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Concept: Acid fast bacilli


Mycobacteria are shaped by a thick envelope made of an array of uniquely structured lipids and polysaccharides. However, the spatial organizations of these molecules remain unclear. Here we show that exposure to an esterase from Mycobacterium smegmatis (Msmeg_1529), hydrolyzing the ester linkage of trehalose dimycolate (TDM) in vitro, triggers rapid and efficient lysis of Mycobacterium tuberculosis, Mycobacterium bovis BCG, and Mycobacterium marinum. Exposure to the esterase immediately releases free mycolic acids, while concomitantly depleting trehalose mycolates. Moreover, lysis could be competitively inhibited by an excess of purified TDM and was abolished by a S124A mutation affecting the catalytic activity of the esterase. These findings are consistent with an indispensible structural role of trehalose mycolates in architectural design of the exposed surface of mycobacterial envelope. Importantly, we also demonstrate that the esterase-mediated rapid lysis of M. tuberculosis significantly improves its detection in paucibacillary samples.

Concepts: Tuberculosis, Mycobacterium, Mycobacterium tuberculosis, Mycobacterium smegmatis, Mycobacterium bovis, Corynebacterineae, Acid fast bacilli, Mycobacterium marinum


A 68-year-old man on peritoneal dialysis (PD) was hospitalized with the clinical picture of peritonitis. The patient was diagnosed with peritonitis caused by nontuberculous mycobacteria (NTM) according to positive Ziehl-Neelsen staining and negative Mycobacterium tuberculosis polymerase chain reaction results. Oral levofloxacin and clarithromycin, and later intraperitoneal imipenem were started. According to the anti-NTM susceptibility test results, oral minocycline was administered. The patient was treated for 6 months. He recovered without PD catheter removal; thus, PD was successfully continued. A genetic analysis identified the isolate as Mycobacterium iranicum. This is the first report of PD-related peritonitis caused by M. iranicum.

Concepts: DNA, Polymerase chain reaction, Tuberculosis, Mycobacterium, Mycobacterium tuberculosis, Mycobacterium avium complex, Ziehl-Neelsen stain, Acid fast bacilli


It has recently been shown that the anti-mycobacterial pro-drug thiacetazone (TAC) inhibits the conversion of double bonds of mycolic acid precursors into cyclopropyl rings in Mycobacterium bovis var BCG, M. marimum and M. chelonae by affecting the cyclopropyl mycolic acid synthases (CMASs) as judged by the build-up of unsaturated mycolate precursors. In our hands, TAC inhibits mycolic acid biosynthesis in Mycobacterium tuberculosis and M. kansasii with almost negligible accumulation of those precursors. Our observations that ‘de novo’ biosynthesis of all the mycolic acid families decreased upon TAC treatment prompted us to analyse the role of each one of the Type II Fatty Acid Synthase (FASII) enzymes. Overexpression of the hadABC operon, encoding the essential FASII dehydratase complex, but not of any of the remaining FASII genes acting on the elongation of fatty acyl chains leading to the synthesis of meromycolic acids, resulted in high level of resistance to TAC in M. tuberculosis. Spontaneous M. tuberculosis and M. kansasii TAC-resistant mutants isolated during this work revealed mutations in the hadABC genes strongly supporting our proposal that these enzymes are new players in the resistance to this anti-mycobacterial compound.

Concepts: Fatty acid, Tuberculosis, Mycobacterium, Mycobacterium tuberculosis, Fatty acid synthase, Corynebacterineae, Acid fast bacilli, Mycobacterium kansasii


SETTING: Tuberculous lymphadenitis is the most common form of lymphadenopathy; its main histopathological finding is granulomatous inflammation.OBJECTIVE: A reverse blot hybridisation assay, REBA Myco-ID(®), was applied to formalin-fixed paraffin-embedded (FFPE) tissue showing granulomatous lymphadenitis to define the causative agents.DESIGN: A total of 119 granulomatous lymphadenitis cases observed between 2000 and 2010 were studied. All tissue samples were treated by haematoxylin and eosin and Ziehl-Neelsen stain. Mycobacterium tuberculosis and non-tuberculous mycobacteria (NTM) were identified using the REBA Myco-ID assay, and resistance to rifampicin (RMP) and isoniazid (INH) was determined using REBA MTB-MDR(®).RESULTS: Of the 119 cases, 113 (95%) were positive with the REBA Myco-ID assay, while 20 (16.8%) were positive on acid-fast bacilli smear. Of the 113 positive REBA Myco-ID cases, 110 (92.43%) were identified as M. tuberculosis, 2 (1.7%) as NTM, and 1 (0.8%) as coinfection with M. tuberculosis and M. chelonae. Only 1 (0.9%) of the 110 M. tuberculosis cases was identified as RMP-resistant.CONCLUSION: REBA Myco-ID is a highly sensitive and specific assay for detecting M. tuberculosis and NTM. M. tuberculosis is the main cause of granulomatous lymphadenitis.

Concepts: Microbiology, Tuberculosis, Mycobacterium, Mycobacterium tuberculosis, Staining, Ziehl-Neelsen stain, Leprosy, Acid fast bacilli


Background: Extrapulmonary tuberculosis (EPTB) constitutes about 10% to 20% of all cases of tuberculosis in immunocompetent patients and more than 50% of the cases in HIV-positive individuals worldwide. Little information is available on the clonal diversity of Mycobacterium species in Ethiopia from EPTB. Methods: This study was carried out on smear-negative EPTB patients to molecularly characterize Mycobacterium tuberculosis complex strains. A questionnaire, smear staining, culture, deletion typing, and spoligotyping were employed. Results: The proportional distribution of EPTB and isolates did not vary substantially (p > 0.05) amongst the socio-demographic parameters considered in the current investigation. Out of 98 fine needle aspirates processed for culture, 36.7% (36/98) were positive for mycobacterial growth. Further speciation of those culture-positive isolates showed that 88.9% were M. tuberculosis and the remaining could be non-tuberculous mycobacterial species. Spoligotyping revealed 16 clusters out of which 2 were new to the SITVIT database. The most dominant spoligotypes were SIT54, SIT53, and SIT149 in decreasing order. SIT54, SIT134, SIT173, SIT345, SIT357, SIT926, SIT91088, and SIT1580 were reported for the first time in Ethiopia. The family with the highest frequency identified was M. tuberculosis family T1, followed by family 33. Most of the strains belonged to Euro-American (61.4%) and Indo-Oceanic (36.3%) lineages. Conclusions: The present study shows the importance of M. tuberculosis as a major cause of EPTB in the study area. Moreover, the majority of isolates of M. tuberculosis were found in clusters, suggesting the possibility of the existence of recent transmission. This warrants strengthening of the control programs for EPTB in the study area.

Concepts: Tuberculosis, Mycobacterium, Mycobacterium tuberculosis, Mycobacterium avium complex, Corynebacterineae, Acid fast bacilli, Mycobacterium africanum, Mycobacterium tuberculosis complex


A 49-year-old man who had recently emigrated from Myanmar presented with a 6-month history of rusty brown sputum with hemoptysis. A tuberculin skin test was positive, but sputum smears were negative for acid-fast bacilli, ova, and parasites.

Concepts: Bacteria, Parasites, Tuberculosis, Mantoux test, Tuberculin, Acid-fast, Acid fast bacilli, Rusty Brown


Mycobacterium bovis, one of several mycobacteria of the M. tuberculosis complex, is a global zoonotic pathogen that primarily infects cattle. Humans become infected by consuming unpasteurized dairy products from infected cows (1,2); possible person-to-person airborne transmission has also been reported (3). In April 2014, a man in Nebraska who was born in Mexico was determined to have extensive pulmonary tuberculosis (TB) caused by M. bovis after experiencing approximately 3 months of cough and fever. Four months later, a U.S.-born Hispanic girl from a nearby town who had been ill for 4-5 months was also determined to have pulmonary TB caused by M. bovis. The only social connection between the two patients was attendance at the same church, and no common dietary exposure was identified. Both patients had pulmonary cavities on radiography and acid-fast bacilli (AFB) on sputum-smear microscopy, indicators of being contagious (4). Whole-genome sequencing results of the isolates were nearly indistinguishable. Initial examination of 181 contacts determined that 39 (22%) had latent infection: 10 (42%) of 24 who had close exposure to either patient, 28 (28%) of 100 who were exposed to one or both patients in church, and one (2%) of 57 exposed to the second patient at a school. Latent infection was diagnosed in six contacts on follow-up examination, 2 months after an initial negative test result (4), for an overall latent infection rate of 25%. No infected contacts recalled consuming unpasteurized dairy products, and none had active TB disease at the initial or secondary examination. Persons who have M. bovis TB should be asked about consumption of unpasteurized dairy products (2), and contact investigations should follow the same guidance as for M. tuberculosis TB (4).

Concepts: Infectious disease, Microbiology, Tuberculosis, Mycobacterium, Mycobacterium tuberculosis, Cattle, Mycobacterium bovis, Acid fast bacilli


Mycobacteria are characterized by their impermeable outer membrane, which is rich in mycolic acids(1). To transport substrates across this complex cell envelope, mycobacteria rely on type VII (also known as ESX) secretion systems(2). In Mycobacterium tuberculosis, these ESX systems are essential for growth and full virulence and therefore represent an attractive target for anti-tuberculosis drugs(3). However, the molecular details underlying type VII secretion are largely unknown, due to a lack of structural information. Here, we report the molecular architecture of the ESX-5 membrane complex from Mycobacterium xenopi determined at 13 Å resolution by electron microscopy. The four core proteins of the ESX-5 complex (EccB5, EccC5, EccD5 and EccE5) assemble with equimolar stoichiometry into an oligomeric assembly that displays six-fold symmetry. This membrane-associated complex seems to be embedded exclusively in the inner membrane, which indicates that additional components are required to translocate substrates across the mycobacterial outer membrane. Furthermore, the extended cytosolic domains of the EccC ATPase, which interact with secretion effectors, are highly flexible, suggesting an as yet unseen mode of substrate interaction. Comparison of our results with known structures of other bacterial secretion systems demonstrates that the architecture of type VII secretion system is fundamentally different, suggesting an alternative secretion mechanism.

Concepts: Microbiology, Secretion, Structure, Tuberculosis, Mycobacterium, Mycobacterium tuberculosis, Corynebacterineae, Acid fast bacilli


We report a case of disseminated Mycobacterium genavense infection resulting from neutralizing anti-interferon-γ autoantibodies in the patient. We identified M. genavense targeting the hsp65 gene in an aspiration specimen of the lymph node. Adult-onset immunodeficiency caused by neutralizing anti-interferon-γ autoantibodies, in addition to HIV infection, can lead to disseminated nontuberculous mycobacterial infection.

Concepts: HIV, Immune system, Patient, Tuberculosis, Mycobacterium, Corynebacterineae, Acid fast bacilli, Mycobacterium genavense


Postmortem examination of a wild Asian elephant at Rajiv Gandhi National Park, India, revealed nodular lesions, granulomas with central caseation, and acid-fast bacilli in the lungs. PCR and nucleotide sequencing confirmed the presence of Mycobacterium tuberculosis. This study indicates that wild elephants can harbor M. tuberculosis that can become fatal.

Concepts: Bacteria, Tuberculosis, Mycobacterium, Mycobacterium tuberculosis, Ziehl-Neelsen stain, Elephant, Acid fast bacilli, Auramine-rhodamine stain