Diversity Of Nontuberculous Mycobacteria Species Research Articles

ВИДЫ НЕТУБЕРКУЛЕЗНЫХ МИКОБАКТЕРИЙ, ЦИРКУЛИРУЮЩИХ НА ТЕРРИТОРИИ СИБИРСКОГО ФЕДЕРАЛЬНОГО ОКРУГА

Introduction. Non-tuberculous mycobacteria are ubiquitous; they are widespread in soil and water, including plumbing, aquariums, hot water supply systems, and air conditioning systems. Currently, more than 200 species of non-tuberculous mycobacteria are known, of which about 60 species have proven clinical significance, the rest are widespread in the environment, and little is known about their ability to cause diseases. Aim. The purpose of this study was to investigate the species diversity of non-tuberculous mycobacteria circulating in the Siberian Federal District of Russia. Materials and Methods. Two typical regions of the Siberian Federal District were selected for the study: Novosibirsk and Tomsk regions. Mycobacteria were cultured from patients’ sputum samples: 45 samples from the State Regional Clinical Tuberculosis Hospital and 106 from the Regional Department of Tomsk Phthisiopulmonology Medical Center. Further, non-tuberculous mycobacteria were identified by species, using the time-of-flight mass-spectrometry and the extraction with formic acid on an instrument according to the study protocol. Statistical processing of the study results was carried out using Microsoft Excel 2016 computer programs for Windows. Extensive indicators (the percentage of patients with detected pathogen drug resistance to the total number of patients, whose material was tested) were also calculated, and the boundaries of 95% confidence intervals (95% CI) were defined for the proportion (Fisher’s method). Statistical calculation of the research results was carried out in Microsoft Excel 2016. Results and Discussion. Among the slow-growing non-tuberculous mycobacteria species, Mycobacterium Avium complex is most common. Mycobacterium simie complex, Mycobacterium lentiflavum, and Mycobacterium parascorfulaceum were found less frequently. There were also Mycobacterium terrae complex and Mycobacterium nonchromogenicum found. The slow-growing mycobacteria, currently not grouped into a complex, include Mycobacterium Mycobacterium szulgae and Mycobacterium gordonae. Mycobacterium abscessus / Mycobacterium chelonae complex dominated among the fast-growing mycobacteria species. Mycobacterium fortuitum complex was a less common complex. Conclusions. For the first time, diversity was assessed and the dominant strains of mycobacteria circulating in the Novosibirsk and Tomsk regions were identified, which gives an idea of the main target strains, at which the main preventive and sanitizing measures should be targeted

Pulmonary mycobacteriosis: clinicalepidemiological profile of patients, treatment management, clinical outcome, and its species in the state of Mato Grosso, Brazil (2013-2020)

This study aimed to describe the clinical-epidemiological profile of patients in the State of Mato Grosso, who were affected by non-tuberculous mycobacteria (NTM), treatment management, clinical outcomes, and the species most commonly isolated from clinical samples. Between 2013 and 2020, the study analyzed 37 medical records from the Health Pulmonology sector of the Reference Center for Medium and High Complexity and it searched the Laboratory Environment Manager (GAL) system of the Central Public Health Laboratory of Mato Grosso. Patients with mycobacteriosis were predominantly diagnosed with MAC complex (avium-intracellulare complex) (n= 23.6%), with an average age of 62 years old; 51.4% were male, Caucasian (62%), and predominantly immunocompetent (HIV-negative). Most patients with comorbidities were older adults, they had sequelae of previous tuberculosis (56.8%), they were smokers (62.2%), and they had a low body mass index (29.7%) due to weight loss. Other patients had bronchiectasis (81.1%) and cavitary lesions (48.6%), with a lower incidence of chronic obstructive pulmonary disease (COPD). The municipalities with the highest rates of nontuberculous mycobacteria were Cuiabá (14 cases), Várzea Grande (five cases), Sinop (two cases), Nova Mutum (two cases), and Rondonópolis (two cases). This study presented data from the region, with a clinical, epidemiological profile, diversity of NTM species, and treatment management according to the patient’s response to the specific treatment of the identified Mycobacterium and various outcomes. KEY WORDS: Non-tuberculous mycobacteria; MAC complex Mycobacterium; comorbidities; tuberculosis; mycobacteriosis.

ЭТИОЛОГИЧЕСКАЯ ВЕРИФИКАЦИЯ МИКОБАКТЕРИОЗА ЛЕГКИХ ПРИ ИССЛЕДОВАНИИ ОПЕРАЦИОННОГО МАТЕРИАЛА У ПАЦИЕНТОВ С ДИАГНОЗОМ А16.0 – РЕАЛЬНАЯ ПРАКТИКА

Objective: to evaluate effectiveness of etiological verification of pulmonary mycobacteriosis (PM) in patients with diagnosed “pulmonary TB without bacteriological or histological confirmation” (А16.0) as per ICD-10 and species diversity of nontuberculous mycobacteria (NTM) in resection samples from the studied cohort. Materials and methods.We analyzed test results from 28 patients with initially diagnosed “pulmonary TB without bacteriological or histological confirmation” (А16.0) as per ICD-10 and verified PM according to resection sample studies. The microbiological study (at least two sputum samples, one bronchobiopsy sample, and one resection sample) included luminescent microscopy (LUM), extraction of M. tuberculosis DNA or NTM DNA by polymerase chain reaction (PCR), culture growth on liquid media in the ВАСТЕС MGIT 960 system, identification of mycobacteria by molecular genetic methods: real-time PCR for M. tuberculosis DNA detection (Syntol, Russia) and DNA-strip technology (GenoType Mycobacterium CM/AS Hain Lifescience). Results. The analysis of referral diagnoses established that in more than half of patients with diagnosed “pulmonary TB without bacteriological or histological confirmation” (А16.0) changes in the lungs had been referred to tuberculoma (64.3%) or infiltrative TB (21.4%); 28.6% of patients had received TB treatment as per chemotherapy regimen no. 3, and 17.8% of patients – broad-spectrum antibiotics. TB treatment duration varied from 4 to 28 months. Atypical lung resections were performed in most cases (75%). NTM DNA detection in resection samples was the most informative method of etiological verification of PMB (100% of cases). NTM culture growth was observed in 17.9% of cases. Only slowly-growing NTM were represented in the studied cohort as etiologically significant. Conclusion. The diagnosis verification is based on data obtained by microbiological and/or histological, cytological studies. Studies of resection samples from TB suspects significantly improve diagnostic effectiveness.

Trends and Species Diversity of Non-tuberculous Mycobacteria Isolated From Respiratory Samples in Northern China, 2014-2021.

BackgroundPulmonary non-tuberculous mycobacteria (NTM) infection has become a public health concern in China and around the world. The objective of this study was to describe the longitudinal changes in the frequency and diversity of NTM in northern China.MethodsWe retrospectively analyzed data on mycobacterium species in Beijing Chest Hospital from January 2014 to December 2021. The isolates were identified to species level by targeted DNA sequencing.ResultsAfter excluding duplicates, 1,755 NTM strains were analyzed, which were from 27 provinces in China over 8 years. Among all mycobacteria, the proportion of NTM increased each year, from 4.24% in 2014 to 12.68% in 2021. Overall, 39 different NTM species were identified, including 23 slow growing mycobacteria (SGM) and 16 rapid growing mycobacteria (RGM). The most common species were M. intracellulare (51.62%), M. abscessus (22.22%), M. kansasii (8.32%), M. avium (7.75%) and M. fortuitum (2.05%). The number of NTM species identified also increased each year from 9 in 2014 to 26 in 2021. Most species showed stable isolation rates over the years; however, the proportion of M. avium increased from 3.85 to 10.42% during the study period. Besides, 81 non-mycobacteria strains, including Gordonia (21 isolates), Nocardia (19 isolates) and Tsukamurella (17 isolates), etc., were also discovered.ConclusionThe proportion of NTM and species diversity increased considerably in northern China from 2014 to 2021. M. intracellulare was the most common NTM isolated among respiratory specimens, followed by M. abscessus and M. kansasii. Rare NTM species and non-mycobacteria pathogens also need attention.

Occurrence and risk factors of nontuberculous mycobacteria in tuberculosis-suspected patients in the north of Iran.

Background and Objectives:Some Nontuberculous Mycobacteria (NTM) can occasionally infect the human population and cause infections having symptoms similar to tuberculosis (TB). This study tried to provide updated data about the frequency and diversity of NTM species.Materials and Methods:Suspicious samples of Mycobacterium tuberculosis (MTB) with both positive results in Ziehl-Neelsen (ZN) staining and Löwenstein-Jensen medium culturing were evaluated during January 2016 and December 2018 in Gorgan, Iran. After determination of MTB isolates by the growth rate, pigmentation status, the niacin test, and the insertion sequence 6110 (IS6110) PCR assay, other unknown isolates (presumably NTM) were detected by the 16S rDNA sequencing method and drawing the phylogenetic tree. Based on the patients’ demographic information, their risk factors were also assessed.Results:Among 226 culture-positive samples, obtained from 2994 individuals with suspected symptoms of TB, the analyses found 12 (5.3%) NTM and three Mycobacterium caprae isolates. Mycobacterium simiae (6/12) was the most prevalent NTM species. The average nucleotide similarity value was 98.2% ± 3.7. In comparison to patients with MTB (211 confirmed cases), other mycobacterium infections were more common in patients over 65 years old (Odd ratio (95% convenience interval): 2.96 (0.69 – 12.59), P = 0.14).Conclusion:Although the NTM species has a small portion in TB suspected patients, their prevalence has increased, mainly in elderly patients. Moreover, M. simiae was the most prevalent NTM species in our region. Therefore, identification of common species in each region is recommended and clinicians should pay more attention to them in each region.

Profile of Nontuberculous Mycobacteria in Patients Suspected of Tuberculosis and Drug-Resistant Tuberculosis.

Objective Infections due to nontuberculous mycobacteria (NTM) is increasing globally and may present as drug-resistant tuberculosis (DRTB). In India, data on NTM prevalence and species diversity is limited. Present study was conducted to detect the prevalence and profile of NTM among patients suspected of DRTB using paraffin slide culture (PSC)and mycobacteria growth indicator tube (MGIT) culture methods for isolation of NTM. Material and Method A total of 2,938 samples suspected of TB/DRTB were cultured on PSC and MGIT960. Species identification of mycobacterial isolate was done by sequencing of 16s ribosomal RNA gene. Result Among 2938 samples, 35 (1.19%) were found positive for NTM by PSC and 9 (0.30%) were found positive by MGIT. The diversity of NTM species was high (13 species). Out of 35 NTM isolates by PSC, maximum 34.29% (12) isolates were found to be Mycobacterium fortuitum , followed by 11.43% (4) Mycobacterium abscessus and Mycobacterium chelonae , and 42.85% (15) were other species viz. 8.57% (3) were Mycobacterium intracellulare and Mycobacterium kansasii , 5.71% (2) were Mycobacterium peregrinum , and 2.85% (1) were Mycobacterium flavescens, Mycobacterium farcinogenes, Mycobacterium moriokanese, Mycobacterium wolinskyi, Mycobacterium simiae, Mycobacterium goodii , and Mycobacterium terrae each. Coinfection of Mycobacterium tuberculosis (MTB) and NTM was found in 60% (21) samples. Conclusion Prevalence of NTM was low among multidrug resistant tuberculosis/TB suspected patients, similar to other studies done in India. PSC was found better than MGIT for the isolation of NTM, though poor separation of NTM and MTB on subculture may have led to false negativity in cases of coinfection. About 13 species were isolated; M. fortuitum was the most common of all. Since coinfection of NTM and TB can also occur, samples of patients suspected of NTM should be cultured on PSC even if positive for MTB.

Laboratório Central de Saúde Pública do Piauí: frequência de espécies de micobactérias e aspectos epidemiológicos, 2014-2015

Background and Objectives: Knowledge about species diversity of non-tuberculous mycobacteria (NTM) and the frequency of tuberculosis (TB) is an important issue in rural-urban regions such as Piauí (northeast of Brazil), of low incidence rate of TB , can help to improve diagnosis and prevention strategies. The aim of this study is to examine some epidemiological aspects and the frequency of Mycobacterium tuberculosis (Mtb) and NTM isolated at the central public health reference laboratory, Dr. Costa Alvarenga, Piauí (LACEN-PI). Methods: Data records of all mycobacterosis and tuberculosis cases from January 2014 to March 2015 were analyzed. Results : Of the 20% (142/706) positive growths, 70% (99) were Mtb and 10% NTM. The remainde was of inadequate clinical samples, not allowing the identification of even the suspected NTM. The most frequent clinical form was pulmonary with TB patients younger than those infected with NTM (p = 0.001), the majority living in Teresina (52%). NTMs identified were M. abscessus (36%), M. avium, M. intracellulare, Mycobacterium sp. (14% each) and M. asiaticum, M. szulgai, M. kansasii 7% (each). Mtb drug resistance (7.8%) and TB co-infection with the human immunodeficiency virus (HIV-TB) found to be high (49%, 19/39) . Conclusion: The frequencies of Mtb infection, drug resistance and HIV-TB co-infection are still underestimated and failures in the identification of NTM may decrease the actual frequency of these infections. Therefore, there is a need for improvements in TB control and in the diagnosis of NTMs in Piauí.

Epidemiology of Non-Tuberculous Mycobacteria isolated from clinical specimens in Madrid, Spain, from 2013 to 2017.

The epidemiology of non-tuberculous mycobacteria (NTM) in Spain is largely unknown because systematic reporting is not compulsory. The aim of our study was to describe the frequency and diversity of NTM species in our region and their distribution according to the source sample, gender, and age of the patients. We performed a multicenter study of all NTM isolated in 24 public hospitals in Madrid from 2013 to 2017. A total of 6.923 mycobacteria were isolated: 4535 (65.5%) NTM, and 2.388 (34.5%) Mycobacterium tuberculosis complex (MTB). Overall, 61 different NTM species were identified. The most frequently isolated species were Mycobacterium avium complex (47.7%), M. lentiflavum (12.2%), M. gordonae (9.2%), M. fortuitum (8.9%), and M. abscessus (3.9%). Whereas MTB cases were stable during the study period, the number of NTM isolates increased considerably from 930 isolates in 2013 to 1012 in 2017; a sharp increase occurred in the last year. The rise in NTM isolates was mostly due to M. lentiflavum, M. kansasii, and M. abscessus mainly isolated from respiratory specimens in patients older than 60. The increase in isolation rate of NTM in our region is consistent with the increasing rates reported worldwide in the last decades. The rise in NTM isolates was mainly attributed to M. lentiflavum but it also should be noted the increasing of species with high pathogenic potential such as M. kansasii and M. abscessus.

Prevalence and species diversity of nontuberculous mycobacteria in drinking water supply system of Bahía Blanca City, Argentina.

There is evidence that tap water is the vehicle through which nontuberculous mycobacteria (NTM) infect or colonize the human body. The objective of this study was to determine the presence and diversity of NTM in the water distribution system of Bahía Blanca city, Argentina (sites S2/S3) and in the dike that supplies water to it (S1). Culture-dependent method, biochemical tests, and molecular method (16S rRNA sequencing gene) were combined to detect and identify NTM. NTM were isolated in 51.6% (64/124) of all the samples analyzed. Mycobacterium gordonae was the most frequently isolated organism (15/64) in all samples analyzed, followed by Mycobacterium peregrinum and Mycobacterium frederiksbergense. Significant differences were found in the residual chlorine values between sampling S2 and S3. In both sites, maximum counts were recorded but they did not correlate with low chlorine values. A concentration higher than 500 colony-forming unit/L of NTM was never found, which can be attributed to the negative effect caused by decontamination methods being a point to consider for the recovery of NTM. In 46.9% (30/64) of samples, both methods coincided in the identification, and the obtained sequences presented ≥99% identity. Identification at the species level was achieved in 50% (32/64) of the isolates. Nearly 17.2% (11/64) of the isolates showed a similarity <99%. It should be taken into account that sequencing of the 16S rRNA gene and biochemical tests are useful for the identification of several species, but it is necessary to incorporate other genes (hsp 65 and rpo B) to obtain accurate identification.

Nontuberculous mycobacterial lung disease in a high tuberculosis incidence setting in Brazil.

ABSTRACTObjective:The incidence of lung disease caused by nontuberculous mycobacteria (NTM) has been increasing worldwide. In Brazil, there are few studies about nontuberculous mycobacterial lung disease (NTMLD), and its prevalence is yet to be known. Our objective was to determine the specific etiology of the disease in the state of Rio Grande do Sul, Brazil, as well as the frequency and diversity of NTM species in our sample of patients. Methods:This is a retrospective analysis of the medical records of patients diagnosed with NTMLD treated in a referral center located in the city of Porto Alegre, Brazil, between 2003 and 2013. Results:Our sample comprised 100 patients. The most prevalent NTM species were Mycobacterium avium complex (MAC), in 35% of the cases; M. kansasii, in 17%; and M. abscessus, in 12%. A total of 85 patients had received previous treatment for tuberculosis. Associated conditions included structural abnormalities in the lungs, such as bronchiectasis, in 23% of the cases; COPD, in 17%; and immunosuppressive conditions, such as AIDS, in 24%. Conclusions:MAC and M. kansasii were the most prevalent species involved in NTMLD in the state, similarly to what occurs in other regions of Brazil. Data on regional epidemiology of NTMLD, its specific etiology, and associated conditions are essential to establish appropriate treatment, since each species requires specific regimens. Most patients with NTMLD had received previous tuberculosis treatment, which might lead to development of resistance and late diagnosis.

Identification and Characterization of Non-Tuberculous Mycobacteria Isolated from Extrapulmonary Tuberculosis Suspects by Heat Shock Protein 65 by PCR-RFLP

The genus Mycobacteria contains more than 170 officially recognized species. In the last decades, the reports of nontuberculous mycobacteria (NTM) associated with extrapulmonary diseases are increasing. The aim of this study is to know the prevalence of NTM among extrapulmonary tuberculosis cases in a tertiary care hospital in south India. This study was conducted from Mar 2017 to December 2017. Diagnosis was performed by Ziehl-Neelsen staining followed by culture in BacT/ALERT and Lowenstein Jensen culture medium. We applied conventional biochemical test like growth on the LJ medium containing PNBA (para nitro benzoic acid) to differentiate M.tuberculosis and NTM and PCR-restriction fragment length polymorphism analysis (PRA) of the hsp65 gene for species identification. We identified 96 non-tuberculous infection cases from 1138 (AFB)-Positive tuberculosis suspects. The diversity of NTM species were Mycobacterium abscessus 17(45.9%) followed by Mycobacterium fortuitum 10(27%), Mycobacterium chelonae 8(21.6%) and Mycobacterium simiae 2(5.4%). Regular documentation and reporting of these NTMs from clinical settings along with their sensitivity profile is essential to be aware of the clinical spectrum of disease associated and preferred treatment options.

Prevalence and species spectrum of both pulmonary and extrapulmonary nontuberculous mycobacteria isolates at a tertiary care center

Objective/background: Nontuberculous mycobacteria (NTM) infection associated with pulmonary and extrapulmonary disease has been increasing globally. Despite an increase in incidence rate of NTM infection, its prevalence, species diversity, and circulation pattern in India is largely unknown. This study sought to investigate the overall burden and diversity of NTM among both pulmonary and extrapulmonary clinical isolates from a Northern Indian population. Methods: The study was conducted in the Department of Microbiology, from January 2013 to December 2015. A total of 4620 clinical samples were collected from patients suspected to have pulmonary and extrapulmonary tuberculosis. Preliminary diagnosis was performed using Ziehl–Neelsen staining followed by liquid culture in BacT/ALERT three-dimensional system. A total of 906 positive cultures obtained were differentiated as either NTM or Mycobacterium tuberculosis complex using a biochemical and MPT64 antigen test. Further identification of NTM species was confirmed with a line probe assay. Results: Out of 906 cultures isolates, 263 (29.0%) were confirmed as NTM and 643 (71.0%) were identified as Mycobacterium tuberculosis complex. A total of 79.4% of the NTM were recovered from pulmonary and 18.2% from extrapulmonary specimens. The diversity of NTM species was high (13 species) and predominated by Mycobacterium abscessus (31.3%) followed by Mycobacterium fortuitum (22%), Mycobacterium intracellulare (13.6%), Mycobacterium chelonae (9.1%), however, M. abscessus and M. fortuitum were the predominant species in both types of clinical isolates. Men (60.4%) and older patients aged greater than 55years were the predominated risk group for NTM infection. Conclusion: The high prevalence and species diversity of NTM suggests the need for immediate and accurate characterization of NTM for proper treatment and management of patients.

Species diversity of non-tuberculous mycobacteria isolated from humans, livestock and wildlife in the Serengeti ecosystem, Tanzania

BackgroundNon-tuberculous mycobacteria (NTM), which are ubiquitous micro-organisms occurring in humans, animals and the environment, sometimes receive public health and veterinary attention as opportunistic disease-causing agents. In Tanzania, there is limited information regarding the diversity of NTM species, particularly at the human-livestock-wildlife interface such as the Serengeti ecosystem, where potential for cross species infection or transmission may exist.MethodsMycobacterial DNA was extracted from cultured isolates obtained from sputum samples of 472 suspect TB patients and 606 tissues from wildlife species and indigenous cattle. Multiplex PCR was used to differentiate NTM from Mycobacterium tuberculosis complex (MTBC) members. NTM were further identified to species level by nucleotide sequencing of the 16S rRNA gene.ResultsA total of fifty five (55) NTM isolates representing 16 mycobacterial species and 5 isolates belonging to the MTBC were detected. Overall, Mycobacterium intracellulare which was isolated from human, cattle and wildlife, was the most frequently isolated species (20 isolates, 36.4%) followed by M. lentiflavum (11 isolates, 20%), M. fortuitum (4 isolates, 7.3%) and M. chelonae-abscessus group (3 isolates, 5.5%). In terms of hosts, 36 isolates were from cattle and 12 from humans, the balance being found in various wildlife species.ConclusionThis study reveals a diversity of NTM species in the Serengeti ecosystem, some of which have potential for causing disease in animals and humans. The isolation of NTM from tuberculosis-like lesions in the absence of MTBC calls for further research to elucidate their actual role in causing disease. We are also suggesting a one health approach in identifying risk factors for and possible transmission mechanisms of the NTM in the agro-pastoral communities in the Serengeti ecosystem.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-014-0616-y) contains supplementary material, which is available to authorized users.

Increase in nontuberculous mycobacteria isolated in Shanghai, China: results from a population-based study.

BackgroundIn China, the prevalence of nontuberculous mycobacteria (NTM) in isolates from mycobacterial culture-positive patients with pulmonary tuberculosis (TB) is largely unknown.MethodsWe used conventional biochemical and 16S rRNA gene sequencing to identify species of mycobacteria in specimens from patients suspected of having TB. Drug-susceptibility testing was performed on NTM isolates using the proportion method. We also determined the independent risk factors associated with infection with NTM compared with infection with Mycobacterium tuberculosis.ResultsThe overall rate of NTM isolated from mycobacterial culture-positive patients was 5.9% in this population, with a significantly increasing trend from 3.0% in 2008 to 8.5% in 2012 (P for trend <0.001). The organism most frequently identified was M. kansasii (45.0%), followed by M. intracellulare (20.8%) and M. chelonae/abscessus (14.9%). The overall proportion of isolates resistant to the four first-line anti-TB agents were 64.6% for isoniazid, 77.6% for streptomycin, 63.3% for rifampicin and 75.1% for ethambutol. The risk factors most often associated with NTM infection were older age (P for trend <0.001), being a resident of Shanghai (adjusted odds ratio [aOR], 1.48; 95% CI, 1.10–2.00), having been treated for tuberculosis (aOR, 1.64; 95% CI, 1.18–2.29), having a cavity on chest X-ray (aOR, 1.51; 95% CI, 1.16–1.96), and being sputum smear–negative (aOR, 1.59; 95% CI, 1.16–2.18).ConclusionsThe prevalence of NTM isolated in Shanghai increased between 2008 and 2012, thus clinicians should consider NTM as a possible cause of TB-like disease. Accurate species identification is imperative so that proper treatment can be administered for diseases caused by the diversity of NTM species.