A significant knowledge gap exists concerning the geographical distribution of nontuberculous mycobacteria (NTM) isolation worldwide.To provide a snapshot of NTM species distribution, global partners in the NTM-Network European Trials Group (NET) framework (www.ntm-net.org), a branch of the Tuberculosis Network European Trials Group (TB-NET), provided identification results of the total number of patients in 2008 in whom NTM were isolated from pulmonary samples. From these data, we visualised the relative distribution of the different NTM found per continent and per country.We received species identification data for 20 182 patients, from 62 laboratories in 30 countries across six continents. 91 different NTM species were isolated. Mycobacterium avium complex (MAC) bacteria predominated in most countries, followed by M. gordonae and M. xenopi. Important differences in geographical distribution of MAC species as well as M. xenopi, M. kansasii and rapid-growing mycobacteria were observed.This snapshot demonstrates that the species distribution among NTM isolates from pulmonary specimens in the year 2008 differed by continent and differed by country within these continents. These differences in species distribution may partly determine the frequency and manifestations of pulmonary NTM disease in each geographical location. @ERSpublications Species distribution among nontuberculous mycobacteria isolates from pulmonary specimens is geographically diverse

Objectives Information on the recently COVID‐19‐associated pulmonary aspergillosis (CAPA) entity is scarce. We describe eight CAPA patients, compare them to colonised ICU patients with coronavirus disease 2019 (COVID‐19), and review the published literature from Western countries. Methods Prospective study (March to May, 2020) that included all COVID‐19 patients admitted to a tertiary hospital. Modified AspICU and European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) criteria were used. Results COVID‐19‐associated pulmonary aspergillosis was diagnosed in eight patients (3.3% of 239 ICU patients), mostly affected non‐immunocompromised patients (75%) with severe acute respiratory distress syndrome (ARDS) receiving corticosteroids. Diagnosis was established after a median of 15 days under mechanical ventilation. Bronchoalveolar lavage was performed in two patients with positive Aspergillus fumigatus cultures and galactomannan (GM) index. Serum GM was positive in 4/8 (50%). Thoracic CT scan findings fulfilled EORTC/MSG criteria in one case. Isavuconazole was used in 4/8 cases. CAPA‐related mortality was 100% (8/8). Compared with colonised patients, CAPA subjects were administered tocilizumab more often (100% vs. 40%, p = .04), underwent longer courses of antibacterial therapy (13 vs. 5 days, p = .008), and had a higher all‐cause mortality (100% vs. 40%, p = .04). We reviewed 96 similar cases from recent publications: 59 probable CAPA (also putative according modified AspICU), 56 putative cases and 13 colonisations according AspICU algorithm; according EORTC/MSG six proven and two probable. Overall, mortality in the reviewed series was 56.3%. Conclusions COVID‐19‐associated pulmonary aspergillosis must be considered a serious and potentially life‐threatening complication in patients with severe COVID‐19 receiving immunosuppressive treatment.

We evaluated 117 isolates of Mycobacterium tuberculosis for susceptibility to linezolid by the proportion and E-test methods. Linezolid showed high in vitro activity, with all the strains inhibited by <1 g of the drug per ml. E-test MICs were at least 4 dilutions lower than their equivalents by the standard proportion method.Linezolid is an oxazolidinone whose mechanism of action involves inhibition of protein synthesis at a very early stage (7). This new drug has shown good activity against some grampositive bacteria, including resistant staphylococci, enterococci, and pneumococci (2, 5, 9). However, information in the literature regarding the activity of linezolid against Mycobacterium tuberculosis strains is scarce.We evaluated the in vitro activities of linezolid using the standard and E-test methods against 117 clinical isolates of M. tuberculosis with different levels of susceptibility to first-line antituberculous drugs.(This study was presented in part at the 41st Interscience Conference on Antimicrobial Agents and Chemotherapy, Chicago, Ill., 16 to 19 December 2001.) The active substance as reference powder and the linezolid E-test strips were provided by Pharmacia Upjohn (Barcelona, Spain). Stock solution was prepared at 10,000 g/ml in distilled water. Aliquots of the oxazolidinone were frozen at Ϫ70°C until use. Staphylococcus aureus strain ATCC 29213 was used as a quality control to assure the potency of the drug and the E-test activity.A total of 117 isolates of M. tuberculosis from different patients were selected from our laboratory collection (isolates collected in 1988 to 2000). Susceptibility testing on first-line antituberculous drugs (streptomycin, isoniazid, rifampin, and ethambutol [SIRE]) was performed by the agar proportion method at a reference laboratory. Of all the strains tested, 44 strains (38%) were resistant to at least one SIRE (R-SIRE), 12 strains were resistant to both isoniazid and rifampin, while the remainder were susceptible to the four SIRE drugs (S-SIRE).To prevent bias in the performance of the study, there was a single initial inoculum for each isolate for both the proportion and E-test methods. This inoculum was prepared in Middlebrook 7H9 and was adjusted to be equivalent to that of a McFarland no. 3 standard. Furthermore, both methods were performed in a double-blind manner.The agar proportion method was performed as recommended by the National Committee for Clinical Laboratory Standards (6). Briefly, linezolid was added to 7H10 agar medium (Difco) supplemented with OADC (oleic acid, albumin, dextrose, and catalase) (Becton Dickinson) at 50 to 56°C by doubling the dilutions to yield a final concentration of 0.125 to 4 g/ml. Five milliliters of each concentration of antimycobacterial agent-containing medium was dispensed into plastic quadrant petri dishes. As a growth control, one quadrant in each plate was filled with 7H10 agar medium with no drug. The inoculum of each isolate was prepared by diluting the initial inoculum in Middlebrook 7H9 until the absorbance was ...

Microscopy images must be acquired at the optimal focal plane for the objects of interest in a scene. Although manual focusing is a standard task for a trained observer, automatic systems often fail to properly find the focal plane under different microscope imaging modalities such as bright field microscopy or phase contrast microscopy. This article assesses several autofocus algorithms applied in the study of fluorescence‐labeled tuberculosis bacteria. The goal of this work was to find the optimal algorithm in order to build an automatic real‐time system for diagnosing sputum smear samples, where both accuracy and computational time are important. We analyzed 13 focusing methods, ranging from well‐known algorithms to the most recently proposed functions. We took into consideration criteria that are inherent to the autofocus function, such as accuracy, computational cost, and robustness to noise and to illumination changes. We also analyzed the additional benefit provided by preprocessing techniques based on morphological operators and image projection profiling. © 2012 International Society for Advancement of Cytometry

Microbiology laboratories where MALDI-TOF MS is available can benefit from its capacity to identify most clinically interesting non-tuberculous mycobacteria in a rapid, reliable, and inexpensive manner.