Florencia Rojas

Prevalence of Malassezia species in pityriasis versicolor lesions in northeast Argentina

BACKGROUND Malassezia species normally colonize the skin but they can change their saprophytic state and invade the stratum corneum as pathogens. AIMS To determine the prevalence of Malassezia species isolated from patients with pityriasis versicolor (PV) and to analyse their distribution according to the location of the lesion on the body. METHODS This study included 218 patients with PV and positive Malassezia cultures who resided in the city of Resistencia, a subtropical area located in northeast Argentina. Age, gender, and the body site of lesions were recorded. Strains were identified by PCR-RFLP. RESULTS Malassezia sympodialis (37.7%) and Malassezia globosa (37.2%) were the most prevalent species isolated alone or in association with other Malassezia species in 82% of the patients. Malassezia furfur (21.3%) was the third most common species, followed by Malassezia slooffiae (1.7%), and Malassezia restricta (1.3%), which was found only in combination with M. globosa and M. sympodialis. Malassezia dermatis (0.4%) and Malassezia pachydermatis (0.4%) were each isolated once. None of the species affected a body site with statistical significance. Significant difference between genders according to age was found only in the 31-40-year-age group. CONCLUSIONS This study suggests that M. sympodialis and M. globosa represent the main species implicated in the pathogenicity of PV. M. furfur appears to be the third agent of importance in this geographical area. Statistical analyses showed none of the species was particularly associated with any one of the body sites.

Fluconazole and Itraconazole Resistance of Yeasts Isolated from the Bloodstream and Catheters of Hospitalized Pediatric Patients

Background: In critically ill children, Candida species and other yeasts appear as an important nosocomial pathogen. The emerging fungal pathogens are usually less susceptible to azole compounds, and the management of such infections could be problematic. Methods: 6,065 bloodstream cultures and 627 catheters from intensive care units and hospitalized oncology pediatric patients were studied. Antifungal susceptibility testing of isolates was performed according to the reference broth microdilution method describedby the National Committee for Clinical Laboratory Standards (M27-A). Results: We found a low activity of fluconazole (FCZ) and itraconazole (ITZ) against Candida glabrata, C. albicans, C. tropicalis and C. haemulonii. Resistant strains to FCZ and ITZ were detected. Conclusion: Results reveal the emergence of antifungal-resistant species and a change in the predominant role of C. albicans as a cause of candidemia in hospitalized children.

Yeasts species distribution in Neonatal Intensive Care Units in northeast Argentina

The aim of this study was to determine the distribution and antifungal susceptibility profile of yeast species isolated from neonates in Neonatal Intensive Care Units (NICU) in northeast of Argentina. With this purpose 92 strains isolated from 25 blood stream cultures, 20 venous catheters, 23 suprapubic aspirations and 24 rectal swabs were studied. Candida albicans and C. parapsilosis appeared with similar frequencies (36%) in blood stream isolates. Candida parapsilosis (50%) was the most frequent catheters colonizer and C. tropicalis (54.2%) was the most frequent yeast associated with gastrointestinal tract colonization. Candida krusei, C. glabrata and Trichosporon cutaneum appeared with a very low frequency. A high rate of susceptibility to amphotericin B, fluconazole, and itraconazole was observed.

In vitro antifungal activity of topical and systemic antifungal drugs against Malassezia species

The strict nutritional requirements of Malassezia species make it difficult to test the antifungal susceptibility. Treatments of the chronic and recurrent infections associated with Malassezia spp. are usually ineffective. The objective of this study was to obtain in vitro susceptibility profile of 76 clinical isolates of Malassezia species against 16 antifungal drugs used for topical or systemic treatment. Isolates were identified by restriction fragment length polymorphism. Minimal inhibitory concentrations (MIC) were obtained by a modified microdilution method based on the Clinical Laboratory Standards Institute reference document M27‐A3. The modifications allowed a good growth of all tested species. High in vitro antifungal activity of most tested drugs was observed, especially triazole derivatives, except for fluconazole which presented the highest MICs and widest range of concentrations. Ketoconazole and itraconazole demonstrated a great activity. Higher MICs values were obtained with Malassezia furfur indicating a low susceptibility to most of the antifungal agents tested. Malassezia sympodialis and Malassezia pachydermatis were found to be more‐susceptible species than M. furfur, Malassezia globosa, Malassezia slooffiae and Malassezia restricta. Topical substances were also active but provide higher MICs than the compounds for systemic use. The differences observed in the antifungals activity and interspecies variability demonstrated the importance to studying the susceptibility profile of each species to obtain reliable information for defining an effective treatment regimen.

Aspergillus terreus complex: an emergent opportunistic agent of Onychomycosis.

The incidence of onychomycosis due to non‐dermatophyte moulds (NDM) is increasing. Aspergillus terreus is relatively undocumented as an agent of this fungal infection. The aim of this work is to show the prevalence of onychomycosis caused by A. terreus and to describe its clinical features. Nail samples were collected for microscopic examination and culturing in selective media. All cases of onychomycosis due to NDM were confirmed by a second sample. Aspergillus terreus isolates were identified through their morphological characteristics and using molecular methods. A total of 2485 samples were obtained. Positive cultures were obtained in 1639 samples. From 124 NDM confirmed cultures, 23 were identified as A. terreus (18.5%). Superficial white onychomycosis was the most frequent clinical pattern. A high percentage was found in fingernails. The prevalence of A. terreus in this study considerably exceeded the percentages reported by other authors. Onychomycosis due to A. terreus presents similar clinical patterns to those caused by dermatophytes, but is difficult to eradicate and is associated with less predictable treatment outcomes. Better knowledge of the aetiology of A. terreus may be important for accomplishing more accurate and effective treatment.

Native trees of the Northeast Argentine: Natural hosts of the Cryptococcus neoformans–Cryptococcus gattii species complex

BACKGROUND In Argentina, information about epidemiology and environmental distribution of Cryptococcus is scarce. The city of Resistencia borders with Brazil and Paraguay where this fungus is endemic. All these supported the need to investigate the ecology of the genus and the epidemiology of cryptococcosis in this area. AIMS The aim was to investigate the presence of species of Cryptococcus neoformans-Cryptococcus gattii complex and their genotypes in trees of the city of Resistencia. METHODS One hundred and five trees were sampled by swabbing technique. The isolates were identified using conventional and commercial methods and genotyped by PCR-RFLP (Restriction Fragment Length Polymorphism). RESULTS Cryptococcus was found in 7 out of the total trees. 6 out of 7 Cryptococcus isolates were identified as C. neoformans and one as C. gattii. C. gattii was isolated from Grevillea robusta. C. neoformans strains were isolated from Tabebuia avellanedae and Peltophorum dubium. Genotyping showed that all C. neoformans belonged to the VNI type and C. gattii belonged to the VGI type. CONCLUSIONS This represents the first study on the ecology of Cryptococcus spp. associated to trees from northeastern Argentina, and the first report describing Grevillea robusta as a host of members of this fungal genus. Another finding is the isolation of C. neoformans from Tabebuia avellanedae and Peltophorum dubium, both tree species native to northeastern Argentina.

Antifungal susceptibility of Malassezia furfur, Malassezia sympodialis, and Malassezia globosa to azole drugs and amphotericin B evaluated using a broth microdilution method

We studied the in vitro activity of fluconazole (FCZ), ketoconazole (KTZ), miconazole (MCZ), voriconazole (VCZ), itraconazole (ITZ) and amphotericin B (AMB) against the three major pathogenic Malassezia species, M. globosa, M. sympodialis, and M. furfur. Antifungal susceptibilities were determined using the broth microdilution method in accordance with Clinical and Laboratory Standards Institute reference document M27-A3. To support lipid-dependent yeast development, glucose, peptone, ox bile, malt extract, glycerol, and Tween supplements were added to Roswell Park Memorial Institute RPMI 1640 medium. The supplemented medium allowed good growth of all three species studied. The minimal inhibitory concentrations (MICs) were recorded after 72 h of incubation at 32ºC. The three species showed different susceptibility profiles for the drugs tested. Malassezia sympodialis was the most susceptible and M. furfur the least susceptible species. KTZ, ITZ, and VCZ were the most active drugs, showing low variability among isolates of the same species. FCZ, MCZ, and AMB showed high MICs and wide MIC ranges. Differences observed emphasize the need to accurately identify and evaluate antifungal susceptibility of Malassezia species. Further investigations and collaborative studies are essential for correlating in vitro results with clinical outcomes since the existing limited data do not allow definitive conclusions.

Etiology of fungaemia and catheter colonisation in Argentinean paediatric patients.

Yeast strains obtained from blood cultures and catheters from intensive care units (ICU) and hospitalised oncology paediatrics were studied. Yeast were the first cause of catheter colonisation (51/627), and the third cause of bloodstream infection (44/6065). In catheter, the most frequent species were Candida albicans (34%), C. parapsilosis (27.7%) and C. tropicalis (15%). In blood, C. albicans (40.8%), C. parapsilosis (26.6%), C. tropicalis (15%). Malassezia furfur and Malassezia sympodialis were isolated from catheters from ICU patients. All isolates were susceptible to amphotericin B, 88.8% to itraconazole and 91.9% to fluconazole. Candida albicans and C. tropicalis strains resistant to fluconazole and itraconazol were detected. These results reveal a change in the predominant role of C. albicans as cause of candidemia in hospitalised children and the emergence of antifungal resistant species. These variations emphasise the importance of performing a permanent surveillance to observe and assess them.

Antifungal susceptibility testing of Malassezia yeast: comparison of two different methodologies.

All Malassezia species are lipophilic; thus, modifications are required in susceptibility testing methods to ensure their growth. Antifungal susceptibility of Malassezia species using agar and broth dilution methods has been studied. Currently, few tests using disc diffusion methods are being performed. The aim was to evaluate the in vitro susceptibility of Malassezia yeast against antifungal agents using broth microdilution and disc diffusion methods, then to compare both methodologies. Fifty Malassezia isolates were studied. Microdilution method was performed as described in reference document and agar diffusion test was performed using antifungal tablets and discs. To support growth, culture media were supplemented. To correlate methods, linear regression analysis and categorical agreement was determined. The strongest linear association was observed for fluconazole and miconazole. The highest agreement between both methods was observed for itraconazole and voriconazole and the lowest for amphotericin B and fluconazole. Although modifications made to disc diffusion method allowed to obtain susceptibility data for Malassezia yeast, variables cannot be associated through a linear correlation model, indicating that inhibition zone values cannot predict MIC value. According to the results, disc diffusion assay may not represent an alternative to determine antifungal susceptibility of Malassezia yeast.

Genotipos y epidemiología de aislamientos clínicos de Cryptococcus neoformans en Corrientes, Argentina

Fil: Cattana, Maria Emilia. Universidad Nacional del Nordeste. Instituto de Medicina Regional; Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas; Argentina