RESUMEN
A novel slowly growing, non-chromogenic species of the class Actinobacteria was isolated from a human respiratory sample in Nebraska, USA, in 2012. Analysis of the internal transcribed spacer sequence supported placement into the genus Mycobacterium with high sequence similarity to a previously undescribed strain isolated from a patient respiratory sample from Oregon, USA, held in a collection in Colorado, USA, in 2000. The two isolates were subjected to phenotypic testing and whole genome sequencing and found to be indistinguishable. The bacteria were acid-fast stain-positive, rod-shaped and exhibited growth after 7-10 days on solid media at temperatures ranging from 25 to 42°C. Colonies were non-pigmented, rough and slightly raised. Analyses of matrix-assisted laser desorption ionization time-of-flight profiles showed no matches against a reference library of 130 mycobacterial species. Full-length 16S rRNA gene sequences were identical for the two isolates, the average nucleotide identity (ANI) between their genomes was 99.7â% and phylogenetic comparisons classified the novel mycobacteria as the basal most species in the slowly growing Mycobacterium clade. Mycobacterium avium is the most closely related species based on rpoB gene sequence similarity (92â%), but the ANI between the genomes was 81.5â%, below the suggested cut-off for differentiating two species (95â%). Mycolic acid profiles were more similar to M. avium than to Mycobacterium simiae or Mycobacterium abscessus. The phenotypic and genomic data support the conclusion that the two related isolates represent a novel Mycobacterium species for which the name Mycobacterium talmoniae sp. nov. is proposed. The type strain is NE-TNMC-100812T (=ATCC BAA-2683T=DSM 46873T).
Asunto(s)
Mycobacterium/clasificación , Filogenia , Sistema Respiratorio/microbiología , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Genes Bacterianos , Humanos , Mycobacterium/genética , Mycobacterium/aislamiento & purificación , Infecciones por Mycobacterium/microbiología , Ácidos Micólicos/química , Oregon , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
Treatment of non-tuberculous mycobacterial lung disease (NTM-LD) is challenging for several reasons including the relative resistance of NTM to currently available drugs and the difficulty in tolerating prolonged treatment with multiple drugs. Yet-to-be-done, large, multicenter, prospective randomized studies to establish the best regimens will also be arduous because multiple NTM species are known to cause human lung disease, differences in virulence and response to treatment between different species and strains within a species will make randomization more difficult, the need to distinguish relapse from a new infection, and the difficulty in adhering to the prescribed treatment due to intolerance, toxicity, and/or drug-drug interactions, often necessitating modification of therapeutic regimens. Furthermore, the out-of-state resident status of many patients seen at the relatively few centers that care for large number of NTM-LD patients pose logistical issues in monitoring response to treatment. Thus, current treatment regimens for NTM-LD is largely based on small case series, retrospective analyses, and guidelines based on expert opinions. It has been nearly 10 years since the publication of a consensus guideline for the treatment of NTM-LD. This review is a summary of the available evidence on the treatment of the major NTM-LD until more definitive studies and guidelines become available.
RESUMEN
Mycobacterium chelonae is a rapidly growing opportunistic nontuberculous mycobacterial (NTM) species that causes infections in humans and other hosts. Here, we report the draft genome sequence of Mycobacterium chelonae type strain ATCC 35752, consisting of 4.89 Mbp, 63.96% G+C content, 4,489 protein-coding genes, 48 tRNAs, and 3 rRNA genes.
RESUMEN
In drug development, there are typically a series of preclinical studies that must be completed with new compounds or regimens before use in humans. A sequence of in vitro assays followed by in vivo testing in validated animal models to assess the activity against Mycobacterium tuberculosis, pharmacology and toxicity is generally used for advancing compounds against tuberculosis in a preclinical stage. A plethora of different assay systems and conditions are used to study the effect of drug candidates on the growth of M. tuberculosis, making it difficult to compare data from one laboratory to another. The Bill and Melinda Gates Foundation recognized the scientific gap to delineate the spectrum of variables in experimental protocols, identify which of these are biologically significant, and converge towards a rationally derived standard set of optimized assays for evaluating compounds. The goals of this document are to recommend protocols and hence accelerate the process of TB drug discovery and testing. Data gathered from preclinical in vitro and in vivo assays during personal visits to laboratories and an electronic survey of methodologies sent to investigators is reported. Comments, opinions, experiences as well as final recommendations from those currently engaged in such preclinical studies for TB drug testing are being presented. Certain in vitro assays and mouse efficacy models were re-evaluated in the laboratory as head-to-head experiments and a summary is provided on the results obtained. It is our hope that this information will be a valuable resource for investigators in the field to move forward in an efficient way and that key variables of assays are included to ensure accuracy of results which can then be used for designing human clinical trials. This document then concludes with remaining questions and critical gaps that are in need of further validation and experimentation.
Asunto(s)
Antituberculosos/farmacología , Proteínas Bacterianas/efectos de los fármacos , Evaluación Preclínica de Medicamentos/métodos , Mycobacterium tuberculosis/efectos de los fármacos , Tuberculosis/tratamiento farmacológico , Animales , Antituberculosos/farmacocinética , Modelos Animales de Enfermedad , Diseño de Fármacos , Evaluación Preclínica de Medicamentos/tendencias , Humanos , Ratones , Mycobacterium tuberculosis/patogenicidad , Tuberculosis/prevención & controlRESUMEN
Here we describe an experimental murine model that allows for aerosolized antituberculosis drug efficacy testing. Intrapulmonary aerosol delivery of isoniazid, capreomycin, and amikacin to mice with pulmonary infection of Mycobacterium tuberculosis demonstrated efficacy in reducing pulmonary bacterial loads similar to that seen by standard drug delivery methods, even when lower concentrations of drugs and fewer doses were used in the aerosolized drug regimens. Interestingly, intrapulmonary delivery of isoniazid also reduced the bacterial load in the spleen.
Asunto(s)
Antituberculosos/administración & dosificación , Antituberculosos/uso terapéutico , Isoniazida/administración & dosificación , Isoniazida/uso terapéutico , Pulmón/metabolismo , Tuberculosis Pulmonar/tratamiento farmacológico , Administración por Inhalación , Animales , Femenino , Pulmón/microbiología , Ratones , Ratones Endogámicos BALB C , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/patogenicidadRESUMEN
Azithromycin is a potent macrolide antibiotic with poorly understood antiinflammatory properties. Long-term use of azithromycin in patients with chronic inflammatory lung diseases, such as cystic fibrosis (CF), results in improved outcomes. Paradoxically, a recent study reported that azithromycin use in patients with CF is associated with increased infection with nontuberculous mycobacteria (NTM). Here, we confirm that long-term azithromycin use by adults with CF is associated with the development of infection with NTM, particularly the multi-drug-resistant species Mycobacterium abscessus, and identify an underlying mechanism. We found that in primary human macrophages, concentrations of azithromycin achieved during therapeutic dosing blocked autophagosome clearance by preventing lysosomal acidification, thereby impairing autophagic and phagosomal degradation. As a consequence, azithromycin treatment inhibited intracellular killing of mycobacteria within macrophages and resulted in chronic infection with NTM in mice. Our findings emphasize the essential role for autophagy in the host response to infection with NTM, reveal why chronic use of azithromycin may predispose to mycobacterial disease, and highlight the dangers of inadvertent pharmacological blockade of autophagy in patients at risk of infection with drug-resistant pathogens.
Asunto(s)
Antibacterianos , Autofagia/efectos de los fármacos , Azitromicina , Fibrosis Quística/complicaciones , Fibrosis Quística/tratamiento farmacológico , Infecciones por Mycobacterium/etiología , Adulto , Animales , Antibacterianos/efectos adversos , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Azitromicina/efectos adversos , Azitromicina/farmacología , Azitromicina/uso terapéutico , Células COS , Chlorocebus aethiops , Fibrosis Quística/microbiología , Farmacorresistencia Bacteriana , Inhibidores Enzimáticos/farmacología , Células HeLa , Humanos , Lisosomas/metabolismo , Macrólidos/farmacología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/microbiología , Ratones , Ratones Endogámicos C57BL , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/metabolismo , Mycobacterium/efectos de los fármacos , Mycobacterium/patogenicidad , Mycobacterium/fisiología , Infecciones por Mycobacterium/microbiología , Fagosomas/efectos de los fármacos , Fagosomas/metabolismo , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Sirolimus/farmacologíaRESUMEN
Female BALB/cAnNCrl (n = 170; age, 6 to 9 wk) mice were infected by intravenous inoculation of 5 × 10(6) cfu Mycobacterium tuberculosis strain Erdman (ATCC 35801). Between day 52 and 5 mo after infection, 10 of the 170 mice infected according to this protocol developed torticollis, including mice in treatment groups that received combination antibiotic therapy of rifampin-pyrazinamide or moxifloxacin-rifampin-pyrazinamide. Torticollis did not develop in mice receiving isoniazid- rifampin-pyrazinamide therapy, nor was it present in the cohort of aerogenically infected mice. Affected mice were euthanized, and complete necropsy evaluation was performed on 4 mice. Gross necropsy evaluation revealed typical tuberculosis lesions in lungs of infected mice. Histologic evaluation of tissues revealed granulomatous otitis media with intralesional acid-fast bacilli consistent with Mycobacterium tuberculosis. These cases represent an unusual finding specific to the intravenous mouse model of Mycobacterium tuberculosis and may represent a model of a similar condition in humans that is known as tuberculous otitis media.
Asunto(s)
Modelos Animales de Enfermedad , Ratones , Mycobacterium tuberculosis/fisiología , Otitis Media/etiología , Tortícolis/etiología , Tuberculosis/complicaciones , Tuberculosis/tratamiento farmacológico , Animales , Antituberculosos/administración & dosificación , Antituberculosos/efectos adversos , Autopsia/veterinaria , Compuestos Aza/administración & dosificación , Compuestos Aza/efectos adversos , Quimioterapia Combinada , Oído Medio/microbiología , Oído Medio/patología , Femenino , Fluoroquinolonas , Inyecciones Intravenosas , Isoniazida/administración & dosificación , Ratones Endogámicos BALB C , Moxifloxacino , Otitis Media/microbiología , Otitis Media/patología , Pirazinamida/administración & dosificación , Pirazinamida/efectos adversos , Quinolinas/administración & dosificación , Quinolinas/efectos adversos , Rifampin/administración & dosificación , Rifampin/efectos adversos , Tortícolis/microbiología , Tortícolis/patología , Tuberculosis/patologíaRESUMEN
The continuing global epidemic of tuberculosis, the increasing rate of multidrug resistant (MDR) tuberculosis and the more recent emergence of extensively drug resistant (XDR) tuberculosis are great causes for concern. A major international effort is currently underway to optimize current drug therapies and to discover new drugs that are active against these organisms. This effort has created a pipeline of new candidate drugs at various stages of preclinical and early clinical evaluations. Major challenges still exist, however, varying from the standardization and application of current animal models and their application to drug discovery and characterization to the fact that our knowledge about the basic biology of the MDR and XDR strains of Mycobacterium tuberculosis is minimal at best.
Asunto(s)
Antituberculosos/farmacología , Tuberculosis/tratamiento farmacológico , Animales , Antituberculosos/uso terapéutico , Modelos Animales de Enfermedad , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Cobayas , Humanos , Pulmón/efectos de los fármacos , Pulmón/patología , Ratones , Mycobacterium tuberculosis/efectos de los fármacos , Ratas , Tuberculosis/epidemiología , Tuberculosis/patología , Tuberculosis Resistente a Múltiples Medicamentos/tratamiento farmacológicoRESUMEN
We performed a retrospective cohort study of human immunodeficiency virus (HIV)-associated Mycobacterium avium complex (MAC) infection during 1997-2004 to assess for factors associated with initial macrolide resistance. Of 52 HIV-associated sterile-site MAC isolates, 9 (17%) were initially macrolide resistant. Prior MAC treatment and prolonged MAC prophylaxis with macrolides were associated with infection with a macrolide-resistant isolate; 2 (22%) of 9 patients had limited or no prior macrolide exposure. Initial MAC isolates in HIV-infected persons should routinely be tested for macrolide resistance.