Genotypes of Cryptococcus neoformans and Cryptococcus gattii as agents of endemic cryptococcosis in Teresina, Piauí (northeastern Brazil)

Throughout Brazil, Cryptococcus neoformans is the cause of cryptococcosis, whereas Cryptococcus gattii is endemic to the northern and northeastern states. In this study, the molecular types of 63 cryptococcal isolates recovered from the cerebrospinal fluid of meningitis patients diagnosed between 2008-2010 in Teresina, Piauí, Brazil, were analysed. Out of the 63 patients, 37 (58.7 percent) were human immunodeficiency virus (HIV)-positive and 26 (41.3 percent) were HIV-negative. URA5-restriction fragment length polymorphism analysis identified 37/63 (58.7 percent) isolates as the C. neoformans VNI genotype, predominantly in HIV-positive patients (32/37, 86.5 percent), and 24/63 (38.1 percent) as the C. gattii VGII genotype, mostly in HIV-negative patients (21/26, 80.8 percent). The occurrence of C. gattii VGII in six apparently healthy children and in seven adolescents/young adults in this region reaffirms the endemic occurrence of C. gattii VGII-induced primary cryptococcosis and early cryptococcal infection. Lethality occurred in 18/37 (48.6 percent) of the HIV-positive subjects and in 13/26 (50 percent) of the HIV-negative patients. Our results provide new information on the molecular epidemiology of C. neoformans and C. gattii in Brazilian endemic areas.

Primary endemic Cryptococcosis gattii by molecular type VGII in the state of Pará, Brazil

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 percent), Cryptococcus gattii VGII (25/56, 44.64 percent) and C. gattii VGI (3/56, 5.26 percent). 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 percent 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.

Isolamento e identificação de Coccidioides immitis de amostras de solo relacionadas a surtos de Coccidioidomicose / Isolation and identification of Coccidioides immitis from soil samples related to outbreaks of coccidioidomycosis

A coccidioidomicose (CM) é micose sistêmica endêmica no semi-árido do nordeste brasileiro causada pelo fungo dimórfico geofílico Coccidioides posadasii [=C.immitis (non-CA)].No Brasil a principal característica epidemiológica relacionada a surtos da CM aguda é o distúrbio ambiental provocado por caçadores de tatus que, ao desentocarem o animal de seu habitat natural, revolvem solo contaminado e se infectam ao inalar propágulos (artroconídios) do fungo dispersos na poeira do ar, desencadeando o processo de infecção e doença.O presente trabalho teve como objetivos(i)Isolar e identificar por metodologias clássicas agentes de micoses sistêmicas em amostras ambientais relacionadas a surtos de CM;(ii) Avaliar a eficiência de protocolos para extração de DNA a partir das amostras ambientais utilizando duas metodologias distintas:a)criofratura com nitrogênio líquido e b)kit de extração comercial Fast DNA® SPIN® Kit For Soil Q-BIOgene;(iii)Desenvolver iniciadores específicos para a detecção de C.posadasii utilizando recursos de bioinformática;(iv)Avaliar e validar os iniciadores específicos,utilizando-os na detecção de C. posadasii em DNAs de amostras de solo.Foram analisadas por estas metodologias amostras ambientais de dois sítios suspeitos de contaminação por C.posadasii.De 10 amostras de solo coletadas em Elesbão Veloso,3(30 por cento) foram positivas para C.posadasii por inoculação animal,2(20 por cento)foram positivas pela PCR direta e 10 (100 por cento) foram positivas no rastreamento com a técnica de PCR semi-nested. De 23 amostras de solo coletadas em Caridade do Piauí,3(13,3 por cento) foram positivas para C. posadasii por inoculação animal,6(42,8 por cento) de 14 amostras foram positivas pela PCR direta e todas estas 14 amostras (100 por cento) foram positivas no rastreamento com a PCR semi-nested. De 23 amostras de solo coletadas em Caridade do Piauí,3(13,3 por cento) foram positicas para C. immitis non-CA por inoculação animal,6 (42,8 por cento) de amostra...