Genome-Scale Characterization of Mycobacterium abscessus Complex Isolates from Portugal.

The Mycobacterium abscessus complex (MABC) is an emerging, difficult to treat, multidrug-resistant nontuberculous mycobacteria responsible for a wide spectrum of infections and associated with an increasing number of cases worldwide. Dominant circulating clones (DCCs) of MABC have been genetically identified as groups of strains associated with higher prevalence, higher levels of antimicrobial resistance, and worse clinical outcomes. To date, little is known about the genomic characteristics of MABC species circulating in Portugal. Here, we examined the genetic diversity and antimicrobial resistance profiles of 30 MABC strains isolated between 2014 and 2022 in Portugal. The genetic diversity of circulating MABC strains was assessed through a gene-by-gene approach (wgMLST), allowing their subspecies differentiation and the classification of isolates into DCCs. Antimicrobial resistance profiles were defined using phenotypic, molecular, and genomic approaches. The majority of isolates were resistant to at least two antimicrobials, although a poor correlation between phenotype and genotype data was observed. Portuguese genomes were highly diverse, and data suggest the existence of MABC lineages with potential international circulation or cross-border transmission. This study highlights the genetic diversity and antimicrobial resistance profile of circulating MABC isolates in Portugal while representing the first step towards the implementation of a genomic-based surveillance system for MABC at the Portuguese NIH.

Microevolution of a Mycobacteroides abscessus subsp. bolletii strain in a clinical persistent infection.

Mycobacteroides abscessus complex (MAB), a fast-growing nontuberculous mycobacterium, is emerging as a significant infectious disease threat, due to both intrinsic and acquired resistance mechanisms to antibiotics and disinfectants and the need for extensive and multidrug regimens for treatment. Despite the prolonged regimens, outcomes are poor and persistence cases have been reported. Here, we describe clinical, microbiologic and genomic features of a M. abscessus subsp. bolletii (M. bolletii) strain consecutively isolated from a patient within an eight-year infection period. From April 2014 to September 2021, the National Reference Laboratory for Mycobacteria received eight strains isolated from a male patient. Species identification, molecular resistance profile and phenotypic drug susceptibility were determined. Five of these isolates were recovered for further in-depth genomic analysis. Genomic analysis confirmed the multidrug resistant pattern of the strain and also other genetic changes associated with adaptation to environment and defence mechanisms. We highlight the identification of new mutations in locus MAB_1881c and in locus MAB_4099c (mps1 gene), already described as associated with macrolides resistance and morphotype switching, respectively. Additionally, we also observed the emergence and fixation of a mutation in locus MAB_0364c that appeared at a frequency of 36% for the 2014 isolate, 57% for the 2015 isolate and 100% for the 2017 and 2021 isolates, clearly illustrating a fixation process underlying a microevolution of the MAB strain within the patient. Altogether these results suggest that the observed genetic alterations are a reflection of the bacterial population's continuous adaptation and survival to the host environment during infection, contributing to persistence and treatment failure.

Molecular Capture of Mycobacterium tuberculosis Genomes Directly from Clinical Samples: A Potential Backup Approach for Epidemiological and Drug Susceptibility Inferences.

The application of whole genome sequencing of Mycobacterium tuberculosis directly on clinical samples has been investigated as a means to avoid the time-consuming need for culture isolation that can lead to a potential prolonged suboptimal antibiotic treatment. We aimed to provide a proof-of-concept regarding the application of the molecular capture of M. tuberculosis genomes directly from positive sputum samples as an approach for epidemiological and drug susceptibility predictions. Smear-positive sputum samples (n = 100) were subjected to the SureSelectXT HS Target Enrichment protocol (Agilent Technologies, Santa Clara, CA, USA) and whole-genome sequencing analysis. A higher number of reads on target were obtained for higher smear grades samples (i.e., 3+ followed by 2+). Moreover, 37 out of 100 samples showed ≥90% of the reference genome covered with at least 10-fold depth of coverage (27, 9, and 1 samples were 3+, 2+, and 1+, respectively). Regarding drug-resistance/susceptibility prediction, for 42 samples, ≥90% of the >9000 hits that are surveyed by TB-profiler were detected. Our results demonstrated that M. tuberculosis genome capture and sequencing directly from clinical samples constitute a potential valid backup approach for phylogenetic inferences and resistance prediction, essentially in settings when culture is not routinely performed or for samples that fail to grow.

Diagnostic Images of Pulmonary Alveolar Microlithiasis: A Rare, Autosomal Recessive Disorder.

Pulmonary alveolar microlithiasis (PAM) is a genetic lung disorder that is characterized by the accumulation of calcium phosphate deposits in the alveolar spaces of the lung. PAM is discovered incidentally on radiographs performed for other purposes, and the typical disease course is characterized by slowly progressive respiratory failure over decades. Treatment remains supportive. A 62-year-old woman presented in the emergency department with dyspnoea and fatigue. On physical examination she had crackles on pulmonary auscultation and digital clubbing. A CT scan of the chest showed multiple high-density areas throughout the lung parenchyma, suggesting the presence of alveolar microlithiasis. This CT finding is the typical radiological presentation of PAM, while the hallmark presentation is clinical-radiological dissociation. LEARNING POINTS: Pulmonary alveolar microlithiasis (PAM) is a rare genetic lung disorder resulting in accumulation of calcium phosphate deposits in the alveoli.The typical radiological presentation of PAM is the classic 'sandstorm' appearance in the lung.The key to diagnosis of this disease is clinical-radiological dissociation.

Mass Spectrometry-Based Proteomic and Metabolomic Profiling of Serum Samples for Discovery and Validation of Tuberculosis Diagnostic Biomarker Signature.

Tuberculosis (TB) is a transmissible disease listed as one of the 10 leading causes of death worldwide (10 million infected in 2019). A swift and precise diagnosis is essential to forestall its transmission, for which the discovery of effective diagnostic biomarkers is crucial. In this study, we aimed to discover molecular biomarkers for the early diagnosis of tuberculosis. Two independent cohorts comprising 29 and 34 subjects were assayed by proteomics, and 49 were included for metabolomic analysis. All subjects were arranged into three experimental groups­healthy controls (controls), latent TB infection (LTBI), and TB patients. LC-MS/MS blood serum protein and metabolite levels were submitted to univariate, multivariate, and ROC analysis. From the 149 proteins quantified in the discovery set, 25 were found to be differentially abundant between controls and TB patients. The AUC, specificity, and sensitivity, determined by ROC statistical analysis of the model composed of four of these proteins considering both proteomic sets, were 0.96, 93%, and 91%, respectively. The five metabolites (9-methyluric acid, indole-3-lactic acid, trans-3-indoleacrylic acid, hexanoylglycine, and N-acetyl-L-leucine) that better discriminate the control and TB patient groups (VIP > 1.75) from a total of 92 metabolites quantified in both ionization modes were submitted to ROC analysis. An AUC = 1 was determined, with all samples being correctly assigned to the respective experimental group. An integrated ROC analysis enrolling one protein and four metabolites was also performed for the common control and TB patients in the proteomic and metabolomic groups. This combined signature correctly assigned the 12 controls and 12 patients used only for prediction (AUC = 1, specificity = 100%, and sensitivity = 100%). This multiomics approach revealed a biomarker signature for tuberculosis diagnosis that could be potentially used for developing a point-of-care diagnosis clinical test.

Uncovering Beta-Lactam Susceptibility Patterns in Clinical Isolates of Mycobacterium tuberculosis through Whole-Genome Sequencing.

The increasing threat of drug resistance and a stagnated pipeline of novel therapeutics endanger the eradication of tuberculosis. Beta-lactams constitute promising additions to the current therapeutic arsenal and two carbapenems are included in group C of medicines recommended by the WHO for use in longer multidrug-resistant tuberculosis regimens. However, the determinants underlining diverse Mycobacterium tuberculosis phenotypes to beta-lactams remain largely undefined. To decipher these, we present a proof-of-concept study based on a large-scale beta-lactam susceptibility screening for 172 M. tuberculosis clinical isolates from Portugal, including 72 antimycobacterial drug-resistant strains. MICs were determined for multiple beta-lactams and strains were subjected to whole-genome sequencing to identify core-genome single-nucleotide variant-based profiles. Global and cell wall-targeted approaches were then followed to detect putative drivers of beta-lactam response. We found that drug-resistant strains were more susceptible to beta-lactams, but significant differences were not observed between distinct drug-resistance profiles. Sublineage 4.3.4.2 strains were significantly more susceptible to beta-lactams, while the contrary was observed for Beijing and 4.1.2.1 sublineages. While mutations in beta-lactamase or cell wall biosynthesis genes were uncommon, a rise in beta-lactam MICs was detected in parallel with the accumulation of mutations in peptidoglycan cross-linking or cell division genes. Finally, we exposed that putative beta-lactam resistance markers occurred in genes for which relevant roles in cell wall processes have been ascribed, such as rpfC or pknA. Genetic studies to validate the relevance of the identified mutations for beta-lactam susceptibility and further improvement of the phenotype-genotype associations are needed in the future. IMPORTANCE Associations between differential M. tuberculosis beta-lactam phenotypes and preexisting antimycobacterial drug resistance, strain sublineage, or specific mutational patterns were established. Importantly, we reveal that highly drug-resistant isolates of sublineage 4.3.4.2 have an increased susceptibility to beta-lactams compared with other strains. Thus, directing beta-lactams to treat infections by specific M. tuberculosis strains and refraining its use from others emerges as a potentially important strategy to avoid resistance development. Individual mutations in blaC or genes encoding canonical beta-lactam targets, such as peptidoglycan transpeptidases, are infrequent and do not greatly impact the MICs of potent carbapenem plus clavulanic acid combinations. An improved understanding of the global effect of cumulative mutations in relevant gene sets for peptidoglycan and cell division processes on beta-lactam susceptibility is also provided.

[The role of enjoyment and motivational determinants in persistence in the practice of physical exercise]. / O papel do divertimento e das determinantes motivacionais na persistência da prática de exercício físico.

The scope of this paper was to analyze the impact of motivational determinants in enjoyment and persistence in physical exercise among practitioners of physical exercise. In total, 967 gym and health club exercise practitioners aged between 18 and 65 (M=45.08; SD=13.76) were recruited for analysis. All participants had more than six months of regular exercise practice. Participants completed and validated scales in the exercise context duly translated into Portuguese, assessing interpersonal behaviors, basic psychological needs, and behavioral regulation. Persistence was measured using computerized records considering persistent exercisers as being those who were exercising at similar frequencies as those self-reported at the initial assessment. Results showed that the measurement and structural model fit the data. Several significant effects were found supporting previous literature. Indirect effects showed enjoyment to play a crucial role on exercise persistence, both by the significant effect via autonomous and controlled motivation. In essence, activities that give pleasure that gym and health club exercisers experience during the practice of physical exercise can be the key variable of long-term persistence.

Como objetivo deste trabalho, definiu-se analisar os efeitos indiretos de determinantes motivacionais no divertimento e na persistência em praticantes de exercício físico regular. Participaram ao todo 967 praticantes de exercício em ginásio e health clubs, com idades compreendidas entre 18 e 65 anos (M=45,08; DP=13,76). Todos os participantes tinham mais de 6 meses de experiência regular em exercício físico. Os participantes preencheram ao todo questionários traduzidos e validados para a língua portuguesa no contexto do exercício físico, que examinavam a perceção dos comportamentos interpessoais, as necessidades psicológicas básicas, a regulação da motivação e o divertimento. A persistência foi medida através de registos eletrónicos, considerando um praticante persistente aquele que tivesse uma frequência semanal similar aquela auto-reportada no momento inicial. A maioria das regressões são significativas, confirmado os pressupostos da literatura existente. Os efeitos indiretos mostram que o divertimento desempenha um papel crucial na persistência, tanto por via motivação autónoma, como por via motivação controlada. Em suma, atividades promotoras do prazer que os praticantes de ginásio e health club experienciam durante a prática de exercício físico poderão ser a variável chave na persistência a longo prazo.

O papel do divertimento e das determinantes motivacionais na persistência da prática de exercício físico / The role of enjoyment and motivational determinants in persistence in the practice of physical exercise

Resumo Como objetivo deste trabalho, definiu-se analisar os efeitos indiretos de determinantes motivacionais no divertimento e na persistência em praticantes de exercício físico regular. Participaram ao todo 967 praticantes de exercício em ginásio e health clubs, com idades compreendidas entre 18 e 65 anos (M=45,08; DP=13,76). Todos os participantes tinham mais de 6 meses de experiência regular em exercício físico. Os participantes preencheram ao todo questionários traduzidos e validados para a língua portuguesa no contexto do exercício físico, que examinavam a perceção dos comportamentos interpessoais, as necessidades psicológicas básicas, a regulação da motivação e o divertimento. A persistência foi medida através de registos eletrónicos, considerando um praticante persistente aquele que tivesse uma frequência semanal similar aquela auto-reportada no momento inicial. A maioria das regressões são significativas, confirmado os pressupostos da literatura existente. Os efeitos indiretos mostram que o divertimento desempenha um papel crucial na persistência, tanto por via motivação autónoma, como por via motivação controlada. Em suma, atividades promotoras do prazer que os praticantes de ginásio e health club experienciam durante a prática de exercício físico poderão ser a variável chave na persistência a longo prazo.

Abstract The scope of this paper was to analyze the impact of motivational determinants in enjoyment and persistence in physical exercise among practitioners of physical exercise. In total, 967 gym and health club exercise practitioners aged between 18 and 65 (M=45.08; SD=13.76) were recruited for analysis. All participants had more than six months of regular exercise practice. Participants completed and validated scales in the exercise context duly translated into Portuguese, assessing interpersonal behaviors, basic psychological needs, and behavioral regulation. Persistence was measured using computerized records considering persistent exercisers as being those who were exercising at similar frequencies as those self-reported at the initial assessment. Results showed that the measurement and structural model fit the data. Several significant effects were found supporting previous literature. Indirect effects showed enjoyment to play a crucial role on exercise persistence, both by the significant effect via autonomous and controlled motivation. In essence, activities that give pleasure that gym and health club exercisers experience during the practice of physical exercise can be the key variable of long-term persistence.

The Co-Occurrence of Satisfaction and Frustration of Basic Psychological Needs and Its Relationship with Exercisers' Motivation.

Although the relationship between both need frustration and, particularly, need satisfaction and different motivational regulations for exercise has been widely examined in the literature, little is known about the co-occurrence of both need satisfaction and need frustration in the exercise context. Grounded in self-determination theory, the present study aimed to examine the effects of both need satisfaction and frustration on motivational regulations for exercise, by applying a response surface analysis approach. In total, 477 regular exercisers aged 18-54 years participated in this study. The interaction between needs (high on both need satisfaction and frustration) displayed a positive and significant association with amotivation, integrated regulation, and intrinsic motivation. Considering the direction of the discrepancy (high vs. low levels of need satisfaction and frustration) related to the behavioral regulations, results showed that higher need satisfaction relative to need frustration was associated with more self-determined regulations of motivation. Contrarily, higher need satisfaction relative to need frustration was associated with lower scores on amotivation, external, introjected, and identified regulation. Overall, these findings extend previous literature, suggesting that need satisfaction and frustration are distinct factors that can be experienced simultaneously in individuals during exercise and that different degrees of both needs have different associations with behavioral regulations.

Exercise Promotion: Reviewing the Importance of Health Professionals' Interpersonal Behaviors on Exercisers' Basic Psychological Needs.

Previous studies have investigated the impact of exercisers' perceptions of health professionals' interpersonal behaviors on exercisers' exercise adherence. From these studies, there is increased interest in developing and evaluating programs to improve health professionals' communication skills and interpersonal behavior. In this narrative review, we provide examples of self-determination theory and newer modifications to it, discuss the empirical conditions that foster optimal exerciser motivation, consider the antecedent factors influencing health professionals' behaviors, and offer practical suggestions to health professionals seeking to promote regular exercise practice. Since exercisers perceive and differentiate health professionals' need-supportive, need-thwarting, and need-passive behaviors, health professionals who can critically and consciously distinguish these different types of behavior are more likely to foster supportive climates and suppress the use of need-thwarting and need-indifferent behaviors. The interpersonal interaction between health professionals and exercisers strongly influences how exercisers will regulate their behavior toward persistent exercise.

Insights on the Mycobacterium tuberculosis population structure associated with migrants from Portuguese-speaking countries over a three-year period in Greater Lisbon, Portugal: Implications at the public health level.

Tuberculosis among foreign-born patients is a key indicator of country-level epidemiological profiles and, of an increasing concern in Europe given the more intensified migratory waves of refugees. Since Portugal presents a lower immigrant-associated TB incidence rate when compared to other European countries, we sought to characterize the epidemiology and transmission dynamics among the foreign-born population coming from Portuguese-speaking countries that are associated with higher TB incidences. In the present study we analyzed 133 Mycobacterium tuberculosis isolates obtained from foreign-born individuals over a three-year period in Lisbon, Portugal, using molecular epidemiological methods such as spoligotyping and 24-loci MIRU-VNTR. Moreover, all strains were subjected to drug susceptibility testing. The genetic profiles obtained suggest that strain importation from Portuguese speaking countries plays a less important role in TB epidemiology but instead argue in favor of a high degree of penetrance of Portuguese endemic strains to the migrant population, including multidrug resistant strains, which is particularly relevant to active screening programs.

Evaluation of a gene-by-gene approach for prospective whole-genome sequencing-based surveillance of multidrug resistant Mycobacterium tuberculosis.

Whole-genome sequencing (WGS) offers unprecedented resolution for tracking Mycobacterium tuberculosis transmission and antibiotic-resistance spread. Still, the establishment of standardized WGS-based pipelines and the definition of epidemiological clusters based on genetic relatedness are under discussion. We aimed to implement a dynamic gene-by-gene approach, fully relying on freely available software, for prospective WGS-based tuberculosis surveillance, demonstrating its application for detecting transmission chains by retrospectively analysing all M/XDR strains isolated in 2013-2017 in Portugal. We observed a good correlation between genetic relatedness and epidemiological links, with strongly epilinked clusters displaying mean pairwise allele differences (AD) always below 0.3% (ratio of mean AD over the total number of shared loci between same-cluster strains). This data parallels the genetic distances acquired by the core-SNV analysis, while providing higher resolution and epidemiological concordance than MIRU-VNTR genotyping. The dynamic analysis of strain sub-sets (i.e., increasing the number of shared loci within each sub-set) also strengthens the confidence in detecting epilinked clusters. This gene-by-gene strategy also offers several practical benefits (e.g., reliance on freely-available software, scalability and low computational requirements) that further consolidated its suitability for a timely and robust prospective WGS-based laboratory surveillance of M/XDR-TB cases.

Trends of multidrug-resistant tuberculosis clustering in Portugal.

Analysis of MDR-TB rates showed consistent decreases in cases and clustering rates but did not establish valid relationships with the epidemiological information collected by the public health authorities http://ow.ly/3BnN30nxqW3.

Go with the flow: advances and trends in magnetic flow cytometry.

The growing need for biological information at the single cell level has driven the development of improved cytometry technologies. Flow cytometry is a particularly powerful method that has evolved over the past few decades. Flow cytometers have become essential instruments in biomedical research and routine clinical tests for disease diagnosis, prognosis, and treatment monitoring. However, the increasing number of cellular parameters unveiled by genomic, proteomic, and metabolomic data platforms demands an augmented multiplexability. Also, the need for identification and quantification of relevant biomarkers at low levels requires outstanding analytical sensitivity and reliability. In addition, growing awareness of the advantages associated with miniaturization of analytical devices is pushing forward the progress in integrated and compact, microfluidic-based devices at the point-of-care. In this context, novel types of flow cytometers are emerging during the search to tackle these challenges. Notwithstanding the relevance of other promising alternatives to standard optical flow cytometry (e.g., mass cytometry, various optical and electrical microcytometers), this report focuses on a recent microcytometric technology based on magnetic sensors and magnetic particles integrated into microfluidic structures for dynamic bioanalysis of fluid samples-magnetic flow cytometry. Its concept, main developments, targeted applications, as well as the challenges and trends behind this technology are presented and discussed. Graphical abstract ᅟ "Kindly advise whether there is online abstract figure for this paper. If so, kindly resupply.The graphical abstract is correctly supplied.

Clonal expansion across the seas as seen through CPLP-TB database: A joint effort in cataloguing Mycobacterium tuberculosis genetic diversity in Portuguese-speaking countries.

Tuberculosis (TB) remains a major health problem within the Community of Portuguese Language Speaking Countries (CPLP). Despite the marked variation in TB incidence across its member-states and continued human migratory flux between countries, a considerable gap in the knowledge on the Mycobacterium tuberculosis population structure and strain circulation between the countries still exists. To address this, we have assembled and analysed the largest CPLP M. tuberculosis molecular and drug susceptibility dataset, comprised by a total of 1447 clinical isolates, including 423 multidrug-resistant isolates, from five CPLP countries. The data herein presented reinforces Latin American and Mediterranean (LAM) strains as the hallmark of M. tuberculosis populational structure in the CPLP coupled with country-specific differential prevalence of minor clades. Moreover, using high-resolution typing by 24-loci MIRU-VNTR, six cross-border genetic clusters were detected, thus supporting recent clonal expansion across the Lusophone space. To make this data available to the scientific community and public health authorities we developed CPLP-TB (available at http://cplp-tb.ff.ulisboa.pt), an online database coupled with web-based tools for exploratory data analysis. As a public health tool, it is expected to contribute to improved knowledge on the M. tuberculosis population structure and strain circulation within the CPLP, thus supporting the risk assessment of strain-specific trends.

Redox status on different regions of the central nervous system of obese and lean rats treated with green tea extract.

OBJECTIVES: The purpose of this study was to evaluate some indicators of redox status, and inflammation on different regions of the central nervous system (CNS) of obese rats treated with green tea (GT). We hypothesized that obesity could affect the redox balance in different brain regions due to the diverse nature of the cells as well as the selective neuronal vulnerability to oxidative stress, and GT could triggers benefits effects restoring the redox status. METHODS: Male Wistar rats were treated with GT by gavage (12 weeks/5 days/week; 500 mg/kg of body weight) and obesity was induced by cafeteria diet (8 weeks). After this period, the animals were killed and brain tissue (cerebral cortex, cerebellum, and brainstem) was removed to evaluate oxidative stress and inflammation (cytokine release). RESULTS: We showed that the cafeteria diet had little effect on redox balance in the cerebral cortex and cerebellum; however, the brainstem was the region of the CNS most sensitive to cafeteria diet-induced redox unbalance. GFAP expression was increased in the cerebral cortex of obese rats and reduced by GT. It was also evident that GT treatment had numerous beneficial effects against oxidative damage to biomolecules in all brain regions analyzed. DISCUSSION: Our study established that different CNS regions show selective neuronal vulnerability when exposed to a diet enriched with fats and sugars, and the beneficial effect of GT was similar among these regions. We conclude that GT could be a good strategy for improving and maintaining brain function under healthy and pathological conditions.

Portuguese in vitro antibiotic susceptibilities favor current nontuberculous mycobacteria treatment guidelines.

SETTING: Nontuberculous mycobacteria (NTM) are increasingly recognized as causative agents of opportunistic infections in humans for which effective treatment is challenging. There is, however, very little information on the prevalence of NTM drug resistance in Portugal. OBJECTIVE AND DESIGN: Our aim was to analyze the drug susceptibility testing (DST) performed in NTM at the Portuguese National Health Institute Dr. Ricardo Jorge from February 2003 to February 2016. A total of 262 DST were included in the analysis. RESULTS: Most (94%) M. avium intracellulare complex isolates showed in vitro susceptibility to clarithromycin. All M. kansasii isolates were susceptible to rifampicin and ethambutol and 97.1% were susceptible to isoniazid. The majority of rapidly-growing mycobacteria (RGM) demonstrated in vitro susceptibility to amikacin, clarithromycin and cefoxitin. However, in RGM there was a marked increase on the relative risk of having sulfamethoxazole resistance in isolates resistant to ciprofloxacin compared to susceptible isolates. CONCLUSION: Tested NTM in Portugal revealed in vitro susceptibility to most of the antimicrobials currently recommended for treatment. However, our results also suggest that sulfamethoxazole should be avoided in treatment of RGM resistant to ciprofloxacin (or vice versa). Further trials that correlate the in vitro DST results with the clinical outcome are needed in order to reach conclusions on efficient antimicrobial therapy.

Dissecting whole-genome sequencing-based online tools for predicting resistance in Mycobacterium tuberculosis: can we use them for clinical decision guidance?

Whole-genome sequencing (WGS)-based bioinformatics platforms for the rapid prediction of resistance will soon be implemented in the Tuberculosis (TB) laboratory, but their accuracy assessment still needs to be strengthened. Here, we fully-sequenced a total of 54 multidrug-resistant (MDR) and five susceptible TB strains and performed, for the first time, a simultaneous evaluation of the major four free online platforms (TB Profiler, PhyResSE, Mykrobe Predictor and TGS-TB). Overall, the sensitivity of resistance prediction ranged from 84.3% using Mykrobe predictor to 95.2% using TB profiler, while specificity was higher and homogeneous among platforms. TB profiler revealed the best performance robustness (sensitivity, specificity, PPV and NPV above 95%), followed by TGS-TB (all parameters above 90%). We also observed a few discrepancies between phenotype and genotype, where, in some cases, it was possible to pin-point some "candidate" mutations (e.g., in the rpsL promoter region) highlighting the need for their confirmation through mutagenesis assays and potential review of the anti-TB genetic databases. The rampant development of the bioinformatics algorithms and the tremendously reduced time-frame until the clinician may decide for a definitive and most effective treatment will certainly trigger the technological transition where WGS-based bioinformatics platforms could replace phenotypic drug susceptibility testing for TB.