Ana Roberta Fusco da Costa

Characterization of 17 strains belonging to the Mycobacterium simiae complex and description of Mycobacterium paraense sp. nov.

Fourteen mycobacterial strains isolated from pulmonary samples of independent patients in the state of Pará (Brazil), and three strains isolated in Italy, were characterized using a polyphasic approach. Thorough genetic investigation, including whole-genome sequencing, demonstrated that the strains belong to the M. simiae complex, being most closely related to Mycobacterium interjectum. For 14 of the strains, evidence emerged supporting their inclusion in a previously unreported species of the genus Mycobacterium, for which the name Mycobacterium paraense sp. nov. is proposed (type strain, IEC26(T) = DSM 46749(T) = CCUG 66121(T)). The novel species is characterized by slow growth, unpigmented or pale yellow scotochromogenic colonies, and a HPLC mycolic acid profile different from other known mycobacteria. In different genetic regions, high sequence microheterogeneity was detected.

Occurrence of Nontuberculous Mycobacterial Pulmonary Infection in an Endemic Area of Tuberculosis

The majority of investigations of the epidemiology of nontuberculous mycobacteria (NTM) have focused on highly developed nations with a low prevalence of tuberculosis. In contrast, the Para state of north Brazil represents an area of high tuberculosis prevalence and increasing NTM incidence. Toward the goal of understanding the dynamics of infection by all Mycobacterium species, we report patient characteristics and the identification of NTM strains isolated from sputum samples from patients that were residents of Para, a state in the Amazon region, Northern of Brazil, over the period January 2010 through December 2011 (2 years). The 29 NTM patients comprised 13.5% of positive mycobacterial cultures over the 2-year period. A major risk factor for NTM pulmonary disease was previous tuberculosis (76%). Further, the average age of NTM patients (52 years) was significantly higher than that of tuberculosis patients (39 years) and more were female (72.4% vs. 37.4%). Unlike other Brazilian states, NTM pulmonary patients in Para were infected with a different spectrum of mycobacteria; primarily the rapidly growing Mycobacterium massiliense and Mycobacterium simiae complex.

A fatal case of pulmonary infection by Mycobacterium colombiense in Para State, Amazon Region, Brazil

Mycobacterium avium complex (MAC) is a heterogeneous group of species found in several environmental sources and that exhibit variable degrees of pathogenicity. Among the MAC members, Mycobacterium colombiense has been related to pulmonary disease and disseminated infection in HIV-infected patients in Colombia. Lymphadenopathy cases have also been reported. We have described a fatal case of M. colombiense pulmonary disease in a Brazilian patient without evidence of HIV infection or other known causes of immunosuppression.

Análise da distribuição espaço-temporal da leptospirose humana em Belém, Estado do Pará, Brasil

INTRODUÇÃO: A leptospirose é um problema de saúde pública de interesse mundial, que tem elevados impactos econômicos e sociais. A doença está relacionada a áreas com precárias condições sanitárias, atingindo principalmente as periferias urbanas. No presente estudo descreveu-se a frequência de casos de leptospirose, indicando áreas de risco para sua ocorrência no Município de Belém, Estado do Pará, Brasil. METODOLOGIA: Foi realizado um estudo epidemiológico da leptospirose humana em Belém no período de 2006 a 2011, utilizando como base de dados os registros de casos confirmados constantes no Sistema de Informação de Agravos de Notificação (SINAN) da Secretaria de Estado de Saúde Pública do Estado do Pará. RESULTADOS: Cerca de 40% dos casos estavam concentrados em três bairros, todos com infraestrutura de saneamento básico deficiente. Observou-se uma tendência de redução da incidência na série histórica, apesar das taxas de letalidade ainda se manterem elevadas. A faixa etária de 15 a 39 anos concentrou 53,7% dos registros, com maior letalidade para indivíduos acima de 40 anos (19,8%, p = 0,0004). Em 48,1% deles a principal situação de risco associada ao adoecimento foi o contato prévio com água ou lama, e 45,1% tiveram o ambiente domiciliar como provável local de infecção. O bairro do Guamá apresentou a maior densidade de casos, sendo o extremo norte continental do Município sugerido como área de risco potencial para o estabelecimento da leptospirose. CONCLUSÃO: O presente estudo permitiu o delineamento de áreas de maior risco para ocorrência de casos, determinação de preditores de severidade, assim como as perdas sociais resultantes dos óbitos.

Genotipagem por spoligotyping de Mycobacterium tuberculosis obtidos de lâminas de Ziehl-Neelsen em Belém, Estado do Pará, Brasil

Este trabalho foi realizado com apoio financeiro do Instituto Evandro Chagas/ SVS/ MS, Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior e Programa de Iniciacao Cientifica/ IEC/ CNPq.

Pulmonary Nontuberculous Mycobacterial Infections in the State of Para, an Endemic Region for Tuberculosis in North of Brazil

Traditionally, Mycobacterium species are divided in those belonging to the Mycobacterium tuberculosis complex (MTBC), M. leprae and the nontuberculous or atypical mycobacteria (NTM), the latter consisting of either rapid (RGM) or slow growing (SGM) species, forming colonies, respectively, within seven days of culture or requiring longer incubation time (Runyon, 1959). Among the RGM are the M. chelonae complex, including M. chelonae, M. abscessus, M. mucogenicum, M. salmoniphilum, M. bolletii and M. massiliense (Brown-Elliot & Wallace, 2002; Whipps et al., 2007) and the M. fortuitum complex, including M. fortuitum, M. peregrinum, M. septicum, M. mageritense, M. houstonense and M. boenikei (Adekambi & Drancourt, 2004), containing the NTM commonly encountered in human specimens. Clinically important SGM are the M. avium complex (MAC), which include M. avium, M. intracellulare, M. colombiense and M. chimaera (Tortoli, 2003; Tortoli et al., 2004).

Annual frequency and distribution of tuberculosis resistance in the public health laboratory network of Pará State, Brazil

This study was supported by the Instituto Evandro Chagas, Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) and the Programa de Pos-graduacao em Biologia Parasitaria/Universidade Estadual do Para.

Identificação genotípica de membros do complexo Mycobacterium avium isolados de infecções pulmonares no Estado do Pará, Brasil

INTRODUCTION: The Mycobacterium avium complex (MAC) is a group of slow-growing mycobacteria naturally found in the environment capable of causing infections in a wide variety of living species, including birds, swines and humans. These infections can be asymptomatic, clinically significant and, in some cases, fatal. There is a demand for techniques that are capable of conclusively identifying closely related bacteria. Molecular biological techniques are promising tools for a more precise identification. MATERIAL AND METHODS: In this study, we evaluated the ability of 16S rRNA, hsp65 and rpoB molecular markers to distinguish between members of the MAC isolated in the Mycobacteria Laboratory at the Instituto Evandro Chagas. RESULTS: MAC samples collected from 15 patients were previously evaluated using an hsp65 gene restriction fragment length polymorphism analytical method (RFLP-hsp65), and they showed three different profiles: (I) BstEII: 235/115/100, HaeIII: 145/130/60; (II) BstEII: 235/210, HaeIII: 130/105; and (III) BstEII: 235/210, HaeIII: 145/130. We constructed phylogenetic trees using 16S rRNA analysis in which the samples were distributed into two groups, similarly to those found in the hsp65 analysis. However, the results from the rpoB analysis disagreed with those of the other trees due to changes in topology. CONCLUSION: The findings from this study warrant that various evolutionary forces may be acting on the rpoB gene. Thus, it is necessary to be cautious when using this target for taxonomic analyses. Additionally, we recommend that multiple markers (including16S rRNA) be evaluated when identifying mycobacteria.INTRODUCTION: The Mycobacterium avium complex (MAC) is a group of slow-growing mycobacteria naturally found in the environment capable of causing infections in a wide variety of living species, including birds, swines and humans. These infections can be asymptomatic, clinically significant and, in some cases, fatal. There is a demand for techniques that are capable of conclusively identifying closely related bacteria. Molecular biological techniques are promising tools for a more precise identification. MATERIAL AND METHODS: In this study, we evaluated the ability of 16S rRNA, hsp65 and rpoB molecular markers to distinguish between members of the MAC isolated in the Mycobacteria Laboratory at the Instituto Evandro Chagas. RESULTS: MAC samples collected from 15 patients were previously evaluated using an hsp65 gene restriction fragment length polymorphism analytical method (RFLP-hsp65), and they showed three different profiles: (I) BstEII: 235/115/100, HaeIII: 145/130/60; (II) BstEII: 235/210, HaeIII: 130/105; and (III) BstEII: 235/210, HaeIII: 145/130. We constructed phylogenetic trees using 16S rRNA analysis in which the samples were distributed into two groups, similarly to those found in the hsp65 analysis. However, the results from the rpoB analysis disagreed with those of the other trees due to changes in topology. CONCLUSION: The findings from this study warrant that various evolutionary forces may be acting on the rpoB gene. Thus, it is necessary to be cautious when using this target for taxonomic analyses. Additionally, we recommend that multiple markers (including16S rRNA) be evaluated when identifying mycobacteria.

Desenvolvimento de PCR multiplex para detecção e diferenciação de categorias de Escherichia coli diarreiogênicas

INTRODUCTION: Diarrheagenic Escherichia coli (DEC) are considered an important cause of diarrhea in developing countries. The correct identification of these microorganisms depends on their differentiation from non-pathogenic members of the intestinal microbiota. DEC can be classified into one of six categories according to their mechanism of pathogenicity. MATERIALS AND METHODS: Two multiplex PCR systems used to detect enteropathogenic (EPEC), enterotoxigenic (ETEC), enteroinvasive (EIEC) and Shiga Toxin-producing (STEC) E. coli were evaluated and described. RESULTS: Four categories of DEC were detected among isolates of E. coli obtained from individuals infected with HIV. EIEC and EPEC were among the most prevalent pathotypes. Furthermore, two STEC strains that were both stx1and eaepositive were identified. This is the first report of this kind of isolation in individuals infected with HIV in the State of Pará. Multiplex PCR proved to be an efficient, fast and reproducible technique for detection of DEC isolates. Both multiplex PCR systems described here produced results 100% similar to those obtained from individual PCR reactions. CONCLUSION: Given their simplicity, cost and efficiency, it is possible to use these protocols to expedite the molecular diagnosis of the distinct categories of DEC. In addition to facilitating the development of new research projects, these findings could support the epidemiological surveillance undertaken by public health agencies and institutes.INTRODUCTION: Diarrheagenic Escherichia coli (DEC) are considered an important cause of diarrhea in developing countries. The correct identification of these microorganisms depends on their differentiation from non-pathogenic members of the intestinal microbiota. DEC can be classified into one of six categories according to their mechanism of pathogenicity. MATERIALS AND METHODS: Two multiplex PCR systems used to detect enteropathogenic (EPEC), enterotoxigenic (ETEC), enteroinvasive (EIEC) and Shiga Toxin-producing (STEC) E. coli were evaluated and described. RESULTS: Four categories of DEC were detected among isolates of E. coli obtained from individuals infected with HIV. EIEC and EPEC were among the most prevalent pathotypes. Furthermore, two STEC strains that were both stx1and eaepositive were identified. This is the first report of this kind of isolation in individuals infected with HIV in the State of Pará. Multiplex PCR proved to be an efficient, fast and reproducible technique for detection of DEC isolates. Both multiplex PCR systems described here produced results 100% similar to those obtained from individual PCR reactions. CONCLUSION: Given their simplicity, cost and efficiency, it is possible to use these protocols to expedite the molecular diagnosis of the distinct categories of DEC. In addition to facilitating the development of new research projects, these findings could support the epidemiological surveillance undertaken by public health agencies and institutes.

A case of pulmonary infection due to Mycobacterium paraffinicum from the Amazon Region

M. paraffinicum, a slow-growing scotochromogenic mycobacterium that uses paraffinic hydrocarbons other than methane, i.e. inorganic carbon sources, was originally isolated from soil samples, but only in 2010 definitely achieved the species status. We have described here the first report of pulmonary disease due to M. paraffinicum in Amazon Region.