In order to study the infectious agents causing human disseminated cryptococcosis in the state of Pará, North Brazil, 56 isolates of Cryptococcusspp. (54 isolated from cerebral spinal fluid and two from blood cultures) from 43 cases diagnosed between 2003-2007 were analysed. The species were determined through morphological and physiological tests and genotypes were determined by URA5-RFLP and PCR-fingerprinting (wild-type phage M13). The following species and genotypes were identified: Cryptococcus neoformans VNI (28/56, 50%), Cryptococcus gattii VGII (25/56, 44.64%) and C. gattii VGI (3/56, 5.26%). The genotype VNI occurred in 12 out of 14 HIV-positive adults, whereas the genotype VGII occurred in 11 out of 21 HIV-negative adults (p < 0.02, OR = 6.6 IC95% 0.98-56.0). All patients less than 12 years old were HIV negative and six cases were caused by the VGII genotype, one by the VGI and one by VNI. Therefore, endemic primary mycosis in HIV-negative individuals, including an unexpectedly high number of children, caused by the VGII genotype deserves further study and suggests the need for surveillance on cryptococcal infection in the state of Pará, Eastern Amazon.

The molecular types of 443 Brazilian isolates of Cryptococcus neoformans and Cryptococcus gattii were analyzed to determine their geographic distribution within Brazil and their underlying host conditions. The following data, imported from previous epidemiological studies as well as two culture collections, were analyzed for: place of isolation, source (clinical or environmental), host risk factors, species, serotype, mating type, and molecular type. Molecular typing by PCR-fingerprinting using primers for the minisatellite-specific core sequence of the wild-type phage M13 or microsatellites [(GACA)4, (GTG)5], restriction fragment length polymorphism of URA5 gene analysis, and/or amplified fragment length polymorphism (AFLP) identified eight major genotypes: VNI/AFLP1, VNII/AFLP1A, VNIII/AFLP2, and VNIV/AFLP3 for C. neoformans, and VGI/AFLP4, VGII/AFLP6, VGIII/AFLP5, and VGIV/AFLP7 for C. gattii. The most common molecular type found in Brazil was VNI (64%), followed by VGII (21%), VNII (5%), VGIII (4%), VGI and VNIV (3% each), and VNIII (< 1%). Primary cryptococcosis caused by the molecular type VGII (serotype B, MAT) prevails in immunocompetent hosts in the North and Northeast regions, disclosing an endemic regional pattern for this specific molecular type in the Northern Brazil.

Introducción. En el complejo Cryptococcus neoformans se reconocen dos especies y cinco serotipos: C. neoformans (var. grubii, serotipo A; var. neoformans, serotipo D y un híbrido, serotipo AD) y C. gattii (serotipos B y C). La pareja sexual a y alfa es controlada por un solo locus, y la pareja sexual alfa es la más prevalente en los serotipos A y D, y es convencionalmente determinada mediante reacción en cadena de la polimerasa. Objetivo. Evaluar la habilidad de aislamientos colombianos de C. neoformans para aparearse in vitro con aislamientos control de la pareja sexual opuesta. Materiales y métodos. Treinta y tres aislamientos clínicos de C. neoformans var. grubii serotipo A, 4 de la var. neoformans serotipo D, todos pareja sexual alfa, y 16 aislamientos clínicos de C. gattii, 13 serotipo B (pareja sexual a) y 3 serotipo C (pareja sexual alfa), se mezclaron, en agar jugo V8 modificado, con cepas control para determinar la pareja sexual in vitro. Resultados. Los estudios de apareamiento mostraron que 9 de 33 (27,3%) aislamientos serotipo A y 6 de 13 (46,2%) aislamientos serotipo B tuvieron la capacidad de aparearse con las cepas control. Todos los aislamientos del serotipo A que presentaron apareamiento eran pareja sexual alfa y los del serotipo B eran pareja sexual a. Microscópicamente se observaron conexiones en gancho, basidias y basidiosporas, estructuras que establecieron que se había realizado el proceso de apareamiento. Conclusión. Este acercamiento provee por primera vez la capacidad de que los aislamientos colombianos de C. neoformans se apareen in vitro con cepas control lo cual tiene importancia en el estudio de la diseminación del hongo.

Introduction. Within the Cryptococcus neoformans species complex, two species and five serotypes are recognized: C. neoformans (var. grubii, serotype A; var. neoformans, serotype D and a hybrid, serotype AD) and C. gattii (serotypes B and C). Mating types a and alpha are designated by a single locus, with the mating type alpha being most prevalent in serotype A and D strains. Objective. To evaluate the ability of Colombian isolates of the C. neoformans species complex to mate in vitro with tester strains of the opposite mating type. Materials and methods. Fifty three clinical isolates were included in this study, 33 C. neoformans var. grubii serotype A, 4 C. neoformans var. neoformans serotype D, all mating type alpha, and 16 C. gattii, 13 serotype B (mating type a) and 3 serotype C (mating type alpha), were mixed on V8 juice agar, using a modified method, with the appropriate tester strains to determine the mating types in vitro. Results. Mating studies revealed that 9 of 33 (27.3%) serotype A isolates and 6 of 13 (46.2%) serotype B isolates were able to mate. Clamp connections and basidia with basidiospores were observed microscopically, indicating that the mating process had occurred. All mating competent serotype A strains were mating type alpha and the serotype B mating competent strains were mating type a. Conclusion. This is the first report of the determination of the mating ability of Colombian Cryptococcus neoformans isolates to mate in vitro with appropriate tester strains, which is of great importance to study the propagation of the fungus around the globe.