Tuberculosis (TB) remains an impactful infectious disease, leading to millions of deaths every year. Mycobacterium tuberculosis causes the formation of granulomas, which will determine, through the host–pathogen relationship, if the infection will remain latent or evolve into active disease. Early TB diagnosis is life-saving, especially among immunocompromised individuals, and leads to proper treatment, preventing transmission. This review addresses different approaches to diagnosing TB, from traditional methods such as sputum smear microscopy to more advanced molecular techniques. Integrating these techniques, such as polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP), has significantly improved the sensitivity and specificity of M. tuberculosis identification. Additionally, exploring novel biomarkers and applying artificial intelligence in radiological imaging contribute to more accurate and rapid diagnosis. Furthermore, we discuss the challenges of existing diagnostic methods, including limitations in resource-limited settings and the emergence of drug-resistant strains. While the primary focus of this review is on TB diagnosis, we also briefly explore the challenges and strategies for diagnosing non-tuberculous mycobacteria (NTM). In conclusion, this review provides an overview of the current landscape of TB diagnostics, emphasizing the need for ongoing research and innovation. As the field evolves, it is crucial to ensure that these advancements are accessible and applicable in diverse healthcare settings to effectively combat tuberculosis worldwide.

Since ancient times, plants have been used in folk medicine to treat different diseases. Plants offer exceptional chemical diversity with a wide range of biological activities, and have therefore been the most promising sources for the discovery and development of drugs, including antimicrobial agents. This study reports the antibacterial effect of geraniol (GER), alone and in combination with biogenic silver nanoparticles (bioAgNPs), produced using the aqueous extract of Trichilia catigua bark, against planktonic and sessile cells of methicillin-resistant Staphylococcus aureus (MRSA), one of the main opportunistic and potentially fatal human pathogens. GER had a time-dependent bactericidal effect on planktonic cells, impairing the cell membrane integrity. In addition, GER inhibited the staphyloxanthin production, and molecular docking analyses supported the in silico affinity of GER to dehydrosqualene synthase (CrtM) and 4,4′-diaponeurosporen-aldehyde dehydrogenase (AldH), which are key enzymes within the pigment biosynthesis pathway in S. aureus. GER treatment increased the sensitivity of MRSA to hydrogen peroxide killing. GER displayed synergism with bioAgNPs against planktonic and sessile cells, inhibiting bacterial adhesion and the viability of biofilms formed on abiotic surfaces. MRSA planktonic and sessile cells treated with GER or GER/bioAgNPs displayed severe morphological and ultrastructural alterations. Notably, neither GER nor its combination caused in vitro and in vivo toxicity in mammalian cells and Galleria mellonella larvae, respectively. These findings suggest that the combination of GER/bioAgNPs may be a promising strategy to control MRSA infections.

Candida tropicalis is regarded as an opportunistic pathogen, causing diseases ranging from superficial infections to life-threatening disseminated infections. The ability of this yeast to form biofilms and develop resistance to antifungals represents a significant therapeutic challenge. Herein, the effect of geraniol (GER), alone and combined with fluconazole (FLZ), was evaluated in the planktonic and sessile cells of azole-resistant C. tropicalis. GER showed a time-dependent fungicidal effect on the planktonic cells, impairing the cell membrane integrity. Additionally, GER inhibited the rhodamine 6G efflux, and the molecular docking analyzes supported the binding affinity of GER to the C. tropicalis Cdr1 protein. GER exhibited a synergism with FLZ against the planktonic and sessile cells, inhibiting the adhesion of the yeast cells and the viability of the 48-h biofilms formed on abiotic surfaces. C. tropicalis biofilms treated with GER, alone or combined with FLZ, displayed morphological and ultrastructural alterations, including a decrease in the stacking layers and the presence of wilted cells. Moreover, neither GER alone nor combined with FLZ caused toxicity, and both treatments prolonged the survival of the Galleria mellonella larvae infected with azole-resistant C. tropicalis. These findings indicate that the combination of GER and FLZ may be a promising strategy to control azole-resistant C. tropicalis infections.

Candida species are the primary fungal pathogens of invasive infections associated with high morbidity and mortality. The identification of these microorganisms is critical for therapeutic management and control of hospital infection. Herein, assays targeting the Intergenic Spacer 2 (IGS2) and Internal Transcribed Spacer 1 (ITS1) from the rDNA locus were developed to differentiate Candida species. Based on consensus nucleotide sequences, specific primers and positive controls were designed, and standard PCR and real-time PCR (qPCR) assays were performed. All primers resulted in specific amplification of the molecular targets of each species with no amplifications of the negative template control. Furthermore, the primers were highly specific when tested with a range of fungal DNAs and no cross-reactivity was observed among Candida species. The assays presented a limit of detection (LoD) of 10 copies of positive control per reaction for all specific primers designed. Overall, our results showed that qPCR assays employing primers targeting the regions IGS2 and ITS1 completely differentiated between Candida albicans, Candida auris, Candida parapsilosis, Candida tropicalis, and Nakazeomyces glabratus, with great accuracy and no amplification of DNA from other fungal species.

Candida albicans remains the most common agent of candidiasis worldwide. This yeast is generally sensitive to most antifungals, however, the emergence of azole-resistant C. albicans has been reported. In addition, this microorganism can form biofilms on various surfaces, making it difficult to treat infections. In this study, the effect of secondary metabolites of Pseudomonas aeruginosa strain LV on planktonic and sessile cells of C. albicans, with different genotypes and susceptibility profile to fluconazole and voriconazole, was evaluated. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the semi-purified fraction F4a ranged from 1.56 to 6.25 μg/mL and 6.25 to 25 μg/mL, respectively. Fluopsin C appears to be the antifungal component of F4a. The semi-purified fraction and fluopsin C showed fungicidal activity, dose and time dependent. F4a caused severe damage to the morphology and ultrastructure of planktonic fungal cells, and significantly reduced the viability of 24-hour biofilms, with MIC for sessile cells from 12.5 to 25.0 μg/mL. However, cytotoxicity was detected in mammalian cells for F4a and fluopsin C at concentrations that showed antifungal activity. These results indicate that fluopsin C may be a prototype for the development of new antifungals for C. albicans.

Produtos naturais são substâncias bioativas obtidas de fontes naturais como plantas, microrganismos, algas e animais, que podem possuir diversas propriedades terapêuticas. No presente estudo é abordado os principais bioativos extraídos das fontes de produtos naturais, como os alcaloides, terpenos, terpenoides, polifenóis e fenilpropanoides; além dos seus métodos de extração, purificação, identificação e as análises químicas e biológicas. Essa abordagem permite entender e caracterizar os ativos para avaliar suas propriedades terapêuticas, e garantir sua aplicabilidade na medicina tradicional ou medicina moderna, como a fitoterapia e homeopatia. Para obtenção destes produtos, é importante destacar a necessidade de conservação da biodiversidade, propiciando a obtenção continua destes e de novos compostos bioativos. No entanto, a pesquisa enfrenta desafios, incluindo a complexidade da extração, a variabilidade dos compostos e a necessidade de validação clínica rigorosa. A interdisciplinaridade na pesquisa e a conservação ambiental são fundamentais para explorar os benefícios dos produtos naturais, combinando conhecimentos tradicionais e avanços científicos para inovar na saúde humana.