Abstract:SUMMARYThe basidiomycetous yeast Cryptococcus neoformans is an important fungal pathogen mainly in immunocompromised patients. In this study, 47 clinical isolates of C. neoformans from regions of São Paulo State were studied serologically by using the Crypto Check Iatron RM 304-K kit, their genetic diversity was estimated by PCR-fingerprinting with a microsatellite-specific sequence (GACA) 4 , RAPD with primer 6 (Amersham Pharmacia Biotech), PCR-restriction fragment length polymorphism (RFLP) analysis of the p… Show more
“…However, in Brazil, molecular epidemiology studies are still required to elucidate the distribution of molecular types in all five Brazilian regions. Despite this, the few epidemiological studies conducted in Brazil have demonstrated differences in the distribution of the genotypes of these species in these regions [17][18][19][20][21]36]. In Mato Grosso, this is the first report describing the molecular types circulating in the state capital of Cuiabá.…”
Section: Discussionmentioning
confidence: 88%
“…C. neoformans is the species with the largest number of isolates among species belonging to the Cryptococcus complex [5,[16][17][18]37]. Currently, an increasing number of cases of cryptococcosis have been reported in patients without HIV.…”
Section: Discussionmentioning
confidence: 99%
“…In Brazil, cryptococcosis caused by C. neoformans occurs in all regions of the country [5,[16][17][18][19][20]. However, C. gattii has emerged as a primary pathogen infecting immunocompetent individuals, particularly children, adolescents and young adults in northeastern Brazil, where cryptococcosis is characterized by high mortality rates [18,[20][21][22].…”
Introduction: Cryptococcosis is a systemic fungal infection that affects humans and animals, mainly due to Cryptococcus neoformans and Cryptococcus gattii. Following the epidemic of acquired immunodeficiency syndrome (AIDS), fungal infections by C. neoformans have become more common among immunocompromised patients. Cryptococcus gattii has primarily been isolated as a primary pathogen in healthy hosts and occurs endemically in northern and northeastern Brazil. We to perform genotypic characterization and determine the in vitro susceptibility profile to antifungal drugs of the Cryptococcus species complex isolated from HIV-positive and HIV-negative patients attended at university hospitals in Cuiabá, MT, in the Midwestern region of Brazil. Methodology: Micromorphological features, chemotyping with canavanine-glycine-bromothymol blue (CGB) agar and genotyping by URA5-RFLP were used to identify the species. The antifungal drugs tested were amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole. Minimum inhibitory concentrations (MICs) were determined according to the CLSI methodology M27-A3. Results: Analysis of samples yelded C. neoformans AFLP1/VNI (17/27, 63.0%) and C. gattii AFLP6/VGII (10/27, 37.0%). The MICs ranges for the antifungal drugs were: amphotericin B (0.5-1 mg/L), fluconazole (1-16 mg/L), flucytosine (1-16 mg/L), itraconazole (0.25-0.12 mg/L) and voriconazole (0.06-0.5 mg/L). Isolates of C. neoformans AFLP1/VNI were predominant in patients with HIV/AIDS, and C. gattii VGII in HIV-negative patients. The genotypes identified were susceptible to the antifungal drugs tested. Conclusion: It is worth emphasizing that AFLP6/VGII is a predominant genotype affecting HIV-negative individuals in Cuiabá. These findings serve as a guide concerning the molecular epidemiology of C. neoformans and C. gattii in the State of Mato Grosso.
“…However, in Brazil, molecular epidemiology studies are still required to elucidate the distribution of molecular types in all five Brazilian regions. Despite this, the few epidemiological studies conducted in Brazil have demonstrated differences in the distribution of the genotypes of these species in these regions [17][18][19][20][21]36]. In Mato Grosso, this is the first report describing the molecular types circulating in the state capital of Cuiabá.…”
Section: Discussionmentioning
confidence: 88%
“…C. neoformans is the species with the largest number of isolates among species belonging to the Cryptococcus complex [5,[16][17][18]37]. Currently, an increasing number of cases of cryptococcosis have been reported in patients without HIV.…”
Section: Discussionmentioning
confidence: 99%
“…In Brazil, cryptococcosis caused by C. neoformans occurs in all regions of the country [5,[16][17][18][19][20]. However, C. gattii has emerged as a primary pathogen infecting immunocompetent individuals, particularly children, adolescents and young adults in northeastern Brazil, where cryptococcosis is characterized by high mortality rates [18,[20][21][22].…”
Introduction: Cryptococcosis is a systemic fungal infection that affects humans and animals, mainly due to Cryptococcus neoformans and Cryptococcus gattii. Following the epidemic of acquired immunodeficiency syndrome (AIDS), fungal infections by C. neoformans have become more common among immunocompromised patients. Cryptococcus gattii has primarily been isolated as a primary pathogen in healthy hosts and occurs endemically in northern and northeastern Brazil. We to perform genotypic characterization and determine the in vitro susceptibility profile to antifungal drugs of the Cryptococcus species complex isolated from HIV-positive and HIV-negative patients attended at university hospitals in Cuiabá, MT, in the Midwestern region of Brazil. Methodology: Micromorphological features, chemotyping with canavanine-glycine-bromothymol blue (CGB) agar and genotyping by URA5-RFLP were used to identify the species. The antifungal drugs tested were amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole. Minimum inhibitory concentrations (MICs) were determined according to the CLSI methodology M27-A3. Results: Analysis of samples yelded C. neoformans AFLP1/VNI (17/27, 63.0%) and C. gattii AFLP6/VGII (10/27, 37.0%). The MICs ranges for the antifungal drugs were: amphotericin B (0.5-1 mg/L), fluconazole (1-16 mg/L), flucytosine (1-16 mg/L), itraconazole (0.25-0.12 mg/L) and voriconazole (0.06-0.5 mg/L). Isolates of C. neoformans AFLP1/VNI were predominant in patients with HIV/AIDS, and C. gattii VGII in HIV-negative patients. The genotypes identified were susceptible to the antifungal drugs tested. Conclusion: It is worth emphasizing that AFLP6/VGII is a predominant genotype affecting HIV-negative individuals in Cuiabá. These findings serve as a guide concerning the molecular epidemiology of C. neoformans and C. gattii in the State of Mato Grosso.
“…Only a few Brazilian studies have investigated the molecular types of environmental Cryptococcus 1,2,3,22,24,28,35 . The distribution of molecular types using the specific primer M13 shows the predominance of VNI (serotype A) in surveyed regions.…”
mentioning
confidence: 99%
“…It is worthwhile to note the presence of VNIV (serotype D) in eucalypt trees in the deep south of Brazil, a subtropical region 2,28 , and of VGI (C. gattii serotype B) in excreta of psittaciformes in the same region 1 . Study of clinical isolates in São Paulo showed that all of them were serotype A and the majority belonged to the molecular type VNI 22 .…”
SUMMARYCryptococcus neoformans is the major cause of fungal meningitis, a potentially lethal mycosis. Bird excreta can be considered a significant environmental reservoir of this species in urban areas, thirty-three samples of pigeon excreta were collected within the city of Vitoria, Brazil. Cryptococcus neoformans was isolated and identified using standard biochemical assays in ten samples. PCR amplification with primer M13 and orotidine monophosphate pyrophosphorylase (URA5) gene-restriction fragment length polymorphism (RFLP) analysis discerned serotypes and genotypes within this species. All isolates were serotype A (C. neoformans var. grubii) and genotype VNI. The two alternative alleles a and a at the mating type locus were determined by PCR amplification and mating assays performed on V8 medium. All isolates were MAT a mating type but only 50% were able to mate in vitro with the opposite mating type MAT a tester strains (JEC20, KN99a and Bt63). This study adds information on the ecology and molecular characterization of C. neoformans in the Southeast region of Brazil.
To determine the profiles of susceptibility to antifungal and the genotypes of clinical isolates of Cryptococcus in Bahia, Brazil, 62 isolates were collected from cases of meningitis in the period from 2006 to 2010. Their susceptibilities to fluconazole, itraconazole, amphotericin B and 5-flucytosine were determined by the broth microdilution technique described by the Clinical and Laboratory Standards Institute and genotyping of the URA5 gene was accomplished by restriction fragment length polymorphism. C. neoformans accounted for 79% of the identified yeast and C. gattii represented the remaining 21%. Evaluation of the genotypes determined that 100% of the C. gattii isolates belong to the VGII genotype, and 98% of the C. neoformans isolates belong to the VNI genotype. Determination of susceptibility revealed isolates resistant to fluconazole (4.8%), 5-flucytosine (1.6%) and amphotericin B (3.2%); the stratification of sensitivity results for each species showed significant differences in susceptibility to azoles. This study is the first to describe the susceptibility profiles of molecular and clinical isolates of Cryptococcus in Bahia, Brazil. The high percentage of C. gattii isolates belonging to the VGII genotype and its lower susceptibility to antifungal agents highlight the importance of knowing which species are involved in cryptococcal infections in northeastern Brazil.
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