Eveline Pipolo Milan

Multicenter Brazilian Study of Oral Candida Species Isolated from Aids Patients

Oropharyngeal candidiasis continues to be considered the most common opportunistic disease in Aids patients. This study was designed to investigate species distribution, serotype and antifungal susceptibility profile among Candida spp. isolated from the oral cavity of Aids patients recruited from six Brazilian university centers. Oral swabs from 130 Aids patients were plated onto CHROMagar Candida medium and 142 isolates were recovered. Yeast isolates were identified by classical methods and serotyped using the Candida Check or target system-Iatron. Antifungal susceptibility testing was performed according to the NCCLS microbroth assay. C. albicans was the most frequently isolated species (91%), and 70% of the isolates belonged to serotype A. We detected 12 episodes of co-infection (9%), including co-infection with both serotypes of C. albicans. Non-albicans species were isolated from 12 episodes, 50% of them exhibited DDS or resistance to azoles. Otherwise, only 8 out 130 isolates of C. albicans exhibited DDS or resistance to azoles. Brazilian Aids patients are infected mainly by C. albicans serotype A, most of them susceptible to all antifungal drugs.

Fusarium Nail and Skin Infection: A Report of Eight Cases from Natal, Brazil

Fusarium spp. are non-dermatophytic hyaline moulds distributed worldwide and recovered from the nature as soil saprophytes and plant pathogens. Human infections are usually precipitated by local or systemic predisposing factors and disseminated infection is associated with impaired immune responses. We report eight cases of cutaneous lesions caused by Fusarium spp. All patients were immunocompetent. Seven cases with presented onychomycosis and one patient with interdigital intertrigo. It is important to alert the medical community about the relevance of the opportunistic fungi, such as Fusarium spp., which have emerged as human infectious agents, emphasizing the importance of correct etiological identification, allowing for appropriate treatment.

Genotyping of methicillin-resistant Staphylococcus aureus isolates obtained in the Northeast region of Brazil

Methicillin-resistant Staphylococcus aureus (MRSA) is a major agent of hospital infections worldwide. In Brazil, a multiresistant MRSA lineage (ST239-SCCmecIIIA), the so-called Brazilian epidemic clone (BEC), has predominated in all regions. However, an increase in nosocomial infections caused by non-multiresistant MRSA clones has recently been observed. In the present study, 45 clinical isolates of MRSA obtained from a university hospital located in Natal city, Brazil, were identified by standard laboratory methods and molecularly characterized using staphylococcal chromosome cassette mec (SCCmec) typing and pulsed-field gel electrophoresis. Antimicrobial susceptibility testing was carried out using CLSI methods. The MRSA isolates studied displayed a total of 8 different pulsed-field gel electrophoresis patterns (types A to H) with predominance (73%) of pattern A (BEC-related). However, MRSA harboring SCCmec type IV were also identified, 3 (7%) of which were genetically related to the pediatric clone--USA800 (ST5-SCCmecIV). In addition, we found a considerable genetic diversity within BEC isolates. MRSA displaying SCCmecIV are frequently susceptible to the majority of non-beta-lactam antibiotics. However, emergence of multiresistant variants of USA800 was detected.

Candida species distribution, genotyping and virulence factors of Candida albicans isolated from the oral cavity of kidney transplant recipients of two geographic regions of Brazil

BackgroundCandida albicans is a diploid yeast that in some circumstances may cause oral or oropharyngeal infections. This investigation aimed to study the prevalence of Candida spp. and to analyze the ABC genotypes of 76 clinical isolates of C. albicans obtained from the oral cavity of kidney transplant patients from two distinct geographic regions of Brazil.MethodsWe typed 48 strains with ABC genotyping and Microsatelitte using primer M13 and tested three virulence factors in vitro: phospholipase activity, morphogenesis and the ability to evade from polymorphonuclear neutrophils phagocytosis.ResultsC. albicans was the most prevalent species (86.4%), followed by C. tropicalis (4.5%). C. albicans genotype A was the most prevalent (58 isolates; 76.4%), followed by genotype C (15 isolates; 19.7%) and genotype B (3 isolates; 3.9%). When Microsatellite technique with primer M13 was applied, 80% of the isolates from the South were placed within the same cluster. The majority of Genotype C strains were grouped together within two different clusters. Genotype C was considered more resistant to PMNs attack than genotypes A and B. Strains isolated from the South of Brazil showed also better ability to combat PMNs phagocytosis.ConclusionsWe found a high rate of C. albicans genotype C strains isolated from the oral cavity of this group of patients. This study characterized oral C. albicans strains isolated from kidney transplant recipients and will contribute to a better understanding of the pathogenesis of oral candidiasis.

Evaluation of different methods for detecting methicillin resistance in Staphylococcus aureus isolates in a university hospital located in the Northeast of Brazil

Many methods have been described for the detection of methicillin-resistant Staphylococcus aureus (MRSA), but the heterogeneous expression of methicillin resistance affects the reliability of these methods. The aim of the present study was to evaluate some methods for detecting methicillin resistance in Staphylococcus aureus isolates in a university hospital located in the Northeast of Brazil. Among the isolates, 15 were methicillin-susceptible and 45 were methicillin-resistant, including low-level heterogeneous resistance strains. Both the 30 ηg-cefoxitin disk and PBP2a test had 100% sensibility/specificity and appear to be good options for the detection of MRSA in the clinical laboratory.

Carbohydrate assimilation profiles of Brazilian Candida dubliniensis isolates based on ID 32C system

The purpose of the present study was to evaluate the identification of 19 Brazilian C. dubliniensis based on the biochemical profile exhibited when tested by the commercial identification kit ID 32C (bioMerieux). Thirteen of the isolates were rigorously identified as C. dubliniensis and the remaining isolates (six) were considered as having a doubtful profile but the software also suggested that there was 83.6% of chances for them to be C. dubliniensis. As well as pointed by the literature the identification obtained by phenotypic tests should be considered presumptive for C. dubliniensis due to variability of this new species.

Attenuated total reflection Fourier transform-infrared (ATR-FTIR) spectroscopy as a new technology for discrimination between Cryptococcus neoformans and Cryptococcus gattii

Systemic fungal infections are among the most difficult diseases to manage in humans, especially when the recognition of the correct species is required for a precise and successful treatment. This is the case for Cryptococcus species and its genotypes, which are the main cause of meningitides in immunocompromised patients. Attenuated total reflection Fourier transform-infrared (ATR-FTIR) spectroscopy with discriminant analysis was employed to distinguish between the pathogenic fungal species Cryptococcus neoformans and Cryptococcus gattii by determining which wavenumber–absorbance/intensity relationships might reveal biochemical differences. Cryptococcus inactivated colonies were applied to an ATR crystal, and vibrational spectra were obtained in the ATR mode. Twenty-eight Cryptococcus isolates, fourteen C. neoformans and fourteen C. gattii were investigated. Spectral categories were analyzed using principal component analysis (PCA), successive projection algorithm (SPA) and genetic algorithm (GA) followed by linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA). Multivariate classification accuracy results were estimated based on sensitivity, specificity, positive (or precision) and negative predictive values, Youden index, and positive and negative likelihood ratios. Sensitivity for C. neoformans and C. gattii categories were 84.4% and 89.3%, respectively, using a QDA-LDA model with 17 wavenumbers with respect to their “fingerprints”. Compared to classical methods for differentiation of Cryptococcus species, this new technology could represent an alternative and innovative tool for faster and cheaper fungal identification for routine diagnostic laboratories.

Effect of the crude extract of Eugenia uniflora in morphogenesis and secretion of hydrolytic enzymes in Candida albicans from the oral cavity of kidney transplant recipients

BackgroundCandida albicans is a diploid yeast that in some circumstances may cause oral or oropharyngeal infections. Yeasts virulence factors contribute for both the maintenance of colonizing strains in addition to damage and cause tissue invasion, thus the establishment of infection occurs. The limited arsenal of antifungal drugs for the treatment of candidiasis turn the investigation of natural products mandatory for the discovery of new targets for antifungal drug development. Therefore, tropical countries emerge as important providers of natural products with potential antimicrobial activity. This study aimed to investigate morphogenesis and secretion of hydrolytic enzymes (phospholipase and proteinase) in the presence of the CE of Eugenia uniflora.MethodsThe isolates were tested for their ability to form hyphae in both solid and liquid media under three different conditions: YPD + 20% FBS, Spider medium and GlcNac and the ability to secrete phospholipase and proteinase in the presence of 2000 μg/mL of E. uniflora.ResultsThe CE of E. uniflora inhibited hypha formation in both liquid and solid media tested. It also impaired hydrolytic enzymes production.ConclusionsThis was the first study to describe the interaction of a natural product with the full expression of three different factors in C. albicans. E. uniflora may be an alternative therapeutic for oral candidiasis in the future.

Rhinocerebral zygomycosis in a diabetic patient

Rhinocerebral zygomycosis is the most frequent form of fungal infection caused by members of the Zygomycetes class. A fatal case of rhinocerebral zygomycosis caused by Rhizopus (oryzae) arrhizus with histopathological and mycological diagnosis is reported in a diabetic patient.

A 7-year survey of superficial and cutaneous mycoses in a public hospital in Natal, Northeast Brazil

In the present study, we determined the frequency of superficial and cutaneous mycoses and their etiologic agents during a 7-year period (2002–2008) in Natal, Brazil. A total of 1,717 specimens of skin, nail, and hair were collected from 1,382 patients with suspected mycoses lesions and were then subjected to direct microscopy and culture.