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1.
Sci Rep ; 14(1): 11187, 2024 05 16.
Article En | MEDLINE | ID: mdl-38755267

Mycobacteroides (Mycobacterium) abscessus, which causes a variety of infectious diseases in humans, is becoming detected more frequently in clinical specimens as cases are spreading worldwide. Taxonomically, M. abscessus is composed of three subspecies of M. abscessus subsp. abscessus, M. abscessus subsp. bolletii, and M. abscessus subsp. massiliense, with different susceptibilities to macrolides. In order to identify rapidly these three subspecies, we determined useful biomarker proteins, including ribosomal protein L29, L30, and hemophore-related protein, for distinguishing the subspecies of M. abscessus using the matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) profiles. Thirty-three clinical strains of M. abscessus were correctly identified at the subspecies-level by the three biomarker protein peaks. This study ultimately demonstrates the potential of routine MALDI-MS-based laboratory methods for early identification and treatment for M. abscessus infections.


Bacterial Proteins , Mycobacterium abscessus , Ribosomal Proteins , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Ribosomal Proteins/metabolism , Ribosomal Proteins/analysis , Mycobacterium abscessus/metabolism , Bacterial Proteins/metabolism , Humans , Mycobacterium Infections, Nontuberculous/microbiology , Mycobacterium Infections, Nontuberculous/diagnosis , Biomarkers/analysis , Biomarkers/metabolism
2.
BMC Microbiol ; 24(1): 172, 2024 May 18.
Article En | MEDLINE | ID: mdl-38760693

BACKGROUND: We evaluated whether the sputum bacterial microbiome differs between nontuberculous mycobacteria pulmonary disease (NTM-PD) patients with stable disease not requiring antibiotic treatment and those requiring antibiotics. METHODS: We collected sputum samples from 21 clinically stable NTM-PD patients (stable group) and 14 NTM-PD patients needing antibiotic treatment (treatment group). We also obtained 13 follow-up samples from the stable group. We analyzed the 48 samples using 16S rRNA gene sequencing (V3-V4 region) and compared the groups. RESULTS: In the linear discriminant analysis effect size (LEfSe) analysis, the species Porphyromonas pasteri, Haemophilus parahaemolyticus, Prevotella nanceiensis, and Gemella haemolysans were significantly more prevalent in the sputum of the stable group compared to the treatment group. No taxa showed significant differences in alpha-/beta-diversity or LEfSe between the 21 baseline and 13 follow-up sputum samples in the stable group. In the stable group, the genus Bergeyella and species Prevotella oris were less common in patients who achieved spontaneous culture conversion (n = 9) compared to those with persistent NTM positivity (n = 12) (effect size 3.04, p = 0.039 for Bergeyella; effect size 3.64, p = 0.033 for P. oris). In the treatment group, H. parainfluenzae was more common in patients with treatment success (n = 7) than in treatment-refractory patients (n = 7) (effect size 4.74, p = 0.013). CONCLUSIONS: Our study identified distinct bacterial taxa in the sputum of NTM-PD patients based on disease status. These results suggest the presence of a microbial environment that helps maintain disease stability.


Microbiota , Mycobacterium Infections, Nontuberculous , RNA, Ribosomal, 16S , Sputum , Humans , Sputum/microbiology , Male , Female , Microbiota/genetics , Microbiota/drug effects , Aged , Mycobacterium Infections, Nontuberculous/microbiology , Mycobacterium Infections, Nontuberculous/drug therapy , RNA, Ribosomal, 16S/genetics , Middle Aged , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Nontuberculous Mycobacteria/isolation & purification , Nontuberculous Mycobacteria/genetics , Nontuberculous Mycobacteria/classification , Nontuberculous Mycobacteria/drug effects , DNA, Bacterial/genetics , Lung Diseases/microbiology , Lung Diseases/drug therapy
3.
J Korean Med Sci ; 39(20): e167, 2024 May 27.
Article En | MEDLINE | ID: mdl-38804011

BACKGROUND: Coinfections with multiple nontuberculous mycobacterial (NTM) species have not been widely studied. We aimed to evaluate the clinical characteristics and treatment outcomes in patients with NTM-pulmonary disease (PD) caused by coinfection with multiple NTM species. METHODS: We retrospectively reviewed patients with NTM-PD at a tertiary referral hospital in Korea between March 2012 and December 2018. Coinfection was defined as two or more species of NTM pathogens isolated from the same respiratory specimen or different specimens within three months. RESULTS: Among 1,009 patients with NTM-PD, 147 (14.6%) NTM coinfections were observed (average age 64.7 years, 69.4% women). NTM species were identified more frequently (median 6 vs. 3 times, P < 0.001) in the coinfection group than in the single species group, and follow-up duration was also longer in the coinfection group (median 44.9 vs. 27.1 months, P < 0.001). Mycobacterium avium complex (MAC) and M. abscessus and M. massiliense (MAB) were the dominant combinations (n = 71, 48.3%). For patients treated for over six months in the MAC plus MAB group (n = 31), sputum culture conversion and microbiological cure were achieved in 67.7% and 41.9% of patients, respectively. We divided the MAC plus MAB coinfection group into three subgroups according to the target mycobacteria; however, no statistical differences were found in the treatment outcomes. CONCLUSION: In NTM-PD cases, a significant number of multiple NTM species coinfections occurred. Proper identification of all cultured NTM species through follow-up is necessary to detect multispecies coinfections. Further research is needed to understand the nature of NTM-PD in such cases.


Coinfection , Lung Diseases , Mycobacterium Infections, Nontuberculous , Nontuberculous Mycobacteria , Humans , Female , Male , Middle Aged , Retrospective Studies , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium Infections, Nontuberculous/complications , Mycobacterium Infections, Nontuberculous/diagnosis , Mycobacterium Infections, Nontuberculous/microbiology , Aged , Coinfection/microbiology , Nontuberculous Mycobacteria/isolation & purification , Treatment Outcome , Lung Diseases/microbiology , Lung Diseases/complications , Mycobacterium avium Complex/isolation & purification , Anti-Bacterial Agents/therapeutic use , Republic of Korea
4.
Int J Mycobacteriol ; 13(1): 34-39, 2024 Jan 01.
Article En | MEDLINE | ID: mdl-38771277

BACKGROUND: In patients with cystic fibrosis (CF), representatives of the fast-growing Mycobacterium abscessus complex (MABSc) are often distinguished, but the culture of the material taken from such patients increases the growth time. We analyzed the terms of cultivation of MABSc representatives on dense nutrient media and also evaluated the productivity of a modified nutrient medium based on agar for the isolation of Burkholderia cepacia complex (BCC). METHODS: Sixty-four strains of MABSc isolated from patients with CF and suspected tuberculosis were analyzed. The material from the patients was cultured on a universal chromogenic medium, 5% blood agar, yolk-salt agar, selective medium for isolation of BCC, and Löwenstein-Jensen medium. The cultures were incubated for 5 days (37°C, aerobic conditions), after for 23 days (28°C, aerobic conditions). The productivity of the developed nutrient medium was evaluated by the number of cells that gave visible growth after culturing 0.1 mL of a bacterial suspension of 103 CFU/mL. RESULTS: 76.8% of the strains grew in a 2-week period, and 23.2% of the strains were obtained at a later date from 18 to 28 days (average: 21.23 days). The modified medium with a concentration of 240 mg of iron (III) polymaltose hydroxide proved to be the most optimal for the isolation of MABSc. CONCLUSION: When using a chromogenic medium for culture material from patients with CF, it is necessary to extend incubation up to 28 days to increase the probability of MABSc isolation. The modified BCC medium showed a good selectivity result but required further investigation.


Culture Media , Cystic Fibrosis , Mycobacterium Infections, Nontuberculous , Mycobacterium abscessus , Humans , Cystic Fibrosis/microbiology , Culture Media/chemistry , Mycobacterium abscessus/growth & development , Mycobacterium abscessus/isolation & purification , Mycobacterium Infections, Nontuberculous/microbiology , Time Factors , Bacteriological Techniques/methods , Burkholderia cepacia complex/isolation & purification , Burkholderia cepacia complex/growth & development
5.
Life Sci Alliance ; 7(7)2024 07.
Article En | MEDLINE | ID: mdl-38744470

Developing effective tuberculosis drugs is hindered by mycobacteria's intrinsic antibiotic resistance because of their impermeable cell envelope. Using benzothiazole compounds, we aimed to increase mycobacterial cell envelope permeability and weaken the defenses of Mycobacterium marinum, serving as a model for Mycobacterium tuberculosis Initial hit, BT-08, significantly boosted ethidium bromide uptake, indicating enhanced membrane permeability. It also demonstrated efficacy in the M. marinum-zebrafish embryo infection model and M. tuberculosis-infected macrophages. Notably, BT-08 synergized with established antibiotics, including vancomycin and rifampicin. Subsequent medicinal chemistry optimization led to BT-37, a non-toxic and more potent derivative, also enhancing ethidium bromide uptake and maintaining synergy with rifampicin in infected zebrafish embryos. Mutants of M. marinum resistant to BT-37 revealed that MMAR_0407 (Rv0164) is the molecular target and that this target plays a role in the observed synergy and permeability. This study introduces novel compounds targeting a new mycobacterial vulnerability and highlights their cooperative and synergistic interactions with existing antibiotics.


Benzothiazoles , Drug Synergism , Mycobacterium marinum , Zebrafish , Animals , Benzothiazoles/pharmacology , Mycobacterium marinum/drug effects , Antitubercular Agents/pharmacology , Microbial Sensitivity Tests , Mycobacterium tuberculosis/drug effects , Humans , Anti-Bacterial Agents/pharmacology , Cell Membrane Permeability/drug effects , Macrophages/drug effects , Macrophages/microbiology , Macrophages/metabolism , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium Infections, Nontuberculous/microbiology , Cell Membrane/metabolism , Cell Membrane/drug effects , Rifampin/pharmacology
7.
J Infect Dev Ctries ; 18(4): 651-654, 2024 Apr 30.
Article En | MEDLINE | ID: mdl-38728638

INTRODUCTION: Mycobacterium marinum infection rarely occurs and has atypical symptoms. It is challenging to distinguish disseminated M. marinum infection from multifocal dermatosis caused by other factors clinically. CASE PRESENTATION: Herein, we reported a 68-year-old male patient with Human Immunodeficiency Virus (HIV) who presented redness and swelling in his left hand after being stabbed by marine fish for over 2 months. Mycobacterium tuberculosis infection was considered according to biochemical and pathological examinations, while empirical anti-infection treatment was ineffective. RESULTS: The metagenomic next-generation sequencing (mNGS) detected a large amount of M. marinum sequences, and the patient was finally diagnosed with M. marinum infection. After one month of combination therapy with ethambutol, rifabutin, moxifloxacin, and linezolid, the swelling disappeared significantly. In this case, the successful application of mNGS in diagnosing and treating M. marinum infection has improved the understanding of the microbe both in the laboratory and clinically, especially in patients with HIV. CONCLUSIONS: For diseases with atypical symptoms or difficulty in determining the pathogens, mNGS is suggested in clinical procedures for rapid and accurate diagnosis and treatment.


HIV Infections , Mycobacterium Infections, Nontuberculous , Mycobacterium marinum , Humans , Male , Mycobacterium Infections, Nontuberculous/diagnosis , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium Infections, Nontuberculous/microbiology , Aged , Mycobacterium marinum/isolation & purification , Mycobacterium marinum/genetics , HIV Infections/complications , High-Throughput Nucleotide Sequencing , Metagenomics , Ethambutol/therapeutic use , Anti-Bacterial Agents/therapeutic use
8.
PLoS One ; 19(5): e0303516, 2024.
Article En | MEDLINE | ID: mdl-38728330

Increasingly prevalent, nontuberculous mycobacteria (NTM) infections affect approximately 20% of people with cystic fibrosis (CF). Previous studies of CF sputum identified lower levels of the host metabolite itaconate in those infected with NTM. Itaconate can inhibit the growth of M. tuberculosis (MTB) in vitro via the inhibition of the glyoxylate cycle enzyme (ICL), but its impact on NTM is unclear. To test itaconic acid's (IA) effect on NTM growth, laboratory and CF clinical strains of Mycobacterium abscessus and Mycobacterium avium were cultured in 7H9 minimal media supplemented with 1-10 mM of IA and short-chain fatty acids (SCFA). M. avium and M. abscessus grew when supplemented with SCFAs, whereas the addition of IA (≥ 10 mM) completely inhibited NTM growth. NTM supplemented with acetate or propionate and 5 mM IA displayed slower growth than NTM cultured with SCFA and ≤ 1 mM of IA. However, IA's inhibition of NTM was pH dependent; as similar and higher quantities (100 mM) of pH adjusted IA (pH 7) did not inhibit growth in vitro, while in an acidic minimal media (pH 6.1), 1 to 5 mM of non-pH adjusted IA inhibited growth. None of the examined isolates displayed the ability to utilize IA as a carbon source, and IA added to M. abscessus isocitrate lyase (ICL) decreased enzymatic activity. Lastly, the addition of cell-permeable 4-octyl itaconate (4-OI) to THP-1 cells enhanced NTM clearance, demonstrating a potential role for IA/itaconate in host defense against NTM infections.


Succinates , Succinates/pharmacology , Succinates/metabolism , Humans , Hydrogen-Ion Concentration , Nontuberculous Mycobacteria/drug effects , Nontuberculous Mycobacteria/growth & development , THP-1 Cells , Mycobacterium Infections, Nontuberculous/microbiology , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium avium/drug effects , Mycobacterium avium/growth & development , Mycobacterium abscessus/drug effects , Mycobacterium abscessus/growth & development , Mycobacterium abscessus/metabolism
9.
Diagn Microbiol Infect Dis ; 109(3): 116296, 2024 Jul.
Article En | MEDLINE | ID: mdl-38640607

Pulmonary infection due to Mycobacterium abscessus complex (MABC) usually occurs in children with underlying risk factors including cystic fibrosis (CF), chronic lung disease, and immunocompromised status, but rarely in immunocompetent children without underlying lung disease, especially in infants. We present a case of MABC pulmonary disease (MABC-PD) in an otherwise healthy 53-day-old male infant with one week of cough and respiratory distress. Computed tomography showed multiple masses across both lungs. Isolated mycobacteria from his bronchoalveolar lavage fluid were identified as MABC. We describe our complete evaluation, including immunodeficiency evaluation incorporating whole exome sequencing and our therapeutic process given complicated susceptibility pattern of the M. abscessus isolate, and review literature for MABC-PD in immunocompetent children. The infant was successfully treated through prolonged treatment with parenteral Amikacin, Cefoxitin, Linezolid, and Clarithromycin, combined with inhaled Amikacin.


Anti-Bacterial Agents , Mycobacterium Infections, Nontuberculous , Mycobacterium abscessus , Humans , Male , Mycobacterium abscessus/drug effects , Mycobacterium abscessus/isolation & purification , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium Infections, Nontuberculous/microbiology , Mycobacterium Infections, Nontuberculous/diagnosis , Anti-Bacterial Agents/therapeutic use , Infant , Bronchoalveolar Lavage Fluid/microbiology , Amikacin/therapeutic use , Pneumonia, Bacterial/drug therapy , Pneumonia, Bacterial/microbiology , Pneumonia, Bacterial/diagnosis , Treatment Outcome , Tomography, X-Ray Computed , Clarithromycin/therapeutic use , Linezolid/therapeutic use
10.
Zhonghua Jie He He Hu Xi Za Zhi ; 47(4): 352-356, 2024 Apr 12.
Article Zh | MEDLINE | ID: mdl-38599811

With the development of testing technology, the diagnosis of nontuberculous mycobacterium (NTM) lung disease has gradually increased in recent years. Because the clinical characteristics of NTM are not typical, and its imaging manifestations are diverse and nonspecific, missed diagnosis and misdiagnosis are common. Etiological investigation is necessary for diagnosis. Conventional etiological investigations are very limited for the diagnosis of NTM. We reported a case of NTM lung disease presenting with a mass and atelectasis with mediastinal and hilar lymph node enlargement that resembled malignant tumors. The literature on this condition was reviewed to improve the clinician's understanding and broaden clinical thinking.


Lymphadenopathy , Mycobacterium Infections, Nontuberculous , Pulmonary Atelectasis , Humans , Nontuberculous Mycobacteria , Mycobacterium Infections, Nontuberculous/microbiology , Pulmonary Atelectasis/pathology , Lymph Nodes/pathology
11.
PLoS One ; 19(4): e0301659, 2024.
Article En | MEDLINE | ID: mdl-38640113

Clinical prediction of nontuberculous mycobacteria lung disease (NTM-LD) progression remains challenging. We aimed to evaluate antigen-specific immunoprofiling utilizing flow cytometry (FC) of activation-induced markers (AIM) and IFN-γ enzyme-linked immune absorbent spot assay (ELISpot) accurately identifies patients with NTM-LD, and differentiate those with progressive from nonprogressive NTM-LD. A Prospective, single-center, and laboratory technician-blinded pilot study was conducted to evaluate the FC and ELISpot based immunoprofiling in patients with NTM-LD (n = 18) and controls (n = 22). Among 18 NTM-LD patients, 10 NTM-LD patients were classified into nonprogressive, and 8 as progressive NTM-LD based on clinical and radiological features. Peripheral blood mononuclear cells were collected from patients with NTM-LD and control subjects with negative QuantiFERON results. After stimulation with purified protein derivative (PPD), mycobacteria-specific peptide pools (MTB300, RD1-peptides), and control antigens, we performed IFN-γ ELISpot and FC AIM assays to access their diagnostic accuracies by receiver operating curve (ROC) analysis across study groups. Patients with NTM-LD had significantly higher percentage of CD4+/CD8+ T-cells co-expressing CD25+CD134+ in response to PPD stimulation, differentiating between NTM-LD and controls. Among patients with NTM-LD, there was a significant difference in CD25+CD134+ co-expression in MTB300-stimulated CD8+ T-cells (p <0.05; AUC-ROC = 0.831; Sensitivity = 75% [95% CI: 34.9-96.8]; Specificity = 90% [95% CI: 55.5-99.7]) between progressors and nonprogressors. Significant differences in the ratios of antigen-specific IFN-γ ELISpot responses were also seen for RD1-nil/PPD-nil and RD1-nil/anti-CD3-nil between patients with nonprogressive vs. progressive NTM-LD. Our results suggest that multiparameter immunoprofiling can accurately identify patients with NTM-LD and may identify patients at risk of disease progression. A larger longitudinal study is needed to further evaluate this novel immunoprofiling approach.


Mycobacterium Infections, Nontuberculous , Pneumonia , Humans , Pilot Projects , Prospective Studies , Leukocytes, Mononuclear , Mycobacterium Infections, Nontuberculous/microbiology , Nontuberculous Mycobacteria
12.
BMC Infect Dis ; 24(1): 437, 2024 Apr 24.
Article En | MEDLINE | ID: mdl-38658840

BACKGROUND: Immunodeficient patients, particularly HIV patients, are at risk of opportunistic infections. Nontuberculous mycobacteria can cause severe complications in immunodeficient patients. CASE PRESENTATION: We describe a 57-year-old HIV patient, primarily presented with coughs and constitutional symptoms, with a unique Mycobacterium genavense abdominal, pulmonary, and central nervous system infection, accompanied by intracranial masses. CONCLUSION: The diagnosis of NTM, including M. genavense, must always be considered by clinicians in immunodeficient patients, especially those with HIV, who have a compromised immune system.


HIV Infections , Mycobacterium Infections, Nontuberculous , Humans , Middle Aged , HIV Infections/complications , Mycobacterium Infections, Nontuberculous/microbiology , Mycobacterium Infections, Nontuberculous/drug therapy , Male , Nontuberculous Mycobacteria/isolation & purification , Mycobacterium/isolation & purification , AIDS-Related Opportunistic Infections/microbiology , AIDS-Related Opportunistic Infections/drug therapy , AIDS-Related Opportunistic Infections/diagnosis
13.
Rev Esp Quimioter ; 37(3): 266-269, 2024 Jun.
Article En | MEDLINE | ID: mdl-38602224

OBJECTIVE: Mycobacterium avium complex (MAC) and Mycobacterium abscessus are a group of nontuberculous mycobacteria (NTM) that have been described as human pathogens. Their ability to develop biofilms in tissues and medical devices is one of the most important pathogenicity factors, with important implications in diagnosis and treatment. Macrolides are usually considered one of the bases of this treatment. METHODS: Here we have studied the biofilm prevention concentration (BPC) of 16 strains (n=16) with clarithromycin to avoid the biofilm development by these NTM. RESULTS: In this study, all M. abscessus strains have similar BPC, while MAC strains showed different values. For MAC the concentrations ranged between 1-16 mg/L, while for M. abscessus the concentration was 32 mg/L for all strains except one that was 64 mg/L. CONCLUSIONS: These results open the possibility of using macrolides for the prevention of biofilm development in patients with a risk of developing NTM disease.


Anti-Bacterial Agents , Biofilms , Clarithromycin , Microbial Sensitivity Tests , Nontuberculous Mycobacteria , Clarithromycin/pharmacology , Biofilms/drug effects , Anti-Bacterial Agents/pharmacology , Nontuberculous Mycobacteria/drug effects , Humans , Mycobacterium Infections, Nontuberculous/microbiology , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium Infections, Nontuberculous/prevention & control , Mycobacterium avium Complex/drug effects , Mycobacterium abscessus/drug effects
14.
J Infect ; 88(5): 106149, 2024 May.
Article En | MEDLINE | ID: mdl-38574774

BACKGROUND: Distinguishing between nontuberculous mycobacterial (NTM) lung infections and pulmonary tuberculosis becomes challenging due to their similar clinical manifestations and radiological images. Consequently, instances of delayed diagnosis or misdiagnosis are highly frequent. A feasible and reliable indicator of the existence of NTM in the early stages of the disease would help to solve this dilemma. METHODS: In this study, we evaluated the potential of smear-positive and Xpert assay (Cepheid, USA) negative outcomes as an early indicator of possible NTM infection in a high TB-burden setting retrospectively and prospectively. RESULTS: During the study period, 12·77% (138/1081) of the smear-positive cases yielded negative outcomes with the simultaneous Xpert assay. From the 110 patients who yielded smear-positive/Xpert-negative outcomes and cultivated strain as well, 105 (95·45%) were proved to have NTM isolated. By incorporating an additional criterion of a negative result from the Interferon-gamma release assay, the accuracy of the screening method reached 100%. Regarding the NTM presence prediction value, smear-positive/Xpert-negative has a sensitivity of 24·86% (45/181) in all NTM isolated cases but 93·75-96·55% accuracy in retrospective study or 93·75% accuracy in prospective study in smear-positive NTM isolated cases. In addition, the specificity was ∼99·47% (943/948) in smear-positive tuberculosis cases. CONCLUSION: The clue of the presence of NTM could be obtained on the first day of the hospital visit due to the point of care (POC) feature of smear testing and Xpert assay. About one-fourth of the NTM-isolated patients would benefit from this rapid, convenient, and reliable screening strategy in the given circumstance. Smear-positive/Xpert-negative outcome is an early, trustable indicator that is indicative of NTM isolation.


Mycobacterium Infections, Nontuberculous , Nontuberculous Mycobacteria , Sensitivity and Specificity , Humans , Mycobacterium Infections, Nontuberculous/diagnosis , Mycobacterium Infections, Nontuberculous/microbiology , Male , Female , Retrospective Studies , Nontuberculous Mycobacteria/isolation & purification , Nontuberculous Mycobacteria/genetics , Middle Aged , Prospective Studies , Aged , Adult , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/microbiology , Sputum/microbiology , Interferon-gamma Release Tests/methods , Diagnosis, Differential , Aged, 80 and over
15.
Antimicrob Agents Chemother ; 68(5): e0017424, 2024 May 02.
Article En | MEDLINE | ID: mdl-38557171

Mycobacterium abscessus (MAB) infections pose a growing public health threat. Here, we assessed the in vitro activity of the boronic acid-based ß-lactamase inhibitor, vaborbactam, with different ß-lactams against 100 clinical MAB isolates. Enhanced activity was observed with meropenem and ceftaroline with vaborbactam (1- and >4-fold MIC50/90 reduction). CRISPRi-mediated blaMAB gene knockdown showed a fourfold MIC reduction to ceftaroline but not the other ß-lactams. Our findings demonstrate vaborbactam's potential in combination therapy against MAB infections.


Anti-Bacterial Agents , Boronic Acids , Cefoxitin , Ceftaroline , Cephalosporins , Imipenem , Meropenem , Microbial Sensitivity Tests , Mycobacterium abscessus , Mycobacterium abscessus/drug effects , Meropenem/pharmacology , Boronic Acids/pharmacology , Anti-Bacterial Agents/pharmacology , Cephalosporins/pharmacology , Imipenem/pharmacology , Cefoxitin/pharmacology , Humans , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium Infections, Nontuberculous/microbiology , beta-Lactamase Inhibitors/pharmacology
16.
Microbiol Spectr ; 12(5): e0000624, 2024 May 02.
Article En | MEDLINE | ID: mdl-38619253

Mycobacterium abscessus is increasingly recognized as an emerging opportunistic pathogen causing severe lung diseases and cutaneous infections. However, treatment of M. abscessus infections remains particularly challenging, largely due to intrinsic resistance to a wide panel of antimicrobial agents. New therapeutic alternatives are urgently needed. Herein, we show that, upon limited irradiation with a blue-light source, newly developed porphyrin-peptide cage-type photosensitizers exert a strong bactericidal activity against smooth and rough variants of M. abscessus in planktonic cultures and in biofilms, at low concentrations. Atomic force microscopy unraveled important morphological alterations that include a wrinkled and irregular bacterial surface. The potential of these compounds for a photo-therapeutic use to treat M. abscessus skin infections requires further evaluations.IMPORTANCEMycobacterium abscessus causes persistent infections and is extremely difficult to eradicate. Despite intensive chemotherapy, treatment success rates remain very low. Thus, given the unsatisfactory performances of the current regimens, more effective therapeutic alternatives are needed. In this study, we evaluated the activity of newly described porphyrin-peptide cage-type conjugates in the context of photodynamic therapy. We show that upon light irradiation, these compounds were highly bactericidal against M. abscessus in vitro, thus qualifying these compounds for future studies dedicated to photo-therapeutic applications against M. abscessus skin infections.


Anti-Bacterial Agents , Biofilms , Microbial Sensitivity Tests , Mycobacterium Infections, Nontuberculous , Mycobacterium abscessus , Photosensitizing Agents , Porphyrins , Mycobacterium abscessus/drug effects , Porphyrins/pharmacology , Porphyrins/chemistry , Photosensitizing Agents/pharmacology , Photosensitizing Agents/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Biofilms/drug effects , Humans , Mycobacterium Infections, Nontuberculous/microbiology , Mycobacterium Infections, Nontuberculous/drug therapy , Peptides/pharmacology , Peptides/chemistry , Photochemotherapy/methods , Light
17.
Microbiol Spectr ; 12(5): e0383723, 2024 May 02.
Article En | MEDLINE | ID: mdl-38619262

Mycobacterium abscessus, an emerging pathogen responsible for severe pulmonary infections in cystic fibrosis patients, displays either a smooth (S) or a rough (R) morphotype. Infections with M. abscessus R are associated with increased pathogenicity in animal models and humans. While the S-to-R transition correlating with reduced glycopeptidolipid (GPL) production is well-documented, the recent screening of a transposon library revealed additional gene candidates located outside of the GPL locus involved in this transition. These genes include MAB_1470c, encoding the putative lipoprotein peptidase LpqM. However, experimental confirmation of the implication of this gene in the morphotype switch is lacking. Herein, we re-examined the role of MAB_1470c, and its homolog MAB_1466c, in colonial morphotype changes by generating unmarked deletion mutants in M. abscessus S. Our results indicate that the morphotype of these mutants stayed smooth in different media. Unexpectedly, the intracellular growth of ΔMAB_1470c and ΔMAB_1466c in THP-1 macrophages was significantly reduced as compared to the parental S strain, and these defects were rescued upon complementation with their corresponding genes. Strikingly, the intracellular survival defect was further exacerbated in a mutant lacking both MAB_1470c and MAB_1466c genes. This implies that, despite their primary sequence relatedness, the two proteins are not functionally redundant. Collectively, this suggests that these two LpqM-related lipoproteins are unlikely to be involved in the S-to-R transition but are key players for intramacrophage survival of M. abscessus. IMPORTANCE: Mycobacterium abscessus causes persistent infections in patients with underlying pulmonary diseases, resulting in progressive lung function deterioration. The rough (R) morphotype is well-established as associated with chronic and more aggressive infections in patients. In this study, we individually and simultaneously deleted the MAB_1470c and MAB_1466c genes in M. abscessus S, without observing changes in colony morphotypes. However, these mutants exhibited a severe impairment in their ability to survive within human macrophages, highlighting the critical role of these two lipoproteins in M. abscessus virulence.


Bacterial Proteins , Macrophages , Mycobacterium Infections, Nontuberculous , Mycobacterium abscessus , Mycobacterium abscessus/genetics , Mycobacterium abscessus/metabolism , Mycobacterium abscessus/growth & development , Humans , Macrophages/microbiology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Mycobacterium Infections, Nontuberculous/microbiology , THP-1 Cells , Virulence/genetics
18.
Clin Microbiol Infect ; 30(6): 738-742, 2024 Jun.
Article En | MEDLINE | ID: mdl-38527611

BACKGROUND: Antibiotic treatment of Mycobacterium abscessus disease is toxic and poorly effective and lacks a firm evidence base. Dual ß-lactam and ß-lactam/ß-lactamase inhibitor combinations may be interesting leads to improve treatment outcomes. OBJECTIVES: To summarize the current preclinical studies on dual ß-lactam and ß-lactam/ß-lactamase inhibitor combinations against M. abscessus. SOURCES: We performed a literature search using the National Center for Biotechnology Information's PubMed interface with additional snowball sampling. CONTENT: Select combinations of ß-lactam antibiotics, as well as ß-lactam/ß-lactamase inhibitor combinations show promising in vitro activity and synergy against M. abscessus. ß-Lactam antibiotics differ in their ability to reach and interfere with their targets and their resistance to the M. abscessus ß-lactamase. The synergy is typically observed for combinations of ß-lactam antibiotics or a ß-lactam antibiotic with a ß-lactamase inhibitor. No additional killing capacity was demonstrated in three-drug combinations of synergistic ß-lactam antibiotics and a ß-lactamase inhibitor. The efficacy of select dual ß-lactam antibiotics and ß-lactam/ß-lactamase inhibitor combinations is retained in intracellular infection assays and mouse models, but no combination has a complete preclinical portfolio. IMPLICATIONS: Future clinical strategies should entail either dual ß-lactam or ß-lactam/ß-lactamase inhibitor combinations. Imipenem-ceftaroline and an all-oral tebipenem-avibactam combination are promising leads but still require a complete preclinical portfolio, target product profiles as well as clinical trial confirmation.


Anti-Bacterial Agents , Drug Synergism , Drug Therapy, Combination , Mycobacterium Infections, Nontuberculous , Mycobacterium abscessus , beta-Lactamase Inhibitors , beta-Lactams , Mycobacterium abscessus/drug effects , beta-Lactams/therapeutic use , beta-Lactams/pharmacology , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium Infections, Nontuberculous/microbiology , beta-Lactamase Inhibitors/therapeutic use , beta-Lactamase Inhibitors/pharmacology , Humans , Treatment Outcome , Animals , Microbial Sensitivity Tests
19.
BMC Microbiol ; 24(1): 100, 2024 Mar 26.
Article En | MEDLINE | ID: mdl-38532357

BACKGROUND: Cameroon is a tuberculosis (TB) burden country with a 12% positivity among TB presumptive cases. Of the presumptive cases with a negative TB test, some are infected with Non-tuberculous Mycobacteria (NTM). However, the diagnosis of NTM infections remains difficult due to the lack of tools in many laboratories, particularly in resource limited laboratories and remote setting. The present study was undertaken to determine NTM profile and associated comorbidities among TB presumptive people. METHODS: A retrospective study was conducted from December 2018 to December 2019 in the Tuberculosis-National Reference Laboratory (TB-NRL) for Bacteriological analysis of samples and Jamot Hospital of Yaounde (JHY) for clinical evaluation of confirmed NTM patients. We included in this study data of 5267 TB presumptive people previously diagnosed using three consecutive samples and having culture and SD Bioline results with or without Microscopy and reverse hybridization-based Line Probe Assay(LPA) results. The data on co-morbidities or history of people infected with NTM were then collected from the three participants with available clinical data. RESULTS: We collected data of 5267 presumptive TB people. Among them, 3436 (65.24%), have a positive culture with 3200 (60.75%) isolates belong to Mycobacterium tuberculosis Complex (MBTC) and 236 (4.48%) to NTM. Our results showed that, 123 (52.11%) NTM were isolated from people with negative microscopy and 113 (47.88%) from people with positive microscopy. Among the 236 NTM, 108 (45.8%) isolates were identified using LPA. M. fortuitum was the most represented species (32.41%) followed by M. intracellulare (19.44%). Sputum had the highest proportion of NTM (56%), followed by bronchial aspirations (31%). The extra-pulmonary samples presented lower proportions of isolates compared to pulmonary samples. Some patients affected with NTM presented comorbidities as HIV infection, Pulmonary tuberculosis, Type 2 diabetes, Chronic bronchitis and Alveolar pneumonia. CONCLUSIONS: Our study showed the presence of NTM strains among presumptive TB people with a predominance of M. fortuitum and M. intracellulare. It is important to implement a surveillance system of NTM in TB burden country and also to develop a point-of-care test for NTM identification in limited-resource settings.


Diabetes Mellitus, Type 2 , HIV Infections , Mycobacterium Infections, Nontuberculous , Tuberculosis , Humans , Nontuberculous Mycobacteria , HIV Infections/microbiology , Mycobacterium Infections, Nontuberculous/microbiology , Retrospective Studies , Cameroon , Tuberculosis/microbiology
20.
Article Zh | MEDLINE | ID: mdl-38538244

Non-tuberculosis mycobacterium (NTM) refers to a general term for a large group of mycobacteria, excluding the mycobacterium tuberculosis and mycobacterium leprae, which is an opportunistic pathogen. NTM pulmonary disease and pulmonary tuberculosis have very similar clinical and imaging manifestations. Ordinary sputum tests can not distinguish between mycobacterium tuberculosis and NTM accurately, and it needs to be differentiated through detection methods such as mycobacterium culture medium, high-performance liquid chromatography, and molecular biology. During the diagnosis of occupational pneumoconiosis, a sandblasting and polishing worker's lung CT showed dynamic changes in infiltrating shadows and cavities in the right lung. A sputum drug sensitivity test showed NTM infection, but the patient refused treatment. After 20 months, the CT examination of the lung showed further enlargement of infiltrating shadows and cavities, and NTM bacterial identification showed intracellular mycobacterial infection. Amikacin, moxifloxacin, azithromycin, and ethambutol combined antibacterial treatment were given. Currently, the patient is still under treatment.


Mycobacterium Infections, Nontuberculous , Mycobacterium tuberculosis , Silicosis , Tuberculosis, Pulmonary , Humans , Mycobacterium Infections, Nontuberculous/complications , Mycobacterium Infections, Nontuberculous/diagnosis , Mycobacterium Infections, Nontuberculous/microbiology , Tuberculosis, Pulmonary/complications , Nontuberculous Mycobacteria , Silicosis/complications
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