Abstract INTRODUCTION: We aimed to estimate the prevalence and transmission of drug-resistant tuberculosis in a high-burden Brazilian setting under directly observed therapy short-course strategy. METHODS: Isolates of culture-confirmed pulmonary tuberculosis patients from Guarulhos, Brazil, diagnosed in October 2007-2011 were subjected to drug susceptibility and IS6110-restriction fragment length polymorphism testing. RESULTS: The overall resistance prevalence was 11.5% and the multi-drug resistance rate was 4.2%. Twenty-six (43.3%) of 60 drug-resistant isolates were clustered. Epidemiological relationships were identified in 11 (42.3%) patients; 30.8% of the cases were transmitted in households. CONCLUSIONS: Drug-resistant tuberculosis was relatively low and transmitted in households and the community.

ABSTRACT Introduction: The Mycobacterium tuberculosis East African-Indian (EAI) spoligotyping family (belonging to lineage 1, Indo-Oceanic, defined by the region of deletion RD239) is distributed worldwide, but is more prevalent in Southeast Asia, India, and East Africa. Studies in Latin America have rarely identified EAI. In this study, we describe the occurrence of the EAI family in Brazil. Methods: EAI was identified in a systematic literature review of genetic diversity studies pertaining to M. tuberculosis in Brazil, as well as in a survey conducted in Salvador, Bahia, located in the northeastern region of this country. Results: The EAI6-BGD1 spoligotyping family and the EAI5 Spoligotype International Type (SIT) 1983 clade were the most frequently reported, with wide distribution of this particular clade described in Brazil. The distribution of other EAI spoligotyping patterns with broader worldwide distribution was restricted to the southeastern region of the country. Conclusions: EAI may be endemic at a low frequency in Brazil, with some clades indicating increased fitness with respect to this population.

Este é o primeiro estudo realizado na Bahia, Brasil, visando à descrição da estrutura da população genética circulante do Mycobacterium tuberculosis na cidade de Salvador. Um total de 56 casos confirmados de tuberculose pulmonar, identificados entre março e junho de 2008, foi analisado pelo método Restriction Fragment Lenght Polymorphism (IS6110-RFLP). A população de estudo foi caracterizada como a maioria do sexo masculino (71,43 %), idade acima de 30 anos (68,75%). Quarenta e um isolados (73,21%) com padrão único, enquanto 15 (26,75%) apresentaram padrões agrupáveis, formando seis clusters. A alta taxa de diversidade das cepas de M. tuberculosis observada é mais sugestiva de reativação endógena do que transmissão recente.

This study constitutes a first attempt to describe the genetic population structure of Mycobacterium tuberculosis circulating in Salvador, Bahia State, Brazil. A total of 56 confirmed cases of pulmonary tuberculosis, identified between March and June 2008, were analyzed using restriction fragment length polymorphism (IS6110-RFLP). The study population was characterized by a predominance of males (71.43%) over 30 years of age (68.75%). Forty-one isolates were found to belong to a single pattern (73.2%), while 15 (26.7%) were found in group patterns, forming six clusters. The higher level of diversity observed is much more suggestive of endogenous reactivation than recent transmission.

Este é o primeiro estudo realizado na Bahia, Brasil, visando à descrição da estrutura da população genética circulante do Mycobacterium tuberculosis na cidade de Salvador. Um total de 56 casos confirmados de tuberculose pulmonar, identificados entre março e junho de 2008, foi analisado pelo método Restriction Fragment Lenght Polymorphism (IS6110-RFLP). A população de estudo foi caracterizada como a maioria do sexo masculino (71,43 %), idade acima de 30 anos (68,75%). Quarenta e um isolados (73,21%) com padrão único, enquanto 15 (26,75%) apresentaram padrões agrupáveis, formando seis clusters. A alta taxa de diversidade das cepas de M. tuberculosis observada é mais sugestiva de reativação endógena do que transmissão recente.

This study constitutes a first attempt to describe the genetic population structure of Mycobacterium tuberculosis circulating in Salvador, Bahia State, Brazil. A total of 56 confirmed cases of pulmonary tuberculosis, identified between March and June 2008, were analyzed using restriction fragment length polymorphism (IS6110-RFLP). The study population was characterized by a predominance of males (71.43%) over 30 years of age (68.75%). Forty-one isolates were found to belong to a single pattern (73.2%), while 15 (26.7%) were found in group patterns, forming six clusters. The higher level of diversity observed is much more suggestive of endogenous reactivation than recent transmission.

A expressão das proteínas reguladas pelo ferro (IRPs), in vitro, tem sido obtida pela adição de quelantes de ferro ao meio de cultura, após o crescimento bacteriano, na presença de fonte de ferro orgânico. Neste estudo foram investigados aspectos da máxima expressão das IRPs de meningococo durante o crescimento normal, em condições de cultura definidas, utilizando-se o meio de Catlin e os caldos Mueller-Hinton e Tryptic Soja (TSB). Foram avaliadas as melhores condições para se obter vesículas de membrana externa (OMVs) contendo IRPs para uso em vacina de meningococo B. A expressão das IRPs variou entre as diferentes cepas com relação as diferentes concentrações de Etilenediamine Di-orto-Hidroxifenil-ácido acético (EDDA), de acordo com o meio de cultura e também entre os diferentes lotes de TSB. Para cada cepa, um específico padrão de ERPs foi expresso e altas concentrações de EDDA, adição de glicose ou o uso de diferentes meios de cultura não resultou em expressão diferencial das IRPs. Não foi possível cultivar meningococo em condições normais de crescimento utilizando-se Desferral. Foi investigado condições de crescimento para se obter bom rendimento de OMVs com expressão de IRPs em meio de Catlin deficiente em ferro, contendo EDDA e hemina. Cultura de 32 h a 30ºC, após incubação por 16 h a 37ºC, manteve bom crescimento, porém, lise da bactéria foi observada após 46 h a 30ºC. Aproximadamente 4 vezes mais OMVs foram recuperadas do sobrenadante de cultura após 24 h a 30ºC do que das células após 16 h a 37ºC. As IRPs foram bem expressas nas OMVs obtidas do sobrenadante de cultura após 24 h a 30ºC como também nas OMVs obtidas das células crescidas por 16 h a 37ºC.

The expression of iron regulated proteins (IRPs) in vitro has been obtained in the past by adding iron chelators to the culture after bacterial growth, in the presence of an organic iron source. We have investigated aspects concerning full expression of the meningococcal IRPs during normal growth, in defined conditions using Catlin medium, Mueller Hinton and Tryptic Soy Broth (TSB). The expression of IRPs varied between different strains with respect to Ethylenediamine Di-ortho-Hidroxy-phenyl-acetic acid (EDDA) concentrations, and according to culture medium, and also between different lots of TSB. For each strain, a specific set of IRPs were expressed and higher EDDA concentrations, or addition of glucose, or use of different culture media did not resulted in a differential expression of IRPs. We were not able to grow N. meningitidis under normal growth conditions using Desferal. We looked for a good yield of outer membrane vesicles (OMVs) expressing IRPs in iron-deficient Catlin medium containing EDDA and Hemin. Culture for 32 h at 30ºC after growing for 16 h at 37ºC supported good bacterial growth. Bacterial lysis was noted after additional 24 h at 30ºC. Approximately 4 times more OMVs was recoverable from a culture supernatant after 24 h at 30ºC than from the cells after 16 h at 37ºC. The IRP were as well expressed in OMVs from culture supernatant obtained after 24 h at 30ºC as from the cells after 16 h at 37ºC.