ABSTRACT A drug resistance survey involving Mycobacterium tuberculosis isolated from patients of a tertiary Hospital in the Rio de Janeiro city (RJ), Brazil, between the years 1996 and 1998 revealed a high frequency of isoniazid (HR) resistance. These isolates were revisited and genotyped. Patients came from different RJ neighborhoods and municipalities, and 70% were outpatients. Applying the 3’ and 5’ IS 6110 -RFLP and the Spoligotype genotyping methods, the clonal structure of this population was investigated obtaining a snapshot of past epidemiological events. The 3’ clusters were subsequently 5’ IS 6110 -RFLP typed. Spoligotyping was analyzed in the SITVIT2 database. Epidemiological relationships were investigated. The major lineage was T (54.4%), and SIT 53/T1 and SIT 535/T1 were the most frequent. The T1 sublineage comprises 12.8% of resistant strains and SIT 535 were assigned for 31.8% of them. Orphan patterns corresponded to 12% and 73.3% and belonged to the T lineage. One pattern was unlisted in the SITVIT2. The 5’ IS 6110 -RFLP did not confirm 3/12 of the 3’ IS 6110 -RFLP clusters. A combination of all methods decreased the number of clusters to three. Nosocomial transmission was associated with one cluster involving a hospital cupbearer. This event was suspected in a multidrug resistant-TB inpatient caregiver who harbored a mixed infection. The 3’ IS 6110 clusters were associated with HR (p=0.046). These genotypic retrospective data may reflect a fraction of more extensive recent transmission in different communities that may be corroborated by the concentration of HR patients, and may serve as a database for further evolutionary and characterization evaluation of circulating strains and together with epidemiological data favors a more effective transmission control.

The purpose of this study was to provide information about the genetic diversity and prevalent genotype of Mycobacterium tuberculosis in a low-endemic setting in northwestern state of Paraná in Southern Brazil. We employed spoligotyping and mycobacterial interspersed repetitive units-variable number tandem repeat (MIRU-VNTR) techniques to genotype M. tuberculosisisolates from patients with pulmonary tuberculosis (TB). The 93 isolates analyzed by spoligotyping were divided into 36 different patterns, 30 of which were described in the SITVIT database. Latin American and Mediterranean, Haarlem and T families were responsible for 26.9%, 17.2% and 11.8% of TB cases, respectively. From the 84 isolates analyzed by MIRU-VNTR, 58 shared a unique pattern and the remaining 26 belonged to nine clusters. The MIRU loci 40, 23, 10 and 16 were the most discriminatory. A combination of MIRU-VNTR and spoligotyping resulted in 85.7% discriminatory power (Hunter-Gaston index = 0.995). Thus, combining spoligotyping and MIRU-VNTR typing proved to be most useful for epidemiological study in this low-endemic setting in Southern Brazil. The current study demonstrated that there is significant diversity in circulating strains in the city of Maringá and the surrounding regions, with no single genotype of M. tuberculosispredominating.

Estudos epidemiológicos sobre giardíase por técnicas moleculares como a RAPD (Amplificação aleatória do DNA polimórfico) podem contribuir para o entendimento de fatores relacionados à transmissão de Giardia duodenalis. O objetivo desse trabalho foi estudar a epidemiologia de G. duodenalis em 101 crianças atendidas em uma creche em Presidente Bernardes, SP, Brasil. Após exames parasitológicos em amostras de fezes, 15 crianças apresentaram cistos de G. duodenalis. Seus respectivos pais, irmãos e animais domésticos, além dos funcionários da creche, foram submetidos a exames parasitológicos de fezes. Destes, sete mães e nove irmãos apresentaram G. duodenalis, enquanto os pais, funcionários da creche e animais de estimação (cães) não apresentaram o parasita. Além dos 15 casos com G. duodenalis, outras 23 crianças apresentaram outros enteroparasitas (Entamoeba coli, Endolimax nana, Enterobius vermicularis, Ascaris lumbricoides e Trichuris trichiura). As amostras contendo cistos de G. duodenalis das crianças e de seus parentes foram submetidas à análise molecular por RAPD após extração do DNA genômico e amplificação de um fragmento de uma região do rDNA 18S por PCR. Entre os 31 isolados de G. duodenalis (crianças e suas respectivas mães e irmãos), concluiu-se que a transmissão dos parasitas ocorreu entre as crianças, provavelmente durante seu convívio na creche, mas não foi decorrente do convívio familiar ou animais de estimação.

Epidemiological studies on giardiasis by using molecular techniques such as RAPD (Randomly Amplified Polymorphic DNA) may give information on factors related to the transmission of Giardia duodenalis. The aim of this work was to assess the epidemiology of G. duodenalis in 101 children attended at a daycare center in Presidente Bernardes, SP, Brazil. After parasitological examinations in feces samples, 15 children presented cysts of G. duodenalis. Their respective parents, brothers and pets, besides the daycare center workers, also had their feces submitted to parasitological analysis. Seven mothers and nine brothers also presented G. duodenalis cysts, while fathers, daycare workers and pets (dogs) did not presented the parasite. Besides the 15 cases with G. duodenalis, other 23 children presented other enteroparasites (Entamoeba coli, Endolimax nana, Enterobius vermicularis, Ascaris lumbricoides and Trichuris trichiura). Samples of G. duodenalis cysts from children and their relatives were submitted to molecular typing by RAPD after genomic DNA extraction and amplification of a fragment of the 18S rDNA region by PCR. After examining 31 isolates of G. duodenalis (children and their respective mothers and brothers), it was concluded that the parasite transmission occurred in children, probably during daily cohabitation at the daycare center, but not at home among their relatives or pets.

OBJETIVO: Investigar a prevalência de infecção, doença e eventual surto institucional de tuberculose em um hospital psiquiátrico empregando teste tuberculínico, pesquisa de micobactéria em material da árvore respiratória e técnica de rastreamento molecular baseada na seqüência IS6110. MÉTODOS: Durante o período de fevereiro a agosto de 2002, foi realizado teste tuberculínico entre os 74 internos e os 31 funcionários de um hospital psiquiátrico na Cidade de Rio Verde (GO), pesquisa de Mycobacterium tuberculosis em material da árvore respiratória dos pacientes e estudo do perfil genético pelo método restriction fragment length polymorphism - IS6110. RESULTADOS: Micobactérias foram isoladas de 5 pacientes analisados (6,8 %), sendo em quatro M. tuberculosis e em um M. chelonae. Os isolados de M. tuberculosis foram sensíveis a isoniazida e rifampicina e, através da técnica restriction fragment length polymorphism - IS6110, apresentaram perfis genéticos únicos, totalmente distintos uns dos outros, indicando que os casos de tuberculose encontrados eram decorrentes de reativação endógena, não sendo possível caracterizá-los como um surto institucional. Através da realização do teste tuberculínico em dois passos nos pacientes, verificaram-se taxas de infecção de 23% e 31%. Entre os funcionários a taxa foi de 42%, não tendo sido realizada entre estes a repetição do teste. CONCLUSÃO: Os resultados indicaram uma incidência considerável de tuberculose infecção entre os pacientes e funcionários, sendo também elevada a ocorrência de tuberculose doença entre os pacientes.

OBJECTIVE: To investigate the prevalence of infection, disease and eventual institutional outbreak of tuberculosis in a psychiatric hospital using the PPD test, as well as testing for mycobacteria in material collected from the respiratory tree and using molecular tracking technique based on insertion sequence 6110 (IS6110). METHODS: Between February and August of 2002, PPD tests were given to 74 inpatients and 31 staff members at a psychiatric hospital in the city of Rio Verde, located in the state of Goiás, Brazil. In addition, respiratory tree material collected from the inpatients was submitted to testing for Mycobacterium tuberculosis. RESULTS: Among the patients analyzed, mycobacteria were isolated from five (6.8%): four identified as M. tuberculosis and one as M. chelonae. The M. tuberculosis isolates were sensitive to isoniazid and rifampicin, and, when submitted to the restriction fragment length polymorphism/IS6110 technique, presented unique genetic profiles, totally distinct from one another, suggesting that all of the tuberculosis cases were due to endogenous reactivation. It was not possible to characterize this group of cases as an institutional outbreak. Performing the two-step tuberculin test in the patients, the infection rates were 23% and 31%, compared with 42% among staff members, who were submitted to the one-step test. CONCLUSION: The results indicate a high incidence of tuberculosis infection among inpatients and hospital staff, as well as a high occurrence of the disease among inpatients.