Transmission and prevalence of drug-resistant tuberculosis in a Brazilian setting under a directly observed therapy short-course strategy.

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

Transmission and prevalence of drug-resistant tuberculosis in a Brazilian setting under a directly observed therapy short-course strategy

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

Population structure and circulating genotypes of drug-sensitive and drug-resistant Mycobacterium tuberculosis clinical isolates in São Paulo state, Brazil.

São Paulo is the most populous Brazilian state and reports the largest number of tuberculosis cases in the country annually (over 18,500). This study included 193 isolates obtained during the 2nd Nationwide Survey on Mycobacterium tuberculosis Drug Resistance that was conducted in São Paulo state and 547 isolates from a laboratory based study of drug resistance that were analyzed by the Mycobacteria Reference Laboratory at the Institute Adolfo Lutz. Both studies were conducted from 2006 to 2008 and sought to determine the genetic diversity and pattern of drug resistance of M. tuberculosis isolates (MTC) circulating in São Paulo. The patterns obtained from the spoligotyping analysis demonstrated that 51/740 (6.9%) of the isolates corresponded to orphan patterns and that 689 (93.1%) of the isolates distributed into 144 shared types, including 119 that matched a preexisting shared type in the SITVIT2 database and 25 that were new isolates. A total of 77/144 patterns corresponded to unique isolates, while the remaining 67 corresponded to clustered patterns (n=612 isolates clustered into groups of 2-84 isolates each). The evolutionarily ancient PGG1 lineages (Beijing, CAS1-DEL, EAI3-IND, and PINI2) were rarely detected in São Paulo and comprised only 13/740, or 1.76%, of the total isolates; all of the remaining 727/740, or 98.24%, of the MTC isolates from São Paulo state were from the recent PGG2/3 evolutionary isolates belonging to the LAM, T, S, X, and Haarlem lineages, i.e., the Euro-American group. This study provides the first overview of circulating genotypes of M. tuberculosis in São Paulo state and demonstrates that the clustered shared types containing seven or more M. tuberculosis isolates that are spread in São Paulo state included both resistant and susceptible isolates.

Estudo bibliométrico da produção científica da atividade antifúngica dos compostos químicos gamma-terpineno, terpinene-4-ol e sabineno / Bibliometric study of the scientific production of the antifungal activity of the chemical compounds gamma-terpinene, terpinene-4-ol and sabinene

Este trabalho teve como objetivo conhecer a produção científica sobre a atividade antifúngica dos compostos químicos gamma-terpineno, terpinene-4-ol e sabineno, de óleos essenciais. Trata-se de um estudo descritivo, exploratório de abordagem quantitativa, realizado por meio de revisão da literatura, onde foi possível mapear a produtividade científica dos compostos bioativos citados. A busca foi realizada em fevereiro de 2021, abordando o período de 1964 a 2020, utilizando a fonte de informação PubMed/MEDLINE. Na pesquisa livre os termos gamma-terpinene and antifungal, terpinene-4-ol and antifungal e sabinene and antifungal e no MeSH Database - Medical Subject Heading Terms (Mesh Terms ­ MeSH) desenvolvido pela U.S National Library of Medicine, utilizando os termos indexados gamma-terpinene and antifungal agents, terpinene-4-ol and antifungal agentes e sabinene and antifungal agents. Foram selecionados e analisados após os critérios de inclusão 192 artigos científicos. A análise foi realizada de forma descritiva com apresentação de frequências absolutas e relativas. O número de citações por artigo foi realizado no Google Acadêmico. Por meio do VOSViewer® foi analisado a rede de colaboração por coautoria e por coocorrência dos termos de pesquisa. A Banaras Hindu University, Universidade Estadual Paulista (UNESP), Universidade Federal do Ceará, Universidade de Coimbra...(AU)

This work aimed to know the scientific production on the antifungal activity of the chemical compounds gamma-terpinene, terpinene-4-ol and sabinene, in essential oils. This is a descriptive, exploratory study with a quantitative approach, carried out through a literature review, where it was possible to map the scientific productivity of the aforementioned bioactive compounds. The search was carried out in February 2021, covering the period from 1964 to 2020, using the information source PubMed/MEDLINE. In the free search the terms gamma-terpinene and antifungal, terpinene-4-ol and antifungal and sabinene and antifungal and in the MeSH Database - Medical Subject Heading Terms (Mesh Terms) developed by the US National Library of Medicine, using the indexed terms gamma -terpinene and antifungal agents, terpinene-4-ol and antifungal agents and sabinene and antifungal agents. 192 scientific articles were selected and analyzed after the inclusion criteria. The analysis was performed descriptively with the presentation of absolute and relative frequencies. The number of citations per article was performed on Google Scholar. Through VOSViewer®, the collaboration network was analyzed by co-authorship and by cooccurrence of the search terms. Banaras Hindu University, State University of São Paulo (UNESP), Federal University of Ceará, University of Coimbra and University of Belgrade, Serbia are the institutions that published the most in the period. Most authors developed the research in public institutions. Iran, India and Brazil were the countries that published the most. The most published journals are located in the United Kingdom and the United States. The most studied family of medicinal plants were Lamiaceae, Apiaceae and Myrtaceae. The most researched medicinal plants in terms of antifungal activity were: Origanum vulgare L. (Oregano), Thymus vulgaris L. (Thyme), Rosmarinus officinalis L. (rosemary), Melaleuca alternifolia L. (Melaleuca), Origanum majorana L. (Marjoram). Candida albicans was the most studied species, followed by Aspergillus niger and Aspergillus flavus. The average life in years of selected articles in PubMed/MEDLINE - free search on gamma-terpinene was 6.5 years; 5-year terpinene-4-ol and 7-year sabinene and in the MeSH Database survey, 7-year gamma-terpinene, 5-year terpinene-4-ol and 8-year sabinene. The analysis of the correlation network by co-authorship and by co-occurrence of the terms used in the research of the selected articles, gamma-terpinene identified three clusters, terpinene-4-ol two clusters and sabinene only one cluster. Based on the analysis of this research with data indexed in the PubMed/MEDLINE information source, it was possible to identify a growth in the studies of chemical compounds isolated from essential oils. (AU

Cord factor detection and macroscopic evaluation of mycobacterial colonies: an efficient combined screening test for the presumptive identification of Mycobacterium tuberculosis complex on solid media.

OBJECTIVE: The rapid differentiation between Mycobacterium tuberculosis and nontuberculous mycobacteria is fundamental for patients co-infected with tuberculosis and HIV. To that end, we use two methods in our laboratory: detection of cord factor and PCR-restriction enzyme analysis (PRA). The objective of this study was to evaluate the accuracy of a screening test on solid medium as a rapid method for the presumptive identification of M. tuberculosis complex, considering costs and turnover time. METHODS: A total of 152 strains were submitted to a combined screening test, consisting of the detection of cord factor under microscopy (Ziehl-Neelsen staining) and evaluation of the macroscopic aspect of colonies, as well as to PRA, which was used as the gold standard. Costs were estimated by calculating the price of all of the materials needed for each test. RESULTS: The overall accuracy of cord factor detection alone was 95.4% (95% CI: 90.7-98.1%), and that of the combined screening test was 99.3% (95% CI: 96.4-100%). Cord factor detection costs US$ 0.25, whereas the PRA costs US$ 7.00. Results from cord factor detection are ready in 2 days, whereas PRA requires 4 days to yield results. CONCLUSIONS: The presumptive identification of M. tuberculosis using the macroscopic evaluation of colonies combined with cord factor detection under microscopy is a simple, rapid and inexpensive test. We recommend the combined screening test to rapidly identify M. tuberculosis in resource-poor settings and in less well-equipped laboratories while awaiting a definite identification by molecular or biochemical methods.

Maintenance of Mycobacterium tuberculosis on glass beads.

The intent of this study was to estimate the shelf life of Mycobacterium tuberculosis strains, and to observe the loss of viability in some of them from year to year. From 2000 to 2004, 10,015 cultures of M. tuberculosis were preserved by freezing on glass beads at -70 degrees C. With the expectation that the loss of viability might be around 5-10%/yr of storage, 730 strains were analyzed in order to establish the prevalence of recovery within a 5% margin of error. This study shows that 94% of the strains preserved at -70 degrees C on glass beads could be recovered within 30 days. The recovery rates for drug-susceptible and drug-resistant strains showed no significant differences. The growth rates and the number of strains that showed abundant growth before the 30th day of incubation represent important features, since the subculture of a strain preserved for future use ought to quickly produce abundant growth in order to avoid misinterpretation of the tests. Our experience indicates that storage of M. tuberculosis on glass beads at -70 degrees C is a suitable procedure for an active culture collection in a public health laboratory like ours, where maintenance of M. tuberculosis cultures is a complementary activity and must be quick, practical, effective, and economical.

Transmissão e prevalência da resistência de Mycobacterium tuberculosis, após a implantação do DOTS, no Município de Guarulhos no período de 2007 a 2011 / Transmission and prevalence of resistance of Mycobacterium tuberculosis, after DOTS implementation in the Municipality of Guarulohs in the period 2007-2011

A tuberculose (TB) é uma doença infecto-contagiosa de distribuição universal e até metade do século XX era considerada a maior causa de morte por doenças infecciosas no mundo. O Tratamento Diretamente Observado (TDO) é um componente importante da Estratégia de Tratamento Diretamente Observado de curta duração (DOTS), pois visa a melhoria da adesão do paciente ao tratamento, redução do abandono, aumento da cura e prevenção à resistência a drogas. A cidade de Guarulhos (SP), Brasil, tem uma população estimada de 1.244.518 habitantes e está localizado a 17 km do município de São Paulo. Em 2012, o município apresentou uma taxa de incidência de TB de 31/100.000 habitantes. O TDO foi implementado em Guarulhos em 1998 e foi intensificado a partir de 2004, com o apoio da USAID (U.S. Agency for International Development). Este estudo teve como objetivo descrever as características epidemiológicas dos pacientes com TB pulmonar (TBP) em TDO e tratamento auto-administrado (TAA), a prevalência de resistência aos medicamentos de primeira linha e a transmissão de cepas resistentes entre pacientes com TBP residentes em Guarulhos. Foram incluídos no estudo, pacientes com TBP residentes e tratados em Guarulhos que tiveram diagnóstico confirmado por cultura, e que iniciaram tratamento no período de outubro de 2007 a outubro de 2011. Foram analisados retrospectivamente, dados demográficos, clínicos e laboratoriais obtidos no Instituto Adolfo Lutz, Centro de Vigilância Epidemiológica do Estado de São Paulo, CVE-TB-SP, e no Laboratório Municipal de Saúde Pública de Guarulhos (LMSPG). Os Isolados de Mycobacterium tuberculosis foram testados quanto à resistência às drogas de primeira linha pelo método MGIT960 (TS) e caracterizados por RFLP-IS6110. Para a análise de transmissão, foram incluídos os pacientes identificados no banco de perfis de RFLP-IS6110 do estado de São Paulo, que residiam em Guarulhos e trataram no município de São Paulo no mesmo período do estudo...

Tuberculosis (TB) is an infectious disease of worldwide distribution and up to half of the twentieth century was considered the leading cause of death from infectious diseases worldwide. Directly Observed Treatment (DOT) is an important component of Directly Observed Treatment, Short Course (DOTS). DOT strategy has been an effective tool to improve the adherence to the treatment, to reduce the dropout, to increase the cure and prevent the drug resistance. The city of Guarulhos (SP), Brazil, had an estimated population of 1.244.518 inhabitants and it is located 17 Km from the city of São Paulo. In 2012, Guarulhos had a TB incidence rate of 31/100,000 inhabitants. DOT was implemented in 1998 and was intensified in 2004, with support from USAID (U.S. Agency for International Development). The aim of this study was to describe the epidemiological characteristics of pulmonary TB (PTB) patients treated under DOT and self-administered treatment (SAT), the prevalence of resistance to first-line drugs, and the transmission of resistant strains among PTB patients residing in Guarulhos. PTB patients residents and treated in Guarulhos who were confirmed by culture and started treatment between October 2007 and October 2011 were included in the study. For the transmission analysis, patients who were residents but treated in Sao Paulo City, identified at the IS6110-RFLP database of São Paulo State, were also included. We retrospectively analyzed, data obtained at the Instituto Adolfo Lutz, TB Surveillance System of Sao Paulo State (CVE-TB-SP), and at the Guarulhos Municipal Public Health Laboratory (LMSPG). Mycobacterium tuberculosis isolates were subjected to drug susceptibility testing by MGIT 960 (DST) and typed by IS6110-RFLP. In the study period, 1.302 TBP patients were notified at the CVE-TB-SP. Of these, 408 patients had positive culture and were submitted to the DST. Two hundred and thirty-nine (58.6%) patients received DOT (DOT Group)...

Cord factor detection and macroscopic evaluation of mycobacterial colonies: an efficient combined screening test for the presumptive identification of Mycobacterium tuberculosis complex on solid media / Detecção do fator corda e avaliação do aspecto macroscópico das colônias de micobactérias: um eficiente teste de triagem combinado para a identificação presuntiva do complexo Mycobacterium tuberculosis em meios só

OBJECTIVE: The rapid differentiation between Mycobacterium tuberculosis and nontuberculous mycobacteria is fundamental for patients co-infected with tuberculosis and HIV. To that end, we use two methods in our laboratory: detection of cord factor and PCR-restriction enzyme analysis (PRA). The objective of this study was to evaluate the accuracy of a screening test on solid medium as a rapid method for the presumptive identification of M. tuberculosis complex, considering costs and turnover time. METHODS: A total of 152 strains were submitted to a combined screening test, consisting of the detection of cord factor under microscopy (Ziehl-Neelsen staining) and evaluation of the macroscopic aspect of colonies, as well as to PRA, which was used as the gold standard. Costs were estimated by calculating the price of all of the materials needed for each test. RESULTS: The overall accuracy of cord factor detection alone was 95.4 percent (95 percent CI: 90.7-98.1 percent), and that of the combined screening test was 99.3 percent (95 percent CI: 96.4-100 percent). Cord factor detection costs US$ 0.25, whereas the PRA costs US$ 7.00. Results from cord factor detection are ready in 2 days, whereas PRA requires 4 days to yield results. CONCLUSIONS: The presumptive identification of M. tuberculosis using the macroscopic evaluation of colonies combined with cord factor detection under microscopy is a simple, rapid and inexpensive test. We recommend the combined screening test to rapidly identify M. tuberculosis in resource-poor settings and in less well-equipped laboratories while awaiting a definite identification by molecular or biochemical methods.

OBJETIVO: A diferenciação rápida entre Mycobacterium tuberculosis e micobactérias não-tuberculosas é fundamental para os pacientes coinfectados com tuberculose e HIV. Para tanto, utilizamos duas metodologias em nosso laboratório: detecção do fator corda e PCR-restriction enzyme analysis (PRA). O objetivo do estudo foi avaliar a acurácia desse teste de triagem em meio sólido como um método rápido para a identificação presuntiva do complexo M. tuberculosis, considerando custos e tempo de resultado. MÉTODOS: Foram processadas 152 cepas pelo teste de triagem combinado, que consistiu da detecção do fator corda por microscopia (esfregaço corado por Ziehl-Neelsen) e avaliação do aspecto macroscópico das colônias, e PRA (padrão ouro). Os custos foram estimados através da obtenção dos preços dos insumos necessários para a realização de cada teste. RESULTADOS: A acurácia da detecção do fator corda foi de 95,4 por cento (IC95 por cento: 90,7-98,1 por cento) e a do teste de triagem combinado foi de 99,3 por cento (IC95 por cento: 96,4-100 por cento). O custo da detecção do fator corda foi de R$ 0,60 e do PRA de R$ 16,00. Os resultados da detecção do fator corda estão prontos em 2 dias, ao passo que os de PRA necessitam de 4 dias. CONCLUSÕES: A identificação presuntiva de M. tuberculosis usando o aspecto macroscópico das colônias em conjunto com a detecção de fator corda por microscopia é um teste simples, rápido e de baixo custo. Recomendamos o teste de triagem combinado para rapidamente identificar M. tuberculosis em sítios com poucos recursos financeiros e em laboratórios menos equipados, enquanto se aguarda a identificação definitiva por métodos moleculares ou bioquímicos.

Influência da qualidade das culturas de Mycobacterium tuberculosis nos resultados negativos dos testes da pirazinamidase / Influence of Mycobacterium tuberculosis cultures quality conditions on negative results on pyrazinamidase testing

A Organização Mundial da Saúde recomenda o uso do teste da pirazinamidase (PZAse) como método alternativo para determinação da resistência do Mycobacterium tuberculosis à pirazinamida, por ser um teste rápido e de fácil execução. Foram objetivos deste estudo: verificar a reprodutibilidade dos resultados negativos do teste da pirazinamidase quando realizado a partir das culturas originais e de seus subcultivos e relacioná-los com a qualidade das culturas originais e com os perfis de suscetibilidade à estreptomicina (S), isoniazida (I), rifampicina (R) e etambutol (E). Foram analisadas 115 culturas de Mycobacterium tuberculosis cujos cultivos originais apresentaram resultados negativos no teste da PZAse, o que representa resistência à pirazinamida. A qualidade das culturas foi avaliada, anotada e um segundo teste foi realizado a partir de subcultivos jovens e abundantes. A concordância entre os resultados do primeiro e do segundo teste foi de 72,2% e a qualidade das culturas mostrou correlação com os resultados (p< 0,001). O teste da pirazinamidase é útil quando utilizado juntamente com técnicas de detecção de suscetibilidade às drogas S,I,R,E, desde que seja realizado a partir de cultivos com boa qualidade, que permitam a utilização de inóculo abundante.

The pyrazinamidase is a fast and easy to perform assay, recommended by the World Health Organization as an alternative technique to determine pyrazinamide resistant Mycobacterium tuberculosis strains. This study aimed to assess the reproducibility of negative results on pyrazinamidase assay using both primary cultures and respective subcultures, and to correlate them with original cultures quality, as well as their susceptibility profile to streptomycin (S), isoniazid (I), rifampin (R) and ethambutol (E). A total of 115 Mycobacterium tuberculosis cultures were analyzed, which the original growth produced negative results on pyrazinamidase assay, implying a resistance to pyrazinamide. The cultures quality was assessed and recorded; a second testing was performed using recent and abundant subcultures. Results from the first and the second tests demonstrated an agreement rate of 72.2%, and the cultures quality showed correlation with the results (p<0.001). Pyrazinamidase testing is useful when it is combined with other techniques for analyzing mycobacteria susceptibility to S, I, R, E since it is performed with high quality cultures which allow the use of abundant inocula.

Cabine de segurança biológica: efeito da luz ultravioleta nas micobactérias / Biological safety cabinet: utraviolet radiation effect on mycobacteria

A cabine de segurança biológica (CSB) é o principal equipamento para efetuar a contenção de aerossóis produzidos nos procedimentos laboratoriais e a descontaminação com lâmpada UV, 15 minutos antes do início das atividades e 15 minutos após utilização da cabine é parte das boas práticas de laboratório. O objetivo deste estudo foi avaliar a ação da lâmpada UV da CSB classe II B2, em diversas espécies de micobactérias e correlacionar com o tempo de exposição. Cepas de referência foram subcultivadas, semeadas e incubadas a 37ºC até produzir turvação compatível com o tubo 1 da escala de MacFarland. Foram semeados 100L de suspensão bacteriana em placas com meio 7H11; as placas foram cobertas parcialmente com papel alumínio e expostas à radiação UV durante 5 e 10 minutos. Após exposição, os papéis foram retirados e as placas incubadas a 37ºC por 30 dias. Todas as placas apresentaram inibição de crescimento de bactérias na porção da placa em que houve exposição direta à radiação UV. Os resultados obtidos mostraram que a prática de utilização da radiação UV por 15 minutos após o uso da cabine e antes de iniciar outra atividade técnica, garante descontaminação adequada da CSB. Esta prática de biossegurança é recomendável para descontaminar a própria CSB e os materiais que são retirados da cabine.