungemia were included in the amount of 19 who have presented oral colonization by yeasts. RESULTS There was a species concordance between the yeasts of the oral mucosae and the blood in 6 neonates (50%) among the 12 neonates with oral colonization and septicemia at the same time. The yeasts isolated in these 6 cases regarding the species concordance were Candida albicans (5 cases, 83.4%) and Candida parapsilosis (1 case, 16.6%). All of the cases involving an association were confirmed by PFGE. All of the strains of yeasts involved in oral colonization and also blood presented the same karyotype. A total of 66.6% of the patients with strains in agreement progressed to death. CONCLUSION The results indicate the relevance of monitoring the oral microbiota, as a possible source of fungal infection, and assisting to develop appropriate therapeutic strategy.

Trichophyton rubrum Isolated from AIDS and Human Immunodeficiency Virus-Infected Patients in São Paulo, Brazil: Antifungal Susceptibility and Extracellular Enzyme Production

Background: In order to identify intraspecific variations in Trichophyton rubrum and to correlate them to the immunological status of the host, sixty strains isolated from AIDS, HIV-positive and HIV-negative patients were compared for the production of extracellular enzymes and for their susceptibility to several antifungal drugs. Methods: The isolates were tested for their ability to secrete keratinases, proteinases, phospholipases, lipases and DNases. Likewise, we investigated their susceptibility to amphotericin B, ketoconazole, ciclopiroxolamine, griseofulvin, miconazole and tolnaftate. Results: Variations in the Minimal Inhibitory Concentration (MIC80) values were observed for all antifungals tested, but they were similarly distributed among the three clinical groups. Griseofulvin showed the most prominent differences among the three groups of isolates. Regarding enzyme secretion, all samples secreted keratinases and DNases, while none secreted phospholipases. Proteinases and lipases were secreted by some of them. Conclusions: The differences among isolates of the three groups were not statistically significant and therefore could not be ascribed to a given clinical status. Intraspecific variations similarly occurred in each group, irrespective of the immunological status of the patients.