Made available in DSpace on 2016-04-28T20:16:31Z (GMT). No. of bitstreams: 1 2008 - Maria Cristina Forbes Santos Bustamante.pdf: 1772291 bytes, checksum: 84cc7e884259a3fc0a8ca095cc88b7df (MD5) Previous issue date: 2009-12-19 / Funda??o Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / The knowledge about epidemiology of American Cutaneous Leishmaniasis - ACL still has gaps. The influences of environmental and peridomestic characteristics over its occurrence are not well defined. This study aimed to investigate the asymptomatic occurrence of Leishmania (Viannia) braziliensis in humans and dogs, and to evaluate the natural infection of phlebotomine vectors at Engenheiro Paulo de Frontin, a disease-free of autochthonous ACL municipality, part of an endemic region. An epidemiological inquiry was developed from march/2006 to december/2007 with the support of the Family Health Program of the Municipality Health Secretary Office in the five sanitary areas. Fifty families participated in the study, yielding a total of 95 volunteers who were exposed or not to known risk factors. Procedures involved structured interviews, Montenegro skin Test (IDRM), serology done by Enzime Linked Immunossorbent Assay (ELISA), and Indirect Immunofluorescence (IIF). All the volunteers had their dogs examined by the serology procedures and including a clinical exam. The existing environment was reported and phlebotomine sand flies were captured in the peridomestic environment to have all the present species identified. Infection by L. (V.) braziliensis was also determined by using polymerase chain reaction (PCR). Observed frequencies for positive results in humans were: 9%, 6.4% and 13.8% for IDRM, ELISA and IFI, respectively. Of 39 dogs examined, 5.1% were positive for ELISA and 23% for IFI, and from the seven species of phlebotomine sand flies captured, Lutzomyia migonei (59%) and L. intermedia (20.9%) were the most frequent. Natural infection rate of phlebotomine vectors were 5% or less, by PCR. The absence of active cases and the low percentage of positive tests found in humans and dogs, and the non-matching results, lead to the suggestion that there is no extra forest transmission of L. (V.) braziliensis in the county, despite of the high natural infection rate of phlebotomines sand flies. Large areas of preserved forests scattered around 52% of the municipality appears to be the only and main difference from the surrounding municipality, thus allowing maintenance of the ecological niches of the phlebotomine sand flies and reducing the pressure for adaptive changes. / O conhecimento acerca da epidemiologia da Leishmaniose Tegumentar Americana (LTA) ainda possui lacunas. As influ?ncias de caracter?sticas ambientais e peridom?sticas sobre sua ocorr?ncia n?o est?o bem definidas. Este estudo buscou investigar a poss?vel ocorr?ncia de infec??o assintom?tica por Leishmania (Viannia) braziliensis em humanos e c?es e a infec??o natural dos flebotom?neos vetores no munic?pio de Engenheiro Paulo de Frontin, indene para LTA aut?ctone e circundado por munic?pios end?micos. Foi realizado um inqu?rito epidemiol?gico entre mar?o de 2006 e dezembro de 2007, sendo investigadas as cinco ?reas de sa?de do munic?pio, com apoio do Programa de Sa?de da Fam?lia da Secretaria Municipal de Sa?de. Participaram do estudo cinq?enta fam?lias expostas ou n?o aos fatores de risco conhecidos, totalizando noventa e cinco volunt?rios. Entrevistas estruturadas, Intradermorrea??o de Montenegro (IDRM) e sorologia para leishmaniose, por ELISA e por Imunofluoresc?ncia Indireta (IFI) foram realizadas nos volunt?rios, cujos c?es foram tamb?m submetidos a exame cl?nico e sorologia pelos mesmos m?todos. O ambiente circundante foi descrito e flebotom?neos foram capturados nos peridomic?lios para identifica??o das esp?cies presentes e de infec??o por L. (V.) braziliensis pela t?cnica de rea??o em cadeia de polimerase (RCP). As freq??ncias encontradas de positividade nos testes em humanos foram de: 9%, 6,4% e 13,8% para IDRM, ELISA e IFI, respectivamente. Dos trinta e nove c?es examinados 5,1%, foram positivos ao ELISA e 23% ? IFI. Das sete esp?cies de flebotom?neos capturadas, predominaram Lutzomyia migonei (59%) e L. intermedia (20,9%). A taxa de infec??o natural dos flebotom?neos foi de at? 5% ? RCP. A aus?ncia de casos ativos e as baixas positividades verificadas nos testes em humanos e em c?es e a discord?ncia entre os resultados levam a supor que n?o ocorre transmiss?o extraflorestal de L. (V.) braziliensis no munic?pio, a despeito da alta taxa de infec??o natural dos flebotom?neos. Grandes ?reas de mata preservada dispersas pelo territ?rio (52% do munic?pio) parecem ser o ?nico diferencial em rela??o aos munic?pios circundantes, permitindo a manuten??o dos nichos ecol?gicos dos flebotom?neos e reduzindo a press?o por mudan?as adaptativas.

Cryptococcus neoformans species complex são fungos oportunistas que desencadeiam meningite, tanto em pacientes imunocompetentes como em indivíduos hígidos. Este trabalho teve como objetivo estudar as espécies circulantes de Cryptococcus neoformans species complex isolados no Estado de Sergipe tanto a partir do LCR (líquido céfalo raqueanao) de pacientes com meningite infecciosa como de diversos substratos ambientais. Foram utilizadas amostras do LCR que deram entrada no IPH/HEMOLACEN/SE no período de 2001 a 2007. Foram realizadas 104 coletas ambientais dentro do Estado de Sergipe, das quais 55 foram obtidas a partir de excretas de aves e quatro de amostras de fezes de morcegos coletadas em cavernas. As demais foram obtidas de materiais de origem vegetal, perfazendo 45 amostras: 21 de Eucalyptus spp., oito de material de troncos vegetais em decomposição, 10 de restos vegetais coletados em duas praças e seis de Anacardium occidentale L. (cajueiro). O líquor foi examinado por microscopia direta e cultivado a 25ºC e a 37ºC. No período de 2005 a 2007 foram obtidos 141 isolados ambientais e 35 isolados clínicos de Cryptococcus neoformans species complex. Foram determinadas inferências ecológicas e epidemiológicas dos isolados obtidos além de realização de provas para identificação presuntiva fenotípica em meioNíger e, das espécies em meio CGB, teste de sensibilidade frente aosantifúngicos, caracterização enzimática de interesse biotipar e/ou marcador de linhagens, determinação de mating type sexual utilizando PCR e PCR Fingerprintig com iniciador M13. Nas linhagens de Cryptococcus spp. isoladas de líquor de pacientes com meningite infecciosa no Estado de Sergipe prevaleceu a espécie C. neoformans. Em relação às linhagens ambientais, somente uma amostra, dentre 140, foi identifcada como C. gattii. Todas as amostras oriundas de excretas de aves foram positivas no isolamento da levedura. Os isolados clínicos de C. neoformans e C. gattii oriundos de pacientes com meningite criptocócica no Estado de Sergipe apresentaram valores altos de concentração inibitória mínima (CIM) para os agentes antifúngicos testados, o que simboliza alta taxa de resistência, principalmente à Anfotericina B e ao Fluconazol. Em relação ao mating type sexual foi evidenciando que a maioria das linhagens clínicas foi identificada como MAT, confirmando o caráter de virulência que esta levedura desenvolve. Entretanto, no isolados ambientais o MATa foi mais prevalente, evidenciando o caráter menos virulento destas linhagens frente às de origem clínica. Cryptococcus neoformans foi a única espécie amplificada por PCR fingerprinting. Houve presença, dentre as amostras locais, de três perfis genéticos distintos epertencentes a uma mesma espécie. _________________________________________________________________________________________ ABSTRACT: Cryptococcus neoformans species complex are opportunistic fungi that cause meningitis in both immunocompetent patients and in healthy individuals. This work aimed to study the species of circulating Cryptococcus neoformans species complex isolated from northeastern Brazil from both the CSF (cerebrospinal fluid) of patients with infectious meningitis as various environmental substrates. Samples of CSF were received at IPH/HEMOLACEN/SE in the period 2001 to 2007. Were carried out 104 environmental samples within the state of Sergipe, of which 55 were obtained from the feces of birds and four stool samples collected from bat caves. The other samples were obtained from plant materials, totaling 45 samples: 21 from Eucalyptus spp., eight from material of decomposing trunks of plants, 10 from plant collected in two squares, six from Anacardium occidentale L. - (Cashew). CSF was examined by direct microscopy by staining with Indian ink and cultured in Sabouraud agar with chloramphenicol at 25 °C and 37 ºC. In the period from 2005 to 2007 were obtained from 141 environmental isolates and35 clinical isolates of Cryptococcus neoformans species complex. Ecological and epidemiological inferences were determined. In addition characteristics of isolates were carried out by tests for presumptive phenotypic identification, through Niger medium and species into the CGB test, sensitivity to the antifungal, enzymatic characterization of biotypes of interest and/or marker lines, determining sexual mating type using PCR and PCR Fingerprintig with primer M13. Among the strains of Cryptococcus spp. isolated from CSF of patients with infectious meningitis in the state of Sergipe, in the period from 2001 to 2009, prevaled the species C. neoformans. Among the environmental strains, only one sample of 140, was identified as C. gattii. All samples from thefeces of birds were positive in the isolation of yeast. Clinical isolates ofCryptococcus neoformans and C. gattii originating from patients withcryptococcal meningitis in the state of Sergipe exhibited higher values ofminimum inhibitory concentration (MIC) to the antifungal agents tested, which symbolizes high rate of resistance, especially to amphotericin B and fluconazole. In relation to sexual mating type was evident that most of the clinics were MAT strains, confirming the character of virulence of this yeast. However, among the environmental isolates the MATa was more prevalent, showing the character of these less virulent strains. Cryptococcus neoformans was the only species amplified by PCR fingerprinting. There was a presence among the local samples of three different genetic profiles belonging to the same species.