title: Antifungal susceptibility, enzymatic activity, PCR-fingerprinting and ITS sequencing of environmental Cryptococcus laurentii isolates from Uberaba, Minas Gerais, Brazil.
abstract: Cryptococcus laurentii has been classically considered a saprophytic species, although several cases of human infection have been already reported. This study aimed to evaluate the phospholipase, proteinase and hemolysins activity, the antifungal susceptibility profile, the genetic variability by M13 and (GACA)(4) fingerprinting and the internal transcribe spacer (ITS) sequencing of 38 C. laurentii isolates recovered from captive bird droppings and surrounding hospital areas. All of them exhibited phospholipase activity, while the hemolytic activity was evidenced in 34 (89.4%) isolates. None of them exhibited proteinase activity. Twenty-seven isolates (71.1%) presented susceptibility dose dependent to fluconazole. Most isolates (94.7%) were susceptible to voriconazole, while one (2.65%) was resistant to this drug. Twenty-one (55.3%) isolates showed reduced susceptibility to itraconazole while nine (23.7%) were resistant. Three (7.9%) and five (13.1%) isolates exhibited resistance to ketoconazole and amphotericin B, respectively. Most C. laurentii fingerprinting obtained with M13 and (GACA)(4) showed high heterogeneity. By using the two primers, seven (18.4%) isolates grouped as A (CL2, CL7, and CL8), B (CL35, CL38) and C (CL29, CL30) with 100% similarity. Different from most variable surrounding hospital isolates, all but one of the pet shops strains clustered with the two primers, although they had been recovered from different neighborhoods. All isolates were identified as C. laurentii phylogenetic group I by ITS sequencing. Thus, the presence of virulence factors, a decreased antifungal susceptibility and a heterogeneous molecular pattern of the C. laurentii isolates here described suggests this species can be a potential pathogen in the context of the immunocompromised population.
authors: Department of Infectious and Parasitic Diseases, Triangulo Mineiro Federal University, Postal Code 118, Uberaba, MG, 38001-170, Brazil. kennio_p@hotmail.com
authors: Kennio
authors: K
authors: Ferreira-Paim
authors: Leonardo
authors: L
authors: Andrade-Silva
authors: Delio Jose
authors: DJ
authors: Mora
authors: Eliane
authors: E
authors: Lages-Silva
authors: André Luiz
authors: AL
authors: Pedrosa
authors: Paulo Roberto
authors: PR
authors: da Silva
authors: Anderson Assunção
authors: AA
authors: Andrade
authors: Mario León
authors: ML
authors: Silva-Vergara
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology*; D001717:Birds; D001938:Brazil; D016000:Cluster Analysis; D003454:Cryptococcus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D016172:DNA Fingerprinting; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D004783:Environmental Microbiology; D004798:Enzymes / Q000378:metabolism*; D005243:Feces / Q000382:microbiology; D014644:Genetic Variation; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D058889:Molecular Typing; D016533:Mycological Typing Techniques; D010802:Phylogeny; D017422:Sequence Analysis, DNA
chemical_list: D000935:Antifungal Agents; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D004798:Enzymes