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Serratia marcescens is microorganism often associated with nosocomial outbreaks and opportunistic infections including, bloodstream infections, urinary tract infections, central nervous system infection and pneumonias...

Objective: To describe the clinical manifestations of patients with pulmonary infection caused by mycobacteria of the Mycobacterium abscessus complex (MABSC), and to compare these manifestations with those of patients infected with other nontuberculous mycobacteria (NTM). Methods: This was a retrospective cohort study involving 43 patients divided into two groups: the MABSC group, consisting of patients with pulmonary infection caused by MABSC (n = 17); and the NTM group, consisting of patients with pulmonary infection caused by NTM other than MABSC (n = 26). Patients were previously treated with a regimen of rifampin, isoniazid, pyrazinamide, and ethambutol before the diagnosis of NTM was confirmed by two culture-positive sputum samples. The nucleotide sequences of the hsp65, 16S rRNA, and/or rpoB genes were analyzed to identify the mycobacteria. Data were collected on demographic, clinical, and radiological characteristics, as well as on treatment responses and outcomes. Results: Loss of appetite was the only clinical manifestation that was significantly more common in the MABSC group than in the NTM group (p = 0.0306). The chance of having to use a second treatment regimen was almost 12 times higher in the MABSC group than in the NTM group. Treatment success was significantly higher in the NTM group than in the MABSC group (83.2% vs. 17.6%; p < 0.0001). The chance of recurrence was approximately 37 times higher in the MABSC group than in the NTM group. Conclusions: In the study sample, treatment response of pulmonary disease caused by MABSC was less favorable than that of pulmonary disease caused by other NTM. RESUMO Objetivo: Descrever as manifestações clínicas de pacientes com infecção pulmonar por micobactérias do complexo Mycobacterium abscessus (CMA) e compará-las com as daqueles infectados com outras micobactérias não tuberculosas (MNT). Métodos: Estudo de coorte retrospectivo envolvendo 43 pacientes divididos em dois grupos: grupo CMA, com pacientes com infecção pulmonar por CMA (n = 17); e grupo MNT, com pacientes com infecção pulmonar por MNT que não CMA (n = 26). Os pacientes foram previamente tratados com o esquema rifampicina, isoniazida, pirazinamida e etambutol antes de o diagnóstico de MNT ser confirmado com culturas positivas em duas amostras de escarro diferentes. As sequências nucleotídicas dos genes hsp65, RNAr 16S e/ou rpoB foram analisadas para a identificação das micobactérias. Foram coletadas características demográficas, clínicas e radiológicas, assim como respostas terapêuticas e desfechos. Resultados: A única manifestação clínica significativamente mais frequente no grupo CMA que no grupo MNT foi hiporexia (p = 0,0306). A chance de haver a necessidade de utilização de um segundo esquema terapêutico foi quase 12 vezes maior no grupo CMA que no grupo MNT. O sucesso terapêutico foi significativamente maior no grupo MNT que no grupo CMA (83,2% vs. 17,6%; p < 0,0001). A chance de recidiva no grupo CMA foi aproximadamente 37 vezes maior que no grupo MNT. Conclusões: Na amostra estudada, a resposta terapêutica da doença pulmonar causada por CMA evoluiu de forma menos favorável do que naquela causada pelas demais MNT.

This study was funded by the Evandro Chagas Institute. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Micologia. Ananindeua, PA, Brasil / Federal University of Par?. Biological Sciences Center. Postgraduate Program in Biology of Infectious and Parasitic Agents. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Micologia. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Micologia. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Micologia. Ananindeua, PA, Brasil / Federal University of Par?. Biological Sciences Center. Postgraduate Program in Biology of Infectious and Parasitic Agents. Bel?m, PA, Brazil. Recently, Trichosporon taxonomy has been reevaluated and new genera of the Trichosporonaceae family have been described. Here, 26 clinical isolates were submitted for identification via sequencing of the intergenic space 1 (IGS1) region, genotyping, and investigation of virulence factors. Antifungal susceptibility was determined using the CLSI broth microdilution method for fluconazole (FLC), itraconazole (ITC), and amphotericin B (AMB). Of these, 24 isolates were identified, including 12 T. asahii, 4 T. inkin, 3 T. faecale, 1 T. coremiiforme, 1 T. japonicum, 2 Cutaneotrichosporon dermatis (formerly T. dermatis), and 1 Apiotrichum mycotoxinivorans (formerly T. mycotoxinivorans). Species-level identification of 2 isolates was not successful; they were described as Trichosporon sp. We observed optimal colonial development at 35-40 ?C. Lipase was the major extracellular enzyme produced (100%); caseinase was not produced (0%). Biofilms were produced by all isolates (classified as low). High AMB minimum inhibitory concentration (MIC) was observed, with all strains resistant. Fluconazole was the most active drug among the antifungals tested. However, high MICs for FLC were observed in C. dermatis and A. mycotoxinivorans species, which also showed resistance to ITC and AMB. This study, conducted in the Northern region of Brazil, identified 5 Trichosporon species along with C. dermatis and A. mycotoxinivorans and demonstrated their pathogenic potential through their ability to produce important virulence factors. This may contribute to our understanding of the epidemiology and factors related to the pathogeneses of species in the Trichosporonaceae family.

Funda??o de Amparo ? Pesquisa no Par?/Universidade do Estado do Par? (FAPESPA/UEPA) [003/2014], Instituto Evandro Chagas/Minist?rio da Sa?de/Secretaria de Vigil?ncia em Sa?de (IEC/MS/SVS), Programa Institucional de Bolsas de Inicia??o Cient?fica/Instituto Evandro Chagas - Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (PIBIC/IEC-CNPq) and Funda??o para o desenvolvimento cient?fico e tecnol?gico em sa?de [PRES-012-FIO-16]. MLC is supported by the Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior ? Brazil (CAPES) ? Finance Code 001, [194 - 88881.187587/2018-01, 2018]. Universidade do Estado do Par?. Centro de Ci?ncias Biol?gicas e da Sa?de. Programa de P?s-Gradua??o Biologia Parasit?ria na Amaz?nia. Bel?m, PA, Brazil. Universidade do Estado do Par?. Centro de Ci?ncias Biol?gicas e da Sa?de. Programa de P?s-Gradua??o Biologia Parasit?ria na Amaz?nia. Bel?m, PA, Brazil / Universidade de Lisboa. Faculdade de Farm?cia. Lisboa, Portugal. Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Paulo de G?es. Rio de Janeiro, RJ, Brazil / Funda??o Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laborat?rio de Bacteriologia e Bioensaios. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade do Estado do Par?. Centro de Ci?ncias Biol?gicas e da Sa?de. Programa de P?s-Gradua??o Biologia Parasit?ria na Amaz?nia. Bel?m, PA, Brazil. Universidade do Estado do Par?. Centro de Ci?ncias Biol?gicas e da Sa?de. Bel?m, PA, Brazil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Laborat?rio de Biologia Molecular Aplicada ? Microbact?ria. Rio de Janeiro, RJ, Brazil. Funda??o Oswaldo Cruz. Instituto Oswaldo Cruz. Laborat?rio de Biologia Molecular Aplicada ? Microbact?ria. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Geoprocessamento. Ananindeua, PA, Brasil. Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Paulo de G?es. Rio de Janeiro, RJ, Brazil Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Paulo de G?es. Rio de Janeiro, RJ, Brazil Universidade Federal Rural da Amaz?nia. Instituto da Sa?de e Produ??o Animal. Bel?m, PA, Brazil. Universidade Federal Rural da Amaz?nia. Instituto da Sa?de e Produ??o Animal. Bel?m, PA, Brazil. Universidade Federal do Rio de Janeiro. Instituto de Estudos em Sa?de Coletiva. Programa de P?s-Gradua??o em Sa?de Coletiva. Rio de Janeiro, RJ, Brazil. Universidade Federal Rural da Amaz?nia. Programa P?s-gradua??o em Sa?de e Produ??o Animal na Amaz?nia. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade do Estado do Par?. Centro de Ci?ncias Biol?gicas e da Sa?de. Programa de P?s-Gradua??o Biologia Parasit?ria na Amaz?nia. Bel?m, PA, Brazil / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Mycobacterium bovis is the main causative agent of bovine tuberculosis (bTB) being among the animal-adapted Mycobacterium tuberculosis complex. Herds can also be infected with non-tuberculous mycobacteria (NTM) causing a negative effect on the economy and on animal and human health through zoonotic infections. Molecular tools are required for mycobacteria identification; thus, it is laborious to determine the epidemiological information of mycobacteria among herds. We aimed to describe the mycobacterial pathogens associated with cases of suspected bTB lesions in cattle/buffaloes slaughtered for consumption and to investigate bTB transmission. We evaluated 74 lesion samples from 48 animals (27 bovine/21 buffaloes) from 16 mapped farms. Positives samples from nested-PCR were cultured in Lowenstein-Jensen (LJ), 2% pyruvate (LJ?+?P), and 2% glycerol (LJ?+?G) media, followed by Ziehl-Neelsen (ZN) staining technique and partial gene sequencing (hsp65, rpoB, and 16S-rRNA). Spoligotyping and 24-MIRU-VNTR were performed. The LJ?+?P increased the chance of obtaining bacilli. The respiratory tract and the oral cavity were the most important infection route. In addition, the calcified part of the lesions suggested chronic bTB. Spoligotypes of M. bovis (SIT986/SB0885) differed from others found in South America, and the MIRU-VNTR 24 loci suggested that bTB was associated to highly related strains. The NTM species found are of clinical importance in humans.

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Federal Rural da Amaz?nia. Laborat?rio de Investiga??o e Diagn?stico de Enfermidades Animal. Bel?m, PA, Brasil. Universidade Federal Rural da Amaz?nia. Laborat?rio de Investiga??o e Diagn?stico de Enfermidades Animal. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. O presente trabalho relata um surto de leptospirose ocorrido no plantel de ovinos criados com finalidade de fornecimento de sangue aos laborat?rios para produ??o de meios de cultivos e de testes de fixa??o de complemento. Foram realizadas an?lises sorol?gica, molecular e histopatol?gica. A an?lise sorol?gica foi realizada em 12 amostras utilizando o teste de soroaglutina??o microsc?pica com ponto de corte 100, para os sorovares Leptospira Bataviae, L. Javanica, L. Panama, L. Hebdomadis, L. Castellonis, L. Icterohaemorrhagiae, L. Pyrogenes, L. Cynopteri, L. Serjoe, L. Australis, L. Wolffi, L. Copenhageni, L. Autumnalis, L. Pomona e L. Tarassovi. O exame anatomopatol?gico foi realizado somente em um dos animais, o qual foi encontrado morto e as an?lises moleculares foram realizadas no sangue e na urina utilizando a t?cnica do qPCR. Foram reagentes ao diagn?stico sorol?gico 50% dos animais, sendo identificados os sorovares L. Copenhageni (1/100 e 1/200), L. Castellonis (1/200 e 1/200), L. Icterohaemorrhagiae (1/200) e L. Cynopteri (1/200). O animal necropsiado apresentou hemorragia, icter?cia e hemoglobin?ria. A an?lise molecular demonstrou tr?s animais positivos em amostras de sangue e dois animais positivos em amostras de urina, n?o havendo positividade concomitante para um mesmo animal. A identifica??o dos sorovares Icterohaemorrhagiae e Copenhageni sugere a necessidade de estudos, visando o isolamento do agente na regi?o e a caracteriza??o de sua patogenicidade, visto que no Brasil estes s?o os principais agentes etiol?gicos dos casos de leptospirose em humanos. The current paper reports an outbreak of leptospirosis in sheep production created with the purpose of blood supply to laboratories for production of growth medium and complement fixation test. Serological, molecular and histopathological analyses were performed. Serologic analysis was performed on 12 samples using the microscopic agglutination test with cutoff point of 100 for the serovars: Leptospira Bataviae, L. Javanica, L. Panama, L. Hebdomadis, L. Castellonis, L. Icterohaemorrhagiae, L. Pyrogenes, L. Cynopteri, L. Serjoe, L. Australis, L. Wolffi, L. Copenhageni, L. Autumnalis, L. Pomona e L. Tarassovi. Anatomopathological examination was performed on only one animal, which was found dead and molecular analyzes were performed in its blood and urine using the qPCR technique. 50% of those animals were reagents to the serological diagnosis, being identified the serovars: L. Copenhageni (1/100 e 1/200), L. Castellonis (1/200 e 1/200), L. Icterohaemorrhagiae (1/200) e L. Cynopteri (1/200). The necropsied animal presented bleeding, jaundice and hemoglobinuria. Molecular analysis showed three positive animals in blood samples and two positive animals in urine samples, with no concomitant positivity for each animal. The identification of Icterohaemorrhagiae and Copenhageni serovars suggests the need for studies aiming the agent isolation in the Brazilian Amazon Region and characterization of its pathogenicity since in Brazil these are the main etiologic agents of leptospirosis cases in humans.

Federal University of Para. Belem, PA, Brazil. University Center of Para. Belem, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. 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.

Federal University of Par?. Belem, PA, Brazil. Federal University of Par?. Belem, PA, Brazil. Federal University of Par?. Belem, PA, Brazil. Federal University of Par?. Belem, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. 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.

Este trabalho foi realizado com apoio financeiro do Instituto Evandro Chagas/ SVS/ MS, Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior e Programa de Inicia??o Cient?fica/ IEC/ CNPq. Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil. Universidade do Estado do Par?. Programa de P?s-gradua??o em Biologia Parasit?ria da Amaz?nia. Bel?m, PA, Brasil. Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Geoprocessamento. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Geoprocessamento. Ananindeua, PA, Brasil. Funda??o Oswaldo Cruz. Laborat?rio de Biologia Molecular Aplicada a Micobact?rias. Rio de Janeiro, RJ, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. O spoligotyping ? uma t?cnica molecular baseada na amplifica??o de uma regi?o de repeti??es diretas polim?rficas, a qual permite detectar e diferenciar Mycobacterium tuberculosis (MT). Neste estudo, foram genotipados MT obtidos de material fixado em l?minas coradas pela t?cnica de Ziehl-Neelsen (ZN), confeccionadas em laborat?rios p?blicos de Bel?m. A identifica??o molecular de MT por spoligotyping em amostras de 102 indiv?duos apresentou elevada sensibilidade, com 90 padr?es de hibridiza??o completos e concordantes entre si. A classifica??o dos gen?tipos foi realizada comparando os resultados obtidos com aqueles dispon?veis no banco de dados internacional SITVITWEB. Foram observados 45 gen?tipos distintos, dos quais 35 previamente relatados e 11 ainda n?o relatados. Sessenta e um gen?tipos apresentaram-se compartilhados por duas a 15 amostras e 29 gen?tipos eram ?nicos. As fam?lias LAM, T, H e EAI foram as mais frequentes. Os bairros com maior concentra??o de casos foram Guam?, Jurunas e Terra Firme. A genotipagem por spoligotyping torna-se uma importante ferramenta no monitoramento de isolados em diferentes contextos epidemiol?gicos. A possibilidade de caracterizar gen?tica e demograficamente estes microorganismos contribui para o melhor entendimento de como a doen?a ? distribu?da nesta popula??o, no esclarecimento das transmiss?es e das contamina??es cruzadas e na implementa??o de a??es para o controle da tuberculose. Spoligotyping is a molecular method based on the amplification of a polymorphic direct repeat locus, which allows for the detection and identification of Mycobacterium tuberculosis. Samples of M. tuberculosis obtained from material fixed in Ziehl-Neelsen-stained slides from public laboratories in Bel?m, Par? State, Brazil, were genotyped in this study. The molecular characterization of M. tuberculosis through spoligotyping of samples from 102 patients showed high sensitivity, with 90 complete and matching hybridization patterns. The genotypes were classified based on a comparison of the results obtained with data available in the SITVITWEB international database. Of the 45 different genotypes observed, 35 had already been reported and 11 had never been reported. Sixty-one genotypes were observed in 2 to 15 samples, and 29 genotypes did not match. LAM, T, H and EAI were the most common families. The districts with the highest concentration of cases were Guam?, Jurunas and Terra Firme. Spoligotyping is considered an important tool to follow up isolates in distinct epidemiologic contexts. Genetic and demographic characterization of these microorganisms contributes to a better understanding of the distribution of tuberculosis in this population, its transmission process, the occurrence of cross contaminations, and the implementation of controlling measures of the disease.

This study was supported by the Instituto Evandro Chagas, Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) and the Programa de P?s-gradua??o em Biologia Parasit?ria/Universidade Estadual do Par?. Universidade do Estado do Par?. Programa de P?s-gradua??o em Biologia Parasit?ria da Amaz?nia. Bel?m, PA, Brasil. Universidade do Estado do Par?. Programa de P?s-gradua??o em Biologia Parasit?ria da Amaz?nia. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Geoprocessamento. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. This study described the frequency and variety of multidrug-resistant tuberculosis (MDR-TB) cases in Par? State, Brazil. In order to record the epidemiology and spacial distribution of the disease, antimicrobial susceptibility testing (AST) and the proportion method were used. The AST was performed on 848 samples at the Instituto Evandro Chagas and a central public health laboratory in that State. Of all patients enrolled in the study, 358 (42.2%) were resistant to at least one antituberculosis (anti-TB) drug. The percentage of primary, acquired resistance and combined MDR-TB was 30.4%, 69.3% and 42.2% respectively. MDR-TB was detected in 223 (26.3%) samples. Primary MDR-TB was found in 14% of previously treated patients, while 48% exhibited acquired resistance and 62% exhibited combined resistance. Of all age groups, from 25 to 36 years old (mean age 38.7 ? 15) showed the highest proportion of resistant cases (26.3%). Seven cities in the State presented 59.6% of the cases documented in the study. These findings reflect the poor quality of the healthcare for patients in these cities. We further suggest that clinicians need to observe their patients more directly during treatment and test them more often for anti-TB drug sensitivity. The number of TB cases in Par? did not vary significantly during the period studied, but researchers did notice a slight increase in the proportion of drug-resistant cases related to the total number of cases reported in Par?. This change in the resistance rates reflects the need to improve the quality of health services for TB care. A concentration of TB cases was observed in some municipalities and in the neighborhoods of the Bel?m City. Este estudo descreveu a frequ?ncia e variedade de casos de tuberculose multirresistente (TBMR) no Estado do Par?, Brasil. A fim de registrar a epidemiologia e a distribui??o espacial da doen?a, foram utilizados testes de suscetibilidade antimicrobiana (TSA) e do m?todo proporcional (MP). O TSA foi realizado em 848 amostras no Instituto Evandro Chagas e em um laborat?rio central de sa?de p?blica naquele Estado. De todos os pacientes inclu?dos no estudo, 358 (42,2%) eram resistentes a pelo menos um medicamento antituberculose (anti-TB). A porcentagem das resist?ncias prim?ria, adquirida e TBMR foi de 30,4%, 69,3% e 42,2%, respectivamente. A TBMR foi detectada em 223 (26,3%) amostras. A TBMR prim?ria foi encontrada em 14% dos pacientes tratados anteriormente, enquanto que 48% apresentaram a resist?ncia adquirida e 62% exibiram resist?ncia combinada. De todos os grupos et?rios, o de 25-36 anos (m?dia de idade de 38,7 ? 15) apresentou a maior propor??o de casos resistentes (26%). Sete cidades do Estado apresentaram 59,6% dos casos documentados no estudo. Estes resultados refletem a baixa qualidade dos cuidados de sa?de com os doentes nestas cidades. Observa-se ainda, a necessidade de maiores cuidados dos m?dicos para com os pacientes durante o tratamento e realiza??o, com maior frequ?ncia, dos testes de sensibilidade aos medicamentos anti-TB. O n?mero de casos de TB no Par? n?o variou significativamente durante o per?odo estudado, por?m os pesquisadores notaram um ligeiro aumento na propor??o de casos resistentes aos medicamentos relacionados com o n?mero total de casos notificados no Estado. Esta mudan?a nas taxas de resist?ncia reflete a necessidade de melhorar a qualidade dos servi?os de sa?de para a aten??o ? TB. A concentra??o de casos de tuberculose foi observada em alguns munic?pios e nos bairros da Cidade de Bel?m.

This study utilized the hsp65 polymerase chain reaction restriction analysis (PRA) method in the identification of nontuberculous mycobacteria (NTMs) isolated in a Brazilian mycobacteria laboratory. NTM isolates from clinical specimens collected from 192 patients were characterized using the hsp65 PRA method and analyzed using both 16S rRNA and hsp65 gene sequencing. Only 30% of the NTM strains were correctly identified through PRA, though the suggested inclusion of an additional restriction enzyme could increase the resolution to roughly 90%. A total of 17 NTM strains were not identified to species level and may represent a new taxonomic entity classified as belonging to the Mycobacterium simiae complex. This study demonstrates the applicability of hsp65 PRA in the identification of several NTM strains in a reference laboratory, though the results suggest that some modifications to the original PRA method could increase its resolution substantially.

We isolated 44 strains of rapidly growing mycobacteria (RGM) from 19 patients with pulmonary infections assisted at the Instituto Evandro Chagas (Pará, Brazil) from 2004 to 2007. Identification at the species level was performed by PCR restriction fragment length polymorphism analysis (PRA) of a 441 bp hsp65 fragment and partial 16S rRNA, hsp65, and rpoB gene sequencing. Genotyping by PRA yielded 3 digestion patterns: one identical to Mycobacterium abscessus type I (group I); another to M. abscessus type II, Mycobacterium bolletii, and Mycobacterium massiliense (group II); and a third typical for Mycobacterium fortuitum type I (group III). When comparing analysis of the 3 genes, more discrimination was obtained by rpoB gene sequence, which allowed good distinction between group I, II, and III strains and subclassification of group II strains in SG IIa (M. bolletii) and SG IIb (M. massiliense). In this study, we show that the description of new RGM species requires the establishment of standardized procedures for RGM identification and the alert of the clinician about their involvement in pulmonary disease and the necessity of treatment for control and cure.

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, IEC26T = DSM 46749T = CCUG 66121T). 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.

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).

Secretaria de Estado de Sa?de P?blica do Par?. Divis?o de Vigil?ncia em Sa?de. Departamento de Controle de Doen?as Transmitidas por Vetores. Bel?m, PA, Brasil. Secretaria de Estado de Sa?de P?blica do Par?. Divis?o de Vigil?ncia em Sa?de. Departamento de Controle de Doen?as Transmitidas por Vetores. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Centro Universit?rio do Par?. Bel?m, PA, Brasil. Centro Universit?rio do Par?. Bel?m, PA, Brasil. Sistema de Prote??o da Amaz?nia. Divis?o de Prote??o Ambiental. Bel?m, PA, Brasil. Instituto de Estudos Superiores da Amaz?nia. Bel?m, PA, Brasil. Sistema de Prote??o da Amaz?nia. Coordena??o de Opera??es de Centro Regional de Bel?m. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. INTRODUCTION: Leptospirosis is a major health problem, which is responsible for severe financial and social impacts worldwide. Its occurrence is directly related to areas with precarious sanitary conditions, especially the ones located in the suburbs. This work assessed the frequency of cases of leptospirosis and pointed out its risk areas in the Municipality of Bel?m, Par? State, Brazil. METHODS: An epidemiologic study on human leptospirosis was carried out in Bel?m between 2006 and 2011, based on data about confirmed cases provided by the Information System on Diseases of Compulsory Declaration (Sistema de Informa??o de Agravos de Notifica??o ? SINAN) of the Par? State Secretary of Health. RESULTS: Approximately 40% of the confirmed cases were concentrated in three districts with poor sanitary infrastructure. Although the lethality rates remained high, a reduction trend in the incidence historical series was observed. The age group between 15 and 39 years accounted for 53.7% of the cases reported, and individuals older than 40 years showed the highest lethality rate (19.8%, p = 0.0004). Previous contact with contaminated water and mud was the main risk factor for 48.1% of the cases, and the domestic environment was the most probable locus of infection for 45.1%. Guam? district presented the highest density of cases, and the northernmost part of the Municipality was considered a potential risk area for the establishment of leptospirosis. CONCLUSION: This study outlined the high-risk areas for the occurrence of leptospirosis in the Municipality of Bel?m. It also determined the predictive factors of severity of the disease and the social losses due to its mortality. INTRODU??O: A leptospirose ? um problema de sa?de p?blica de interesse mundial, que tem elevados impactos econ?micos e sociais. A doen?a est? relacionada a ?reas com prec?rias condi??es sanit?rias, atingindo principalmente as periferias urbanas. No presente estudo descreveu-se a frequ?ncia de casos de leptospirose, indicando ?reas de risco para sua ocorr?ncia no Munic?pio de Bel?m, Estado do Par?, Brasil. METODOLOGIA: Foi realizado um estudo epidemiol?gico da leptospirose humana em Bel?m no per?odo de 2006 a 2011, utilizando como base de dados os registros de casos confirmados constantes no Sistema de Informa??o de Agravos de Notifica??o (SINAN) da Secretaria de Estado de Sa?de P?blica do Estado do Par?. RESULTADOS: Cerca de 40% dos casos estavam concentrados em tr?s bairros, todos com infraestrutura de saneamento b?sico deficiente. Observou-se uma tend?ncia de redu??o da incid?ncia na s?rie hist?rica, apesar das taxas de letalidade ainda se manterem elevadas. A faixa et?ria de 15 a 39 anos concentrou 53,7% dos registros, com maior letalidade para indiv?duos acima de 40 anos (19,8%, p = 0,0004). Em 48,1% deles a principal situa??o de risco associada ao adoecimento foi o contato pr?vio com ?gua ou lama, e 45,1% tiveram o ambiente domiciliar como prov?vel local de infec??o. O bairro do Guam? apresentou a maior densidade de casos, sendo o extremo norte continental do Munic?pio sugerido como ?rea de risco potencial para o estabelecimento da leptospirose. CONCLUS?O: O presente estudo permitiu o delineamento de ?reas de maior risco para ocorr?ncia de casos, determina??o de preditores de severidade, assim como as perdas sociais resultantes dos ?bitos.

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Federal do Par?. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Federal do Par?. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. INTRODU??O: O complexo Mycobacterium avium (MAC) compreende micobact?rias de crescimento lento, naturalmente encontradas no meio ambiente, capazes de causar infec??es em diversas esp?cies de seres vivos, incluindo aves, su?nos e humanos. Tais infec??es podem ser assintom?ticas, clinicamente significantes e, em alguns casos, fatais. No entanto, existe a necessidade de se disponibilizarem t?cnicas que ofere?am uma identifica??o conclusiva de bact?rias estreitamente relacionadas. As t?cnicas de biologia molecular se apresentam como ferramentas promissoras para uma identifica??o mais precisa. MATERIAIS E M?TODOS: Neste trabalho foram avaliados os marcadores moleculares RNAr 16S, hsp65 e rpoB aplicados ? distin??o de membros do MAC isolados no Laborat?rio de Micobact?rias do Instituto Evandro Chagas. RESULTADOS: Amostras de MAC colhidas em 15 pacientes foram previamente avaliadas pelo m?todo de an?lise de polimorfismo de fragmentos de restri??o do gene hsp65 (PRA-hsp65), que forneceu tr?s diferentes perfis: (I) BstEII: 235/115/100, HaeIII:145/130/60; (II) BstEII:235/210, HaeIII:130/105; e (III) : 235/210, HaeIII:145/130. ?rvores filogen?ticas foram constru?das ap?s an?lise do RNAr 16S, em que as amostras foram distribu?das em dois grupos, semelhante ao encontrado na an?lise de hsp65. Entretanto, os resultados de rpoB foram discordantes daqueles das outras ?rvores, devido ? modifica??o de topologia. CONCLUS?O: Os achados deste estudo sugerem que diferentes for?as evolucion?rias podem estar atuando sobre o gene rpoB, e, desta forma, ? necess?rio que se tenha precau??o ao estabelecer esse alvo para fins taxon?micos. Adicionalmente, recomenda-se que mais de um marcador, incluindo o RNAr 16S, seja avaliado para a identifica??o micobacteriana. 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.

Centro Universit?rio do Par?. Curso de P?s-Gradua??o em An?lises Cl?nicas. Bel?m, PA, Brasil / Laborat?rio de Patologia Cl?nica e Laboratorial Dr. Paulo Azevedo. Bel?m, PA, Brasil. Universidade Federal do Par?. Departamento de Medicina. Bel?m, PA, Brasil. Centro Universit?rio do Par?. Curso de P?s-Gradua??o em An?lises Cl?nicas. Bel?m, PA, Brasil. Laborat?rio de Patologia Cl?nica e Laboratorial Dr. Paulo Azevedo. Bel?m, PA, Brasil. Laborat?rio de Patologia Cl?nica e Laboratorial Dr. Paulo Azevedo. Bel?m, PA, Brasil. Centro Universit?rio do Par?. Curso de P?s-Gradua??o em An?lises Cl?nicas. Bel?m, PA, Brasil / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. A Serratia marcescens tem sido relatada como importante agente de infec??es relacionadas ? sa?de, destacando-se por apresentar elevado n?vel de resist?ncia intr?nseca aos antimicrobianos usados em neonatologia, al?m de persistir por longos per?odos no ambiente hospitalar. Neste trabalho foram avaliadas, por m?todos fenot?picos e moleculares, S. marcescens recuperadas a partir de coloniza??o do trato gastrointestinal ou sepse tardia em neonatos internados em Unidade Neonatal em Bel?m. A identifica??o das S. marcescens e o teste de sensibilidade foram realizados por meio de sistema automatizado Vitek (BioM?rieux); a suscetibilidade ao ertapenem foi avaliada com aux?lio de disco contendo 10 ?g da droga (Oxoide). A genotipagem foi feita por ERIC-PCR usando os primers ERIC1 (5'-TGAATCCCCAGGAGCTTACAT-3') e ERIC2 (5'-AAGTAAGTGACTGGGGTGAGCG-3'). Foram obtidas 22 cepas de S. marcescens, sendo 15 recuperadas de hemoculturas, e sete de vigil?ncia (swab retal); todas apresentaram resist?ncia a: ampicilina, ampicilina-sulbactam, gentamicina e cefalotina. N?o foi observada resist?ncia a: ciprofloxacina, imipenem, meropenem e ertapenem. Quanto aos demais antibi?ticos avaliados, o perfil de suscetibilidade foi vari?vel. Foram obtidos 11 padr?es de amplifica??o por ERIC-PCR, dois foram compartilhados por 14 isolados. Foi poss?vel observar um padr?o polim?rfico caracter?stico para as cepas provenientes de coloniza??o gastrointestinal, exceto em dois casos, que apresentaram padr?es genot?picos relacionadas a casos de sepse. Os dados obtidos neste trabalho confirmam o elevado ?ndice de resist?ncia da S. marcescens aos antimicrobianos; no entanto, todos os isolados apresentaram sensibilidade ? ciprofloxacina e aos carbapen?micos. A tipagem por meio de antibiograma e ERIC-PCR sugere dispers?o de clones associados ? coloniza??o ou sepse entre alas na Unidade Neonatal do hospital estudado. Serratia marcescens has been reported as an important agent of health care-related infections and has been highlighted for presenting a high level of intrinsic resistance to antimicrobials used in neonatology, besides persisting in hospital environments for long periods. In this work, S. marcescens was recovered from colonies in the gastrointestinal tract or late sepsis in newborn infants hospitalized in a Neonatal Unit in Bel?m. The identification of S. marcescens and the sensitivity test was carried out using a Vitek (BioM?rieux) automated system; susceptibility to ertapenem was assessed using e-test strips (Oxoid). Genotyping was executed by ERIC-PCR using the primers ERIC1 (5?-TGAATCCCCAGGAGCTTACAT-3?) and ERIC2 (5?-AAGTAAGTGACTGGGGTGAGCG-3?). Twenty-two strains of S. marcescens were recovered: 15 from hemocultures and seven from surveillance (rectal swab culture). All presented resistance to ampicillin, ampicillinsulbactam, gentamicin and cephalothin. There were no indications of resistance to ciprofloxacin, imipenem, meropenem or ertapenem. The susceptibility profiles varied for other antibiotics. Eleven amplification patterns by ERIC-PCR were obtained, and two were shared by 14 isolates. It was possible to observe a characteristic polymorphic pattern in the strains from gastrointestinal colonization, except for two cases, which presented genotypic patterns related to cases of sepsis. The data obtained in this work confirm the high level of resistance of S. marcescens against antimicrobials; however, all isolates displayed sensitivity to ciprofloxacin and carbapenemics. Antibiogram and ERIC-PCR typing suggest a dispersion of clones associated with colonization or sepsis among the wards of the Neonatal Unit in the surveyed hospital.

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Biologia Molecular. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Biologia Molecular. Ananindeua, PA, Brasil / Universidade do Estado do Par?. Centro de Ci?ncias Sociais e Educa??o. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Biologia Molecular. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Biologia Molecular. Ananindeua, PA, Brasil. INTRODU??O: As Escherichia coli diarreiog?nicas (DEC) s?o consideradas importante causa de diarr?ia nos pa?ses em desenvolvimento. Para sua correta identifica??o, esses microrganismos devem ser diferenciados dos membros n?o patog?nicos da microbiota intestinal. As DEC podem ser classificadas em seis categorias, de acordo com seu mecanismo de patogenicidade. MATERIAIS E M?TODOS: Foram desenvolvidos e avaliados dois sistemas de PCR multiplex para a detec??o de E. coli enteropatog?nicas (EPEC), enterotoxig?nica (ETEC), enteroinvasora (EIEC) e produtora da toxina de Shiga (STEC). RESULTADOS: Quatro categorias diarreiog?nicas foram detectadas entre os isolados de E. coli obtidos de indiv?duos infectados pelo HIV, com EIEC e EPEC representando os pat?tipos mais frequentes. Duas STEC stx1 e eae positivas foram identificadas, sendo este o primeiro relato de isolamento dessa categoria em indiv?duos infectados pelo HIV no estado do Par?. A PCR multiplex mostrou-se como t?cnica eficiente, r?pida e reprodut?vel para detec??o dos isolados de DEC. Os dois sistemas de PCR multiplex apresentaram resultados 100% concordantes com os encontrados na PCR simples. CONCLUS?O: Devido sua simplicidade, economia e efici?ncia torna-se poss?vel a aplica??o dos protocolos multiplex para agilizar o diagn?stico molecular das categorias de DEC, o que promover? a elabora??o de novos projetos de pesquisa e dar?o suporte as atividades de vigil?ncia epidemiol?gica desenvolvidos pelos institutos de sa?de p?blica. 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 stx1- and eae- positive were identified. This is the first report of this kind of isolation in individuals infected with HIV in the State of Par?. 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.