Multiplex-PCR for differentiation of Mycobacterium bovis from Mycobacterium tuberculosis complex

We evaluated a multiplex-PCR to differentiate Mycobacterium bovis from M. tuberculosis Complex (MTC) by one step amplification based on simultaneous detection of pncA 169C > G change in M. bovis and the IS6110 present in MTC species. Our findings showed the proposed multiplex-PCR is a very useful tool for complementation in differentiating M. bovis from other cultured MTC species.

Antimycobacterial activity of some Brazilian indigenous medicinal drinks

Tuberculosis (TB) is a very serious problem worldwide and the increasing number of multiple drugs resistant TB cases makes the search for new anti-TB drugs an urgent need. Indigenous knowledge about the use of native plants to treat illnesses has contributed to the discovery of new medicines. In this study, the antimycobacterial activity ofseven medicinal drinks was assessed: Ananas sativus (hydroalcoholic fruit extract), Aristolochia triangularis(aqueous and hydroalcoholic leaf, root and stem extracts), Bromelia antiacantha (hydroalcoholic fruit extract), Stryphnodendron adstringens (hydroalcoholic bark extract), Tabebuia ovellanedae (hydroalcoholic bark extract), Vernonia polyanthes (hydroalcoholic root extract), all used by the Vanuíre indigenous community in the treatment of respiratory diseases. The activity was evaluated by using a time-to-kill assay, in which Mycobacterium tuberculosis H37Rv was cultured on Lowenstein-Jensen medium, after thirty minutes, one, three, six, twelve and twenty-four hours contact of the bacteria with each drink. Within half to one hour contact, the hydroalcoholic drinks of A. triangularis, S. adstringens, T. ovellanedae and V. polyanthes reduced the bacterial growth by 2 orders of magnitude in CFU/mL, and all bacterial growth was absent after three hours contact. In contrast, no mycobactericidal effect was detected in the aqueous extract of A. triangularis or in the hydroalcoholic beverages of A. sativus and B. antiacantha, even aftertwenty-four hours contact.

Rapid molecular diagnosis of tuberculosis and other mycobacteriosis in smear-negative clinical specimens

Delay in diagnosis of pulmonary and other forms of tuberculosis (TB) can be fatal, particularly in HIV-infected patients. Hence, techniques based on nucleic acid amplification, which are both rapid and of high specificity and sensitivity, are now widely used and recommended for laboratories that diagnose TB. In the present study, diagnostic methods based on mycobacterial DNA amplification were evaluated in comparative trials alongside tradicional bacterial methods, using negative smear samples from patients with clinically-suspected TB (sputum samples from 25 patients with suspected pulmonary TB, urine samples from two patients with suspected renal TB and cerebrospinal fluid samples from one patient with suspected meningeal TB). A specificity of 100% was achieved with DNA amplification methods and tradicional culture/identification methods, in relation to clinical findings and treatment results. For the smear-negative sputa, conventional PCR for M.tuberculosis was positive in 62% of suspected lung TB case, showing the same sensitivity as bacterial identification. Both techniques failed in the detection of extra-pulmonary samples. Nested PCR showed, after species-specific amplification, a sensitivity of 100% for M. avium and 85% for M. tuberculosis. For extra-pulmonary smear-negative samples, only Nested PCR detected M. tuberculosis and all cases were confirmed clinically. Nested PCR, in which two-step amplification reactions are performed, can identify the two most important mycobacteria in human pathology quickly and directly from clinical spicimens

Isolation of Mycobacterium gordonae from poultry slaughterhouse water in SÒo Paulo State, Brazil

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

) and V (33.3

Plantas do cerrado brasileiro com atividade contra Mycobacterium fortuitum / Brazilian cerrado plants active against Mycobacterium fortuitum

Mycobacterium fortuitum é uma micobactéria de crescimento rápido, ubíquo na natureza e relacionada a micobacteriose de importância médica.Ela tem sido isolada de bacteremias, abscessos, endocardites, feridas cirúrgicas e traumáticas.De difícil tratamento, o bacilo é reconhecido na literatura como resistente inclusive aos medicamentos utilizados na terapêutica da tuberculose.O objetivo deste trabalho foi pesquisar extratos vegetais do Cerrado brasileiro com atividade contra M. fortuitum, empregando a técnica do Microplate Alamar Blue Assay (MABA) como método analítico.Dos 26 extratos testados frente ao M.fortuitum, o extrato apolar de Quassia amara (extrato diclorometanico) foi o que apresentou melhor resultado com valor de CIM de 62,5mg/mL seguidos pelos extratos apolares de Syngonanthus macrolepsis, Davilla elliptica, Turnera ulmifolia com CIM de 125g/mL.Para as mesmas plantas analisadas, utilizando-se agentes extratores polares (etanol e metanol), foram verificados CIM superiores a 500g/mL.Os valores foram semelhantes aos de extratos de outras plantas analisadas sendo considerados não promissores.

Caracterização da população portadora de tuberculose do município de Américo Brasiliense/SP, no período de 1992 a 2002 / Caracterization of tuberculosis patients of Americo Brasilienses's city on 1992 to 2002

A tuberculose é um grave problema de Saúde Pública em todo o mundo, sendo desde 1993 considerada pela OMS uma emergência global. O Brasil ocupa a 13ª posição no ranking mundial em incidência de tuberculose. Ocorrências de tuberculose nas grandes cidades brasileiras são bem conhecidas e estudadas. Entretanto, pouco se sabe sobre a problemática da tuberculose nas pequenas cidades. Neste sentido, este trabalho visou obter as características epidemiológicas da população portadora de tuberculose do município de Américo Brasiliense, SP, no período de 1992 a 2002. Os resultados mostraram que a incidência de tuberculose nesta cidade teve picos coincidentes com os anos de intensa migração de mão de obra, sobretudo para o corte da cana de açúcar e que a tuberculose acomete principalmente os trabalhadores rurais do sexo masculino, na idade produtiva dos 20 aos 40 anos. Foi verificado também que a tuberculose pulmonar é a principal forma clínica e que os índices de abandono de tratamento e de cura, foram respectivamente de 1,8% e aproximadamente, de 90%. O índice de detecção da doença pela baciloscopia foi de cerca de 60%