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To identify clinical and therapeutic features of pulmonary nontuberculous mycobacterial (PNTM) disease, we conducted a retrospective analysis of patients referred to the Brazilian reference center, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil, who received a diagnosis of PNTM during 1993–2011 with at least 1 respiratory culture positive for NTM. Associated conditions included bronchiectasis (21.8%), chronic obstructive pulmonary disease (20.7%), cardiovascular disease (15.5%), AIDS (9.8%), diabetes (9.8%), and hepatitis C (4.6%).Two patients had Hansen disease; 1 had Marfan syndrome. Four mycobacterial species comprised 85.6% of NTM infections: Mycobacterium kansasii, 59 cases (33.9%); M. avium complex, 53 (30.4%); M. abscessus, 23 (13.2%); and M. fortuitum, 14 (8.0%). A total of 42 (24.1%) cases were associated with rapidly growing mycobacteria. In countries with a high prevalence of tuberculosis, PNTM is likely misdiagnosed as tuberculosis, thus showing the need for improved capacity to diagnose mycobacterial disease as well as greater awareness of PNTM disease prevalence.

Nontuberculous mycobacterial infections can cause destructive tenosynovitis of the hand. We report on and discuss the clinical course and distinctive radiologic findings of two patients with hand tenosynovitis secondary to M. marinum and intracellulare infection, which are different from those of the nontuberculous mycobacterial infections reported in the previous literature.

Objective: One hundred thirty-one cases of postsurgical infections were reported in Southern Region of Brazil between August 2007 and January 2008. Thirty-nine (29.8%) cases were studied; this report describes epidemiological findings, species identification, antimicrobial susceptibility and clonal diversity of rapidly growing mycobacteria isolated in this outbreak. Methods: All 39 isolates were analyzed by Ziehl-Nielsen stained smear, bacterial culture and submitted to rpoB partial gene sequencing for identification. The isolates were also evaluated for their susceptibility to amikacin, cefoxitin, clarithromycin, ciprofloxacin, doxycycline, tobramycin and sulfamethoxazole. Results: Thirty-six isolates out of the confirmed cases were identified as Mycobacterium massiliense and the remaining three were identified as Mycobacterium abscessus, Mycobacterium chelonae and Mycobacterium fortuitum. All M. massiliense isolates were susceptible to amikacin (MIC90 = 8 μg/mL) and clarithromycin (MIC90 = 0.25 μg/mL) but resistant to cefoxitin, ciprofloxacin, doxycycline, tobramycin and sulfamethoxazole. Molecular analysis by pulsed-field gel electrophoresis clustered all 36 M. massiliense isolates and showed the same pattern (BRA 100) observed in three other outbreaks previously reported in Brazil. Conclusions: These findings suggest a common source of infection for all patients and reinforce the hypotheses of spread of M. massiliense BRA100 in Brazilian hospital surgical environment in recent years. Keywords: mycobacteria, atypical, mycobacterium infections, microbiological analysis

Infecções por micobactéria não tuberculosa (MNT) representam uma emergência epidemiológica e sanitária, especialmente, em pacientes submetidos a procedimentos invasivos. Frente ao exposto, objetivou-se analisar as evidências científicas, na literatura científica, sobre a ocorrência no Brasil de infecções por MNT em pacientes cirúrgicos. Utilizou-se como método de pesquisa a revisão integrativa da literatura nas bases de dados Lilacs, Medline/Pubmed, ISI Web of Science e Biblioteca Cochrane. Foram selecionadas 15 publicações sobre a temática nos últimos 30 anos que estavam direcionadas às medidas de prevenção e controle com foco na vigilância pós-alta, no uso de antibioticoterapia e glutaraldeído. Cirurgias oftalmológicas, estéticas, cardíacas e procedimentos laparoscópicos e artroscópicos foram as mais investigadas. A situação nacional das MNTs é preocupante, ainda mais quando se reconhece a possibilidade de subnotificação.Las infecciones por micobacteria no tuberculosa (MNT) representan una emergencia epidemiológica y sanitaria, especialmente, en pacientes sometidos a procedimientos invasivos. Frente a lo expuesto, se tuvo como objetivo analizar las evidencias científicas, en la literatura científica, sobre la ocurrencia en el Brasil de infecciones por MNT en pacientes quirúrgicos. Se utilizó como método de investigación la revisión integrativa de la literatura en las bases de datos Lilacs, Medline/Pubmed, ISI Web of Science y Biblioteca Cochrane. Se seleccionaron 15 publicaciones sobre la temática en los últimos 30 años que estaban orientadas a las medidas de prevención y control con foco en la vigilancia post alta, en el uso de antibioticoterapia y glutaraldehido. Cirugías oftalmológicas, estéticas, cardíacas y procedimientos laparoscópicos y artroscópicos fueron las más investigadas. La situación nacional de las MNTs es preocupante, aun más cuando se reconoce la posibilidad de subnotificación.Infections caused by nontuberculous mycobacteria (MNT) represent an epidemiological and health emergency, especially in patients undergoing invasive procedures. Based on these, we aimed to analyze the scientific evidence, the scientific literature, on the occurrence in Brazil of MNT infections in surgical patients. We used as a research method integrative review of the literature using the databases Lilacs, Medline/Pubmed, ISI Web of Science and the Cochrane Library. We selected 15 publications on this theme from the last 30 years that were directed at methods of prevention and control, with a focus on post-discharge surveillance, the use of antibiotics and glutaraldehyde. Eye surgery, cosmetic, heart, laparoscopic and arthroscopic procedures were the most commonly investigated. The national situation of MNTs is concerning, especially when one recognizes the possibility of underreporting.

Infecções por micobactéria não tuberculosa (MNT) representam uma emergência epidemiológica e sanitária, especialmente, em pacientes submetidos a procedimentos invasivos. Frente ao exposto, objetivou-se analisar as evidências científicas, na literatura científica, sobre a ocorrência no Brasil de infecções por MNT em pacientes cirúrgicos. Utilizou-se como método de pesquisa a revisão integrativa da literatura nas bases de dados Lilacs, Medline/Pubmed, ISI Web of Science e Biblioteca Cochrane. Foram selecionadas 15 publicações sobre a temática nos últimos 30 anos que estavam direcionadas às medidas de prevenção e controle com foco na vigilância pós-alta, no uso de antibioticoterapia e glutaraldeído. Cirurgias oftalmológicas, estéticas, cardíacas e procedimentos laparoscópicos e artroscópicos foram as mais investigadas. A situação nacional das MNTs é preocupante, ainda mais quando se reconhece a possibilidade de subnotificação. Infections caused by nontuberculous mycobacteria (MNT) represent an epidemiological and health emergency, especially in patients undergoing invasive procedures. Based on these, we aimed to analyze the scientific evidence, the scientific literature, on the occurrence in Brazil of MNT infections in surgical patients. We used as a research method integrative review of the literature using the databases Lilacs, Medline/Pubmed, ISI Web of Science and the Cochrane Library. We selected 15 publications on this theme from the last 30 years that were directed at methods of prevention and control, with a focus on post-discharge surveillance, the use of antibiotics and glutaraldehyde. Eye surgery, cosmetic, heart, laparoscopic and arthroscopic procedures were the most commonly investigated. The national situation of MNTs is concerning, especially when one recognizes the possibility of underreporting. Las infecciones por micobacteria no tuberculosa (MNT) representan una emergencia epidemiológica y sanitaria, especialmente, en pacientes sometidos a procedimientos invasivos. Frente a lo expuesto, se tuvo como objetivo analizar las evidencias científicas, en la literatura científica, sobre la ocurrencia en el Brasil de infecciones por MNT en pacientes quirúrgicos. Se utilizó como método de investigación la revisión integrativa de la literatura en las bases de datos Lilacs, Medline/Pubmed, ISI Web of Science y Biblioteca Cochrane. Se seleccionaron 15 publicaciones sobre la temática en los últimos 30 años que estaban orientadas a las medidas de prevención y control con foco en la vigilancia post alta, en el uso de antibioticoterapia y glutaraldehido. Cirugías oftalmológicas, estéticas, cardíacas y procedimientos laparoscópicos y artroscópicos fueron las más investigadas. La situación nacional de las MNTs es preocupante, aun más cuando se reconoce la posibilidad de subnotificación.

SummaryRecent studies suggest an increasing prevalence of pulmonary nontuberculous mycobacteria (NTM) disease. In the absence of prevalence and cost data, the public health burden of pulmonary NTM disease is difficult to assess. The goal of this study was to assess costs associated with NTM disease treatment and to identify risk factors associated with increased costs. Records from subjects with pulmonary NTM disease enrolled in a natural history protocol were abstracted for presenting symptoms, comorbidities, microbiology, and treatment histories. Antibiotic frequency, duration, adverse reaction, and costs were noted, the total antibiotic burden and cost were calculated, and risk factors associated with high costs were analyzed. From Jan 2004 to Dec 2005, 33 subjects were enrolled; 27 met disease criteria and had sufficient data to assess antibiotic use. Mycobacterium avium complex was present in 89% and Mycobacterium abscessus was present in 21% of subjects. Subjects received a median of 5 (1–10) antibiotics. Adverse effects were common seen in up to 50% with common antibiotics and up to 100% with uncommonly used antibiotics. Median burden of treatment was 2638 (84–7689) drug-days and the median total cost per patient was $19,876 ($398–70,917). Subjects with high treatment costs had an adjusted 9.5 fold (95% CI 1.5–97.2) likelihood of having M. abscessus and a 4.2 fold (95% CI 0.6–59.3) increased likelihood of having more extensive disease. Pulmonary NTM represent an underappreciated disease burden in the US population, with an associated treatment cost comparable to that for other chronic diseases of infectious origin such as HIV/AIDS.

To report the presence of viable mycobacteria in a patient with keratitis treated for 6 months. Species identification was performed using the PRA method (polymerase chain reaction followed by restriction endonuclease analysis). Clonality was evaluated with RAPD (randomly amplified polymorphic DNA) and ERIC-PCR (enterobacterial repetitive intergenic consensus - polymerase chain reaction) methods. The patient reported trauma due to a metallic foreign body 3 weeks prior to presentation. Initial corneal scraping cultures revealed Mycobacterium abscessus. After 6 months of topical and systemic treatment the patient presented with no active inflammation and was considered clinically cured. An optic penetrating keratoplasty was performed. Culture of the excised cornea revealed Mycobacterium abscessus. Both isolates had the same clonal origin. The most interesting finding of this case report was the positive culture of the excised cornea after 6 months of intensive specific topical therapy. To our knowledge, this is the first report in the literature showing this possibility in the treatment of Mycobacterial keratitis. Thus, Mycobacterium abscessus may present viable bacteria after long-term treatment and should be followed carefully for a long period of time after tapering the medication.O objetivo do caso é descrever a presença de micobactérias viáveis em pacientes com ceratite, 6 meses após tratamento intensivo. A identificação de espécies, foi efetuada usando método PRA (polymerase chain reaction seguida pela restriction endonuclease analysis). Clonalidade foi avaliada pelos métodos RAPD (randomly amplified polymorphic DNA) e ERIC-PCR (enterobacterial repetitive intergenic consensus - polymerase chain reaction). Paciente refere trauma com corpo estranho metálico há 3 semanas. A cultura da córnea revelou Mycobacterium abscessus. Após 6 meses de tratamento tópico e sistêmico, paciente apresentava-se sem inflamação, sendo considerado clinicamente curado. Realizou-se então, uma ceratoplastia penetrante com intuitos ópticos. A cultura da córnea transplantada revelou micobactérias de mesma origem clonal. O achado mais interessante neste relato, foi a positividade da cultura da córnea transplantada após 6 meses de intenso tratamento específico. Ao nosso conhecimento, esse é o primeiro caso relatado na literatura mostrando essa possibilidade em tratamento de ceratites por micobactérias. Assim, os pacientes com ceratite por Mycobacterium abscessus podem apresentar bactérias viáveis após longo tempo de tratamento específico e precisam ser seguidos cuidadosamente por um longo período de tempo.

Atypical fast-growing Mycobacterium species are usually identified after laser-assisted in situ keratomileusis, cosmetic surgeries, and catheter-related, pulmonary or soft tissue infections. We herein present the case of a 56-year-old man with purulent discharge, redness, and foreign body sensation in his left eye. He underwent two surgeries that partially controlled the infection but were not curative. Corneal transplantation was performed, and a biopsy of the excised cornea indicated Mycobacterium aurum infection, which was confirmed by polymerase chain reaction-restriction fragment length polymorphism analysis. This appears to be the first documented case of keratitis attributable to the non-tuberculous mycobateria M. aurum. The intractable extra-ocular progression of the disease in the absence of general signs or symptoms was notable. We suggest considering non-tuberculous mycobacteria among the probable causes of complicated keratitis or keratitis that does not respond to drug treatment, especially in regions where tuberculosis is endemic.

Atypical fast-growing Mycobacterium species are usually identified after laser-assisted in situ keratomileusis, cosmetic surgeries, and catheter-related, pulmonary or soft tissue infections. We herein present the case of a 56-year-old man with purulent discharge, redness, and foreign body sensation in his left eye. He underwent two surgeries that partially controlled the infection but were not curative. Corneal transplantation was performed, and a biopsy of the excised cornea indicated Mycobacterium aurum infection, which was confirmed by polymerase chain reaction-restriction fragment length polymorphism analysis. This appears to be the first documented case of keratitis attributable to the non-tuberculous mycobateria M. aurum. The intractable extra-ocular progression of the disease in the absence of general signs or symptoms was notable. We suggest considering non-tuberculous mycobacteria among the probable causes of complicated keratitis or keratitis that does not respond to drug treatment, especially in regions where tuberculosis is endemic. Keywords: Mycobacteria, atypical, Keratitis, Corneal transplantation

OBJETIVO: Identificar as espécies de micobactérias encontradas no escarro de pacientes com suspeita de tuberculose pulmonar e analisar o impacto dessas identificações na abordagem terapêutica. MÉTODOS: Foram avaliados 106 pacientes com suspeita de tuberculose pulmonar encaminhados para o serviço de pneumologia de um hospital público em Teresina, Piauí. Espécimes de escarro matinal foram avaliados quanto à presença de micobactérias por baciloscopia e cultura. Foram utilizadas PCR e análise de restrição enzimática do gene hsp65 (PRA-hsp65) para a identificação das cepas de micobactérias isoladas em cultura. RESULTADOS: Foram analisadas 206 amostras de escarro. A idade dos pacientes variou de 15 a 87 anos, sendo 67% do gênero masculino. Tosse ocorreu em 100% dos casos. O padrão radiográfico predominante foi de lesão moderada, observada em 70%. A positividade no esfregaço foi de 76%, e isolamento em cultura ocorreu em 91% das culturas executadas. Testes tradicionais identificaram micobactérias não tuberculosas (MNT) em 9% dos isolados. O método PRA-hsp65 confirmou esses dados, mostrando sete padrões de bandas capazes de identificar as espécies de MNT isoladas: Mycobacterium kansasii; M. abscessus 1; M. abscessus 2; M. smegmatis; M. flavescens 1; M. gordonae 5 e M. gordonae 7. Todos os pacientes com MNT tinham mais de 60 anos, e observaram-se bronquiectasias em 88% das radiografias. Houve dois casos de reinfecção, identificados inicialmente como infecção por M. abscessus e M. kansasii. CONCLUSÕES: As MNT causam infecção pulmonar em pacientes imunocompetentes, e a identificação das MNT é importante para estabelecer o diagnóstico correto e a decisão terapêutica mais adequada. O método PRA-hsp65 é útil para identificar espécies de MNT e pode ser implantado em laboratórios de biologia molecular não especializados em micobactérias. OBJECTIVE: To identify mycobacterial species in the sputum of patients suspected of having pulmonary tuberculosis and to determine the impact that the acquisition of this knowledge has on the therapeutic approach. METHODS: We evaluated 106 patients suspected of having pulmonary tuberculosis and referred to the pulmonology department of a public hospital in the city of Teresina, Brazil. Morning sputum specimens were evaluated for the presence of mycobacteria by sputum smear microscopy and culture. We used PCR and restriction enzyme analysis of the hsp65 gene (PRA-hsp65) to identify the strains of mycobacteria isolated in culture. RESULTS: A total of 206 sputum samples were analyzed. Patient ages ranged from 15 to 87 years, and 67% were male. There was cough in 100% of the cases. The predominant radiographic pattern was moderate disease, observed in 70%. Smear positivity was 76%, and isolation in culture occurred in 91% of the cultures. Traditional tests identified nontuberculous mycobacteria (NTM) in 9% of the isolates. The PRA-hsp65 method confirmed these data, showing seven band patterns that were able to identify the isolated species of NTM: Mycobacterium kansasii; M. abscessus 1; M. abscessus 2; M. smegmatis; M. flavescens 1; M. gordonae 5; and M. gordonae 7. All of the patients with NTM were over 60 years of age, and bronchiectasis was seen in 88% of the X-rays. There were two cases of reinfection, initially attributed to M. abscessus and M. kansasii. CONCLUSIONS: In immunocompetent patients, NTM can infect the lungs. It is important to identify the specific NTM in order to establish the correct diagnosis and choose the most appropriate therapeutic regimen. The PRA-hsp65 method is useful in identifying NTM species and can be implemented in molecular biology laboratories that do not specialize in the identification of mycobacteria.

INTRODUCTION: Reports of infections caused by rapidly growing mycobacteria during plastic surgery have increased in recent years despite improvements in techniques of asepsis/antisepsis and antibiotic prophylaxis. Infections occurring after the insertion of breast implants are a cause of patient morbidity and a significant problem for the surgeon. METHODS: Breast implant surgery cases complicated by mycobacterial infections at the Infirmary ward 38th of the Santa Casa da Misericórdia, Rio de Janeiro were retrospectively reviewed. A description of the current guidelines for the prevention and treatment of mycobacteriosis is included. Laboratory confirmed and clinically suspected cases were included in this study. RESULTS: Of 483 augmentation mammaplasty cases, 3 patients developed mycobacterial infections in the last 3 years. In 2 patients, there was a suspicion of infection that was not confirmed by laboratory data. CONCLUSIONS: Prophylaxis is fundamental for reducing the incidence of mycobacteriosis during plastic surgery procedures. However, the identification, diagnosis, and treatment of mycobacterial diseases are important to minimize the morbidity of this type of infection INTRODUÇÃO: Nos últimos anos, foram crescentes os registros de infecções por micobactéria de crescimento rápido em cirurgia plástica, mesmo com a melhoria dos métodos de assepsia/ antissepsia e da antibioticoprofilaxia. A infecção após inclusão de implantes mamários causa grande morbidade às pacientes e transtorno ao cirurgião. MÉTODO: Estudo retrospectivo dos casos de infecção por micobactéria de crescimento rápido da 38ª Enfermaria da Santa Casa da Misericórdia do Rio de Janeiro, após inclusão de implantes mamários, em que são apresentadas propostas de prevenção e tratamento da micobacteriose. Foram incluídos os casos confirmados laboratorialmente e os clinicamente suspeitos. RESULTADOS: Até o presente momento foram confirmados 3 casos de infecção por micobactéria, num total de 483 mamaplastia de aumento no decorrer de 3 anos. Em 2 pacientes, houve suspeita de infecção, porém sem confirmação laboratorial. CONCLUSÕES: A profilaxia é o pilar fundamental para a redução do impacto da micobacteriose em procedimentos de cirurgia plástica. Entretanto, saber identificar, diagnosticar e tratar corretamente a micobacteriose é de suma importância para minimizar a morbidade da paciente

OBJETIVO: Identificar micobactérias não tuberculosas (MNT) isoladas de sítios estéreis em pacientes internados no Hospital Universitário Clementino Fraga Filho, Rio de Janeiro (RJ) entre 2001 e 2006. MÉTODOS: Durante o período do estudo, 34 isolados de MNT de sítios estéreis de 14 pacientes, a maioria HIV positivos, foram submetidos a identificação fenotípica e hsp65 PCR-restriction enzyme analysis (PRA, análise por enzimas de restrição por PCR do gene hsp65). RESULTADOS: A maioria dos isolados foi identificada como Mycobacterium avium, seguida por M. monacense, M. kansasii e M. abscessus em menores proporções. CONCLUSÕES: A combinação de PRA, um método relativamente simples e de baixo custo, com algumas características fenotípicas pode fornecer a identificação correta de MNT na rotina de laboratórios clínicos. OBJECTIVE: To identify nontuberculous mycobacteria (NTM) isolated from sterile sites in patients hospitalized between 2001 and 2006 at the Clementino Fraga Filho University Hospital, located in the city of Rio de Janeiro, Brazil. METHODS: During the study period, 34 NTM isolates from sterile sites of 14 patients, most of whom were HIV-positive, were submitted to phenotypic identification and hsp65 PCR-restriction enzyme analysis (PRA). RESULTS: Most isolates were identified as Mycobacterium avium, followed by M. monacense, M. kansasii, and M. abscessus. CONCLUSIONS: The combination of PRA, a relatively simple and inexpensive method, with the evaluation of a few phenotypic characteristics can allow NTM to be accurately identified in the routine of clinical laboratories.

OBJETIVO: Estudar a ocorrência de micobactérias não-tuberculosas e a variabilidade das espécies isoladas na região atendida pelo Instituto Adolfo Lutz-Regional de São José do Rio Preto-no período entre 1996 e 2005, assim como mostrar a importância do diagnóstico laboratorial. MÉTODOS: A partir de amostras pulmonares e extrapulmonares, foi realizado o isolamento de micobactérias, e estas foram identificadas por métodos fenotípicos e pelo método molecular polymerase chain reaction-restriction enzyme analysis. RESULTADOS: Foram isoladas 317 cepas de micobactérias não-tuberculosas: complexo Mycobacterium avium, 182 (57,4%); M. gordonae, 33 (10,4%); M. fortuitum, 25(7,9%); M. chelonae, 8 (2,5%); complexo M. terrae, 8 (2,5%); M. kansasii, 7 (2,2%); e espécies menos freqüentes, 54 (17%). No período, foram caracterizados 72 casos (33,3%) de micobacterioses, de acordo com os critérios bacteriológicos estabelecidos pela American Thoracic Society (2007).Desses, complexo M. avium foi responsável por 56 casos, sendo que 29 (51,8%) foram caracterizados como doença disseminada. M. fortuitum foi responsável por 6 casos; M. gordonae, 3; M. chelonae, 2; M. abscessus, 1; M. kansasii, 1; M. intracellulare, 1; M. malmoense, 1; e Mycobacterium ssp., 1. CONCLUSÕES: Os resultados obtidos mostraram a importância do diagnóstico bacteriológico das micobacterioses, pois a identificação das espécies possibilita a introdução de um tratamento adequado precocemente. OBJECTIVE: To study the incidence of nontuberculous mycobacteria and the range of species isolated between 1996 and 2005 at a regional branch of the Adolfo Lutz Institute-located in the city of São José do Rio Preto, Brazil-and to show the importance of laboratory testing. METHODS: Mycobacteria were isolated from pulmonary and extrapulmonary specimens and identified through phenotyping and molecular methods (polymerase chain reaction-restriction enzyme analysis). RESULTS: We isolated 317 nontuberculous mycobacterium strains: Mycobacterium avium complex, 182 (57.4%); M. gordonae, 33 (10.4%); M. fortuitum, 25 (7.9%); M. chelonae, 8 (2.5%); M. terrae complex, 8(2.5%); M.kansasii, 7 (2.2%); and less frequent species, 54 (17%). During this period, 72 cases (33.3%) were characterized as mycobacteriosis, according to bacteriological criteria established by the American Thoracic Society in 2007. Of those 72 cases, 56 were attributed to M.avium complex. Of those 56, 29 (51.8%) were characterized as disseminated disease. Six cases were attributed to M. fortuitum, 3 to M. gordonae, 2 to M. chelonae, 1 to M. abscessus, 1 to M. kansasii, 1 to M. intracellulare, 1 to M. malmoense and 1 to Mycobacterium ssp. CONCLUSIONS: These results show the importance of the bacteriological diagnosis, since identification of the species enables early and appropriate treatment.

OBJETIVO: A micobacteriose é frequentemente diagnosticada entre pacientes infectados pelo HIV. Em Moçambique, onde apenas um pequeno número de pacientes encontra-se sob tratamento anti-retroviral, e a tuberculose tem alta prevalência, existe a necessidade de melhor caracterização destes agentes bacterianos, em nível de espécie, bem como de se caracterizar os padrões de resistência às drogas antituberculosas. MÉTODOS: Em uma coorte de 503 indivíduos HIV positivos suspeitos de tuberculose pulmonar, 320 apresentaram positividade para baciloscopia ou cultura no escarro e no lavado brônquico. RESULTADOS: Bacilos álcool-ácido resistentes foram detectados no escarro em 73% dos casos com cultura positiva. De 277 isolados em cultura, apenas 3 mostraram-se tratar de micobactérias não-tuberculosas: 2 Mycobacterium avium e uma M. simiae. Todos os isolados de M. tuberculosis inicialmente caracterizados através de reação em cadeia de polimerase (RCP) do gene hsp65 foram posteriormente caracterizados como tal através de RCP do gene gyrB. Resistência à isoniazida foi encontrada em 14% dos casos; à rifampicina em 6%; e multirresistência em 5%. Pacientes previamente tratados para tuberculose mostraram tendência a taxas maiores de resistência às drogas de primeira linha. O padrão radiológico mais freqüente encontrado foi o infiltrado intersticial (67%), seguido da presença de linfonodos mediastinais (30%), bronquiectasias (28%), padrão miliar (18%) e cavidades (12%). Os pacientes infectados por micobactérias não-tuberculosas não apresentaram manifestações clínicas distintas das apresentadas pelos outros pacientes. A mediana de linfócitos CD4 entre todos os pacientes foi de 134 células/mm³. CONCLUSÕES: Tuberculose e AIDS em Moçambique estão fortemente associadas, como é de se esperar em países com alta prevalência de tuberculose. Embora as taxas de resistência a drogas sejam altas, o esquema isoniazida-rifampicina continua sendo a escolha apropriada para o início do tratamento. OBJECTIVE: Mycobacteriosis is frequently diagnosed among HIV-infected patients. In Mozambique, where few patients are under antiretroviral therapy and the prevalence of tuberculosis is high, there is need for better characterization of mycobacteria at the species level, as well as for the identification of patterns of resistance to antituberculous drugs. METHODS: We studied a sample of 503 HIV-infected individuals suspected of having pulmonary tuberculosis. Of those 503, 320 tested positive for mycobacteria through sputum smear microscopy or culture of bronchoalveolar lavage fluid. RESULTS: Acid-fast bacilli were observed in the sputum of 73% of the individuals presenting positive cultures. Of 277 isolates tested, only 3 were nontuberculous mycobacteria: 2 were identified as Mycobacterium avium and one was identified as M. simiae. Strains initially characterized as M. tuberculosis complex through polymerase chain reaction restriction analysis (PRA) of the hsp65 gene were later confirmed as such through PRA of the gyrB gene. Among the M. tuberculosis isolates, resistance patterns were as follows: to isoniazid, 14%; to rifampin, 6%; and multidrug resistance, 5%. Previously treated cases showed significantly higher rates of resistance to first-line antituberculous drugs. The most common radiological pattern was interstitial infiltrate (in 67%), followed by mediastinal lymph node enlargement (in 30%), bronchiectasis (in 28%), miliary nodules (in 18%) and cavitation (in 12%). Patients infected with nontuberculous mycobacteria presented clinical profiles indistinguishable from those of other patients. The median CD4 lymphocyte count in this group was 134 cells/mm³. CONCLUSIONS: There is a strong association between tuberculosis and AIDS in Mozambique, as expected in a country with a high prevalence of tuberculosis. Although drug resistance rates are high, the isoniazid-rifampin regimen continues to be the appropriate choice for initial therapy.

OBJETIVO: Este estudo teve por objetivo descrever a freqüência das espécies de micobactérias não tuberculosas (MNT) identificadas laboratorialmente a partir do isolamento de sítios não estéreis (escarro) de indivíduos infectados ou não pelo vírus HIV na Baixada Santista (SP), período de 2000 a 2005. MÉTODOS: Foi realizada análise retrospectiva dos resultados de baciloscopia e cultura, disponíveis nos registros do laboratório regional de tuberculose, Instituto Adolfo Lutz-Santos. RESULTADOS: Analisou-se 194 cepas de MNT correspondentes a 125 indivíduos, sendo 73 (58,4%) HIV negativos e 52 (41,6%) HIV positivos. Foram identificadas 13 diferentes espécies: Mycobacterium kansasii; complexo M. avium; M. fortuitum; M. peregrinum; M. gordonae; M. terrae; M. nonchromogenicum; M. intracellulare; M. flavescens; M. bohemicum; M. chelonae; M. shimoidei; e M. lentiflavum. Em 19,2% dos casos obteve-se diagnóstico bacteriológico confirmado pelo isolamento da mesma espécie em no mínimo duas amostras consecutivas. CONCLUSÕES: Os resultados mostram a importância da realização sistemática da identificação de MNT na rotina laboratorial e sua integração com a clínica, podendo contribuir na caracterização da doença e ações de efetivo controle, como nas populações co-infectadas tuberculose e HIV. OBJECTIVE: The present study aims at describing the frequency of nontuberculous mycobacteria (NTM) species identified through laboratory testing of samples collected from non-sterile sites (sputum), as well as its frequency in HIV-infected and non-HIV-infected individuals in the Baixada Santista region of the state of São Paulo, Brazil, in the period from 2000 to 2005. METHODS: Retrospective analysis of sputum smear microscopy results and culture was conducted based on the records on file at the Instituto Adolfo Lutz-Santos, the regional tuberculosis laboratory. RESULTS: We analyzed 194 NTM strains isolated from 125 individuals, of whom 73 (58.4%) were HIV-negative and 52 (41.6%) were HIV-positive. Thirteen different species were identified: Mycobacterium kansasii; M. avium complex; M. fortuitum; M. peregrinum; M. gordonae; M. terrae; M. nonchromogenicum; M. intracellulare; M. flavescens; M. bohemicum; M. chelonae; M. shimoidei; and M. lentiflavum. In 19.2% of the cases, the bacteriological diagnosis was confirmed by isolation of the same species in at least two consecutive samples. CONCLUSIONS: Our results show the importance of including systematic identification of NTM in the laboratory routine, and that its integration into the clinical routine could improve the characterization of the disease, thereby informing the planning of effective control measures in specific populations, such as individuals presenting tuberculosis/HIV co-infection.

Although the treatment of pulmonary diseases due to nontuberculous mycobacteria (NTM) requires the long-term use of antibiotics in combination, the treatment success rates are unsatisfactory. We evaluated the clinical characteristics and surgical outcomes of 23 patients with NTM lung diseases who had underwent pulmonary resection. The median age of the patients was 45 yr. Of the 23 patients, 10 had Mycobacterium avium-intracellulare complex infection, 12 had M. abscessus infection, and one had M. xenopi infection. The indications for surgery were antibiotic therapy failure (n=11), remnant cavitary lesion with high probability of relapse (n=8), and massive hemoptysis (n=4). The most common procedure was lobectomy (48%). Postoperative complications occurred in eight patients (35%), including postoperative pneumonia (n=3) and late bronchopleural fistula (n=2). Negative sputum culture conversion was achieved and maintained in all except two mortalities. Although it is associated with a relatively high complication rate, patients with NTM lung disease whose disease is localized to one lung and who can tolerate resectional surgery might be considered for surgery, if there has been poor response to drug therapy or if the patients develop significant disease-related complications such as hemoptysis.

Atypische Mykobakterien sind ubiquitär vorkommende Bakterien, die vornehmlich in natürlichen und künstlichen Wasserquellen, wie zum Beispiel Leitungswasser, gefunden werden. Sie werden immer häufiger als Ursache unterschiedlicher Krankheitsbilder, insbesondere bei immunsupprimierten Patienten, gesehen. Um zu klären, ob Leitungswasser als Kontaminations- oder Infektionsquelle von Bedeutung ist, wurde es auf ubiquitäre Mykobakterien mittels quantitativer LightCycler Real-time PCR und Kultur untersucht. Dabei wurde sowohl eine Mycobacterium-gattungsspezifische PCR als auch eine M. xenopi-spezifische PCR eingesetzt.

Background: The frequency of diseases caused by non tuberculous mycobacteria has increased in the last years. Their clinical diagnosis is difficult, mainly in immunocompromised patients. The identification of these mycobacteria by traditional methods is based on phenotypic characteristics and the results are obtained two to four weeks after their isolation in primary cultures. Aim: To report a new identification method for non tuberculous mycobacteria. Material and methods: The restriction pattern analysis method was implemented. It is based on the amplification, using polymerase chain reaction (PCR), of a polymorphic region of 440 base pairs that codifies Hsp65 protein, followed by a digestion with BstE II and Hae III restriction enzymes. The results were compared with patterns established for each strain. Results: Sixty four strains of mycobacteria obtained from clinical samples and seven reference mycobacteria, were identified using the traditional methods and restriction pattern analysis. The latter method identified the same strain as the former in 87.5% of cases. In the remainder 12.5% of cases there was no agreement between both methods. In these, the sequencing of a fragment of a gene that codifies 16S ribosomal RNA, confirmed the correct identification by restriction patterns. Conclusions: Restriction pattern analysis is a rapid identification method for non tuberculous mycobaterial strains

Background: The frequency of diseases caused by non tuberculous mycobacteria has increased in the last years. Their clinical diagnosis is difficult, mainly in immunocompromised patients. The identification of these mycobacteria by traditional methods is based on phenotypic characteristics and the results are obtained two to four weeks after their isolation in primary cultures. Aim: To report a new identification method for non tuberculous mycobacteria. Material and methods: The restriction pattern analysis method was implemented. It is based on the amplification, using polymerase chain reaction (PCR), of a polymorphic region of 440 base pairs that codifies Hsp65 protein, followed by a digestion with BstE II and Hae III restriction enzymes. The results were compared with patterns established for each strain. Results: Sixty four strains of mycobacteria obtained from clinical samples and seven reference mycobacteria, were identified using the traditional methods and restriction pattern analysis. The latter method identified the same strain as the former in 87.5% of cases. In the remainder 12.5% of cases there was no agreement between both methods. In these, the sequencing of a fragment of a gene that codifies 16S ribosomal RNA, confirmed the correct identification by restriction patterns. Conclusions: Restriction pattern analysis is a rapid identification method for non tuberculous mycobaterial strains (Rev Med Chile 2006; 134: 868-73).

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:21:37Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:44:28Z : No. of bitstreams: 1 2-s2.0-24344448301.pdf: 16200 bytes, checksum: 9a42d316b1626dd185175d38498d9b51 (MD5) Made available in DSpace on 2014-05-27T11:21:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-09-13 Water samples (24 untreated water, 12 treated water and 24 served water) used in different stages of the slaughter process were examined to identify a possible source of pathogenic mycobacteria. The isolates were identified based on microscopy, morphological and biochemical features, mycolic acid analysis and molecular method - PCR-restriction-enzyme analysis. Eighteen mycobacterial strains were isolated from 60 water samples: 11 from untreated water, 5 from treated water and 2 from served water. All mycobacteria isolated were identified as Mycobacterium gordonae and showed the following PRA genotypes: III (27.8%), IV (38.9%) and V (33.3%). Faculdade de Cièncias Farmacêuticas UNESP, Rodovia Araraquara-Jaú Km 01, CEP 14801-902, Araraquara, SP Faculdade de Cièncias Agrárias UNICASTELO Campus VIII, Descalvado, SP Faculdade de Cièncias Farmacêuticas UNESP, Rodovia Araraquara-Jaú Km 01, CEP 14801-902, Araraquara, SP

O objetivo do caso é descrever a presença de micobactérias viáveis em pacientes com ceratite, 6 meses após tratamento intensivo. A identificação de espécies, foi efetuada usando método PRA (polymerase chain reaction seguida pela restriction endonuclease analysis). Clonalidade foi avaliada pelos métodos RAPD (randomly amplified polymorphic DNA) e ERIC-PCR (enterobacterial repetitive intergenic consensus - polymerase chain reaction). Paciente refere trauma com corpo estranho metálico há 3 semanas. A cultura da córnea revelou Mycobacterium abscessus. Após 6 meses de tratamento tópico e sistêmico, paciente apresentava-se sem inflamação, sendo considerado clinicamente curado. Realizou-se então, uma ceratoplastia penetrante com intuitos ópticos. A cultura da córnea transplantada revelou micobactérias de mesma origem clonal. O achado mais interessante neste relato, foi a positividade da cultura da córnea transplantada após 6 meses de intenso tratamento específico. Ao nosso conhecimento, esse é o primeiro caso relatado na literatura mostrando essa possibilidade em tratamento de ceratites por micobactérias. Assim, os pacientes com ceratite por Mycobacterium abscessus podem apresentar bactérias viáveis após longo tempo de tratamento específico e precisam ser seguidos cuidadosamente por um longo período de tempo. To report the presence of viable mycobacteria in a patient with keratitis treated for 6 months. Species identification was performed using the PRA method (polymerase chain reaction followed by restriction endonuclease analysis). Clonality was evaluated with RAPD (randomly amplified polymorphic DNA) and ERIC-PCR (enterobacterial repetitive intergenic consensus - polymerase chain reaction) methods. The patient reported trauma due to a metallic foreign body 3 weeks prior to presentation. Initial corneal scraping cultures revealed Mycobacterium abscessus. After 6 months of topical and systemic treatment the patient presented with no active inflammation and was considered clinically cured. An optic penetrating keratoplasty was performed. Culture of the excised cornea revealed Mycobacterium abscessus. Both isolates had the same clonal origin. The most interesting finding of this case report was the positive culture of the excised cornea after 6 months of intensive specific topical therapy. To our knowledge, this is the first report in the literature showing this possibility in the treatment of Mycobacterial keratitis. Thus, Mycobacterium abscessus may present viable bacteria after long-term treatment and should be followed carefully for a long period of time after tapering the medication. Universidade Federal de São Paulo (UNIFESP) UNIFESP Setor de Córnea e Doenças Externas Laboratório Fleury UNIFESP Departamento de Microbiologia, Imunologia e Parasitologia UNIFESP Departamento de Oftalmologia UNIFESP, Setor de Córnea e Doenças Externas UNIFESP, Depto. de Microbiologia, Imunologia e Parasitologia UNIFESP, Depto. de Oftalmologia SciELO

To report the presence of viable mycobacteria in a patient with keratitis treated for 6 months. Species identification was performed using the PRA method (polymerase chain reaction followed by restriction endonuclease analysis). Clonality was evaluated with RAPD (randomly amplified polymorphic DNA) and ERIC-PCR (enterobacterial repetitive intergenic consensus - polymerase chain reaction) methods. The patient reported trauma due to a metallic foreign body 3 weeks prior to presentation. Initial corneal scraping cultures revealed Mycobacterium abscessus. After 6 months of topical and systemic treatment the patient presented with no active inflammation and was considered clinically cured. An optic penetrating keratoplasty was performed. Culture of the excised cornea revealed Mycobacterium abscessus. Both isolates had the same clonal origin. The most interesting finding of this case report was the positive culture of the excised cornea after 6 months of intensive specific topical therapy. To our knowledge, this is the first report in the literature showing this possibility in the treatment of Mycobacterial keratitis. Thus, Mycobacterium abscessus may present viable bacteria after long-term treatment and should be followed carefully for a long period of time after tapering the medication. O objetivo do caso é descrever a presença de micobactérias viáveis em pacientes com ceratite, 6 meses após tratamento intensivo. A identificação de espécies, foi efetuada usando método PRA (polymerase chain reaction seguida pela restriction endonuclease analysis). Clonalidade foi avaliada pelos métodos RAPD (randomly amplified polymorphic DNA) e ERIC-PCR (enterobacterial repetitive intergenic consensus - polymerase chain reaction). Paciente refere trauma com corpo estranho metálico há 3 semanas. A cultura da córnea revelou Mycobacterium abscessus. Após 6 meses de tratamento tópico e sistêmico, paciente apresentava-se sem inflamação, sendo considerado clinicamente curado. Realizou-se então, uma ceratoplastia penetrante com intuitos ópticos. A cultura da córnea transplantada revelou micobactérias de mesma origem clonal. O achado mais interessante neste relato, foi a positividade da cultura da córnea transplantada após 6 meses de intenso tratamento específico. Ao nosso conhecimento, esse é o primeiro caso relatado na literatura mostrando essa possibilidade em tratamento de ceratites por micobactérias. Assim, os pacientes com ceratite por Mycobacterium abscessus podem apresentar bactérias viáveis após longo tempo de tratamento específico e precisam ser seguidos cuidadosamente por um longo período de tempo.

As the incidence of nontuberculous mycobacterial infection has been increasing recently in Korea, the importance of drug susceptibility test for clinical isolates of mycobacteria has become larger. In this study we determined the antimicrobial susceptibility patterns of clinical isolates of M. fortuitum and M. abscessus in Korea, and evaluated the efficacy of a modified broth microdilution method using 2,3-diphenyl-5-thienyl-(2)-tetrazolium chloride (STC), in terms of its ability to provide accurate and easy-to-read minimal inhibitory concentration (MIC) endpoints for the susceptibility testing of rapidly growing mycobacteria. Most isolates of M. fortuitum and M. abscessus in Korea are susceptible or intermediately susceptible to amikacin, cefoxitin, ciprofloxacin, and clarithromycin. Many isolates of M. fortuitum are susceptible to doxycycline, sulfamethoxazole, and imipenem, while many M. abscessus isolates are resistant to these drugs. In the present study, the modified broth microdilution method using STC was found to be reliable, easy to read, and inexpensive for M. fortuitum and M. abscessus susceptibility testing. The modified colorimetric MIC testing method using STC was proven to be a useful surrogate for RGM antibiotic susceptibility testing.

Mycobacterium kansasii is one of the most common cause of pulmonary diseases due to nontuberculous mycobacteria. We investigated the changing in the number of isolation of M. kansasii and the clinical characteristics of M. kansasii pulmonary disease in Korea. Through searching the database of the Korean Institute of Tuberculosis, we identified the cases of isolated M. kansasii from 1992 to 2002. The number of M. kansasii isolation had increased from once in 1992 to 62 in 2002. Fifteen patients with M. kansasii pulmonary disease were identified during the period January 1997 to December 2002. Twelve patients (80%) were male and fourteen (93%) were from highly industrialized areas. The most common symptom was a cough. Seven patients (47%) had a cavitary lesion and right upper lobe was most commonly involved. Patients responded well to isoniazid and rifampicin based regimens both bacteriologically and radiographically. In conclusion, M. kansasii isolation has increased, especially in highly industrialized areas, as well as other nontuberculous mycobacteria in Korea.

We aimed to compare the CT findings of nontuberculous mycobacterial pulmonary diseases caused by Mycobacterium avium-intracellulare complex (MAC) and Mycobacterium abscessus. Two chest radiologists analyzed retrospectively the thin-section CT findings of 51 patients with MAC and 36 with M. abscessus infection in terms of patterns and forms of lung lesions. No significant difference was found between MAC and M. abscessus infection in the presence of small nodules, tree-in-bud pattern, and bronchiectasis. However, lobar volume decrease (p=0.001), nodule (p=0.018), airspace consolidation (p=0.047) and thin-walled cavity (p=0.009) were more frequently observed in MAC infection. The upper lobe cavitary form was more frequent in the MAC (19 of 51 patients, 37%) group than M. abscessus (5 of 36, 14%) (p=0.029), whereas the nodular bronchiectatic form was more frequent in the M. abscessus group ([29 of 36, 81%] vs. [27 of 51, 53%] in MAC) (p=0.012). In conclusion, there is considerable overlap in common CT findings of MAC and M. abscessus pulmonary infection; however, lobar volume loss, nodule, airspace consolidation, and thin-walled cavity are more frequently seen in MAC than M. abscessus infection.

Nontuberculous mycobacterial (NTM) infections are an increasingly recognized cause of chronic lung disease in immunocompetent adults, and the M. avium complex, M. kansasii, and a rapidly growing mycobacteria such as M. abscessus, M. fortuitum, and M. chelonae account for most of the pathogens involved. Because the clinical features of NTM disease are not distinguishable from those of tuberculosis, and NTM are ubiquitous in the environment, diagnosis requires that the bacilli are isolated and identified. NTM diseases have been difficult to treat, though since the introduction of new macrolides, the outcome for patients with some NTM diseases has improved significantly. For correct diagnosis and the successful treatment of NTM pulmonary disease, a knowledge of the full spectrum of clinical and radiological findings is important.

OBJECTIVE: to assess the reliability of the inclination angle and anteversion of acetabular cup component in patients with idiopatic osteoarthritis of the hip, aseptic necrosis and hip neck fracture using trigonometric formula and plain radiographs. METHODS: 66 patients underwent cemented total arthroplasty of 72 hips. The inclination of acetabular component was measured using plain radiograph. The acetabular component anteversion was measured using trigonometric formula. RESULTS: it was observed that, in the osteoarthritic hips, hip neck fracture and aseptic necrosis, the degree of agreement was highly significant (p < 0.0001), in the measurements of anteversion and inclination angles, among the three assessments, from intra as well as inter-observers. All the agreement pairs were of excellent degree (ICC > 0.80). CONCLUSION: using plain radiographs and trigonometric formula, the method resulted to be highly accurate and reliable. Besides being easy to be calculated. No significant variation was found in the anteversion and inclination angles when compared with osteoarthritis of the hip, aseptic necrosis and hip neck fracture. OBJETIVO: Medir a confiabilidade do ângulo de inclinação e a anteversão do componente acetabular em pacientes com osteoartrose idiopática do quadril, necrose asséptica e na fratura do colo do fêmur por meio de uma fórmula trigonométrica e radiografias convencionais. MÉTODOS: Foram tratados 66 pacientes com artroplastia total cimentada em 72 quadris. A inclinação do componente acetabular foi medida por radiografias panorâmicas de bacia em incidência anteroposterior. A anteversão do componente acetabular foi medida com o uso de fórmula trigonométrica. RESULTADOS: Observou-se que, tanto nos quadris com artrose, na fratura do colo do fêmur e na necrose asséptica, o grau de concordância foi altamente significativo (p < 0,0001), nas medidas dos ângulos de anteversão e inclinação, entre os três avaliadores, tanto intra como interobservador. Todos os pares de concordância foram de grau ótimo (ICC > 0,80). CONCLUSÃO: Usando radiografias convencionais e uma fórmula trigonométrica, o método mostrou ser altamente preciso, fácil de ser calculado e com grande confiabilidade. Não foi encontrada variação significativa no ângulo de anteversão e no ângulo de inclinação quando comparado com a artrose do quadril, a necrose asséptica e a fratura do colo do fêmur.