Tuberculosis: New Aspects of an Old Disease

Tuberculosis is an ancient infectious disease that remains a threat for public health around the world. Although the etiological agent as well as tuberculosis pathogenesis is well known, the molecular mechanisms underlying the host defense to the bacilli remain elusive. In this paper we focus on the innate immunity of this disease reviewing well-established and consensual mechanisms like Mycobacterium tuberculosis interference with phagosome maturation, less consensual mechanism like nitric oxide production, and new mechanisms, such as mycobacteria translocation to the cytosol, autophagy, and apoptosis/necrosis proposed mainly during the last decade

Biofilms on health and disease

Summary: Biofilms; Microorganism identification; In vitro biofilm assembly; Biofilm carachterization; Biofilm phenotype / bacteria genome.N/

Brief introduction to microscopy techniques and to the course

Compare fundamental and operating principles of transmission electron microscopy (TEM), scanning electron microscopy (SEM) and light microscopy for different applications. The main topics discussed during the course will be introduced.N/

Exploring the advantages of using CryoEM in health-related projects

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Bacterial biofilms, antibiotic resistance and healthcare-associated infections: a dangerous connection

In 2012, were estimated 6.7 million cases of healthcare-associated infections (HAI) either in long-term care facilities or acute-care hospitals from which result 37,000 deaths configuring a serious public health problem. The etiological agents are diverse and often resistant to antimicrobial drugs. One of the mechanisms responsible for the emergence of drug resistance is biofilm assembly. Biofilms are defined as thin layers of microorganisms adhering to the surface of a structure, which may be organic or inorganic, together with the polymers that they secrete. They are dynamic structures which experience different stages of organization with the ageing and are linked to an increase in bacterial resistance to host defense mechanisms, antibiotics, sterilization procedures other than autoclaving, persistence in water distribution systems and other surfaces. The understanding of bacteria organization within the biofilm and the identification of differences between planktonic and sessile forms of bacteria will be a step forward to fight HAIs

Comparative Analysis of Two Candida parapsilosis Isolates Originating from the Same Patient Harbouring the Y132F and R398I Mutations in the ERG11 Gene

This article belongs to the Special Issue Cellular and Molecular Mechanisms of Multiple Drug Resistance (MDR)This work presents a comparative analysis of two clinical isolates of C. parapsilosis, isolated from haemoculture (HC) and central venous catheter (CVC). Both strains harboured Y132F and R398I mutations in the gene ERG11 associated with resistance to fluconazole (FLC). Differences between the HC and CVC isolates were addressed in terms of virulence, resistance to FLC, and lipid distribution. Expression of the ERG6 and ERG9 genes, lipid analysis, fatty acid composition, and lipase activity were assessed via qPCR, thin-layer chromatography/high-performance liquid chromatography, gas chromatography, and spectrophotometry, respectively. Regulation of the ERG6 and ERG9 genes did not prove any impact on FLC resistance. Analysis of lipid metabolism showed a higher accumulation of lanosterol in both the isolates regardless of FLC presence. Additionally, a decreased level of triacylglycerols (TAG) with an impact on the composition of total fatty acids (FA) was observed for both isolates. The direct impact of the ERG11 mutations on lipid/FA analysis has not been confirmed. The higher lipase activity observed for C. parapsilosis HC isolate could be correlated with the significantly decreased level of TAG. The very close relatedness between both the isolates suggests that one isolate was derived from another after the initial infection of the host.This research was funded by the Slovak Research and Development Agency under contracts of SK-PT-18-0006 as part of the Bilateral Cooperation Program (2019–2022), APVV-21-0302 and grant VEGA 2/0036/22 from the Ministry of Education, Science, Research, and the Sport of the Slovak Republic.info:eu-repo/semantics/publishedVersio

Environmental biofilms, consequences for health and disease

Ornamental waters of easy access and populated with animals are quite attractive and can hide threats to human health. Here we evaluated, during one year, the microbiota of ornamental waters in a Lisbon park in order to assess the risk for human health. A total of 26 different bacterial species were identified during the study. Among these are potential human pathogens such as Aeromonas, E. coli, K. pneumoniae, Pseudomonas and Raoutella. Aeromonas sobria is a resident bacterium whereas other species such as A. hydrophila and A. veronii were sporadically identified. A possible explanation for this result is the presence of a fish population. The presence of E. coli indicates water contamination with fecal materials. We first isolate this bacterium in June when the water temperature increased and then after it was a resident despite the natural temperature drop in the autumn. This might be explained by the replace of the fish population by ducks in October which lead to an increase in water tubirdity and total carbon content. Pseudomonas fluorescens, P. putida and P. oryzihabitans have been considered low-virulence bacteria but recently have been identified as ethiological agents of healthcare assotiated infections (HAI) mostly in immunosupressed individuals. Another recognized HAI agent present is K. pneumoniae. The same bacteria species were isolated from biofilm and water suggesting the existence of an equilibrium between planktonic and biofilm organized bacteria. The role played by biofilm assembly on the emergence of antibiotic resistance is still ongoing for the isolated bacteria, as well as the possible interaction between water chemical composition and bacteria isolates. Potential pathogens (biofilm or planktonic) represent a significant cause of acute bacterial infections mainly in specific populations (paediatric and geriatric). These results support a periodic control of ornamental water microbiota as simple preventive measure to avoid potential health issues.info:eu-repo/semantics/publishedVersio

Effect of Quorum Sensing Molecule Farnesol on Mixed Biofilms of Candida albicans and Staphylococcus aureus

This article belongs to the Special Issue Microbial Biofilms, Antimicrobials, and Virulence Determinants.The natural bioactive molecule farnesol (FAR) is widely studied mainly for its antibiofilm and antimicrobial properties. In addition, it increases the effectiveness of some antimicrobial substances, which makes it interesting for the development of combined therapy. In the present work, the effect of FAR either alone or in combination with oxacillin (OXA) on mixed biofilms formed by clinically relevant pathogens, Candida albicans and Staphylococcus aureus, was studied. S. aureus isolates used for biofilm formation originated from blood cultures and central venous catheters (CVC) were characterized in terms of antimicrobial resistance. The minimal biofilm inhibitory concentration (MBIC50) for FAR of 48 h mixed biofilms formed by the C. albicans and methicillin-sensitive S. aureus (MSSA) was determined to be 125 M, and for the mixed biofilms with methicillin-resistant S. aureus (MRSA) was determined to be 250 M. Treatment of mixed biofilms with OXA (2 mg/mL) showed 4% inhibition; however, the combination of OXA (2 mg/mL) and FAR (300 M) resulted in 80% inhibition of biofilms. In addition, planktonic cells of S. aureus exhibited an increased susceptibility to OXA, cefoxitin and kanamycin in the presence of FAR (150 and 300 M). Scanning electron microscopy (SEM) micrographs confirmed patchy biofilm and lack of candidal hyphae in the samples treated with FAR and FAR/OXA in comparison to control and mixed biofilms treated only with OXA. Intriguingly, in a pilot experiment using fluorescence in situ hybridization (FISH), considerable differences in activity (as indicated by ribosome content) of staphylococcal cells were detected. While the activity rate of the staphylococci in mixed biofilms treated with FAR was high, no FISH-positive signal for staphylococcal cells was found in the biofilm treated with FAR/OXA.This research was funded by the Slovak Research and Development Agency under contracts of SK-PT-18-0006 as part of the Bilateral Cooperation Program (2019–2022), APVV-21-0302 and APVV-18-0075. This work was also supported by the EU Grant number 952398—CEMBO, Call: H2020-WIDESPREAD-05-2020—Twinning.info:eu-repo/semantics/publishedVersio

Adenoid bacterial colonization in a paediatric population

Adenoids play a key role in both respiratory and ear infection in children. It has also been shown that adenoidectomy improves these symptoms in this population. The main goal of the present study was to evaluate adenoid bacterial colonization and document a possible relation with infectious respiratory disease. A prospective observational study was designed to evaluate the proposed hypothesis in a paediatric population submitted to adenoidectomy by either infectious or non-infectious indications and compare these two cohorts. A total of 62 patients with ages ranging from 1 to 12 years old were enrolled in the study. Adenoid surface, adenoid core and middle meatus microbiota were compared. A close association between adenoid colonization and nasal infection was found, supporting that adenoids may function as bacterial reservoir for upper airway infection. The obtained results also contribute to explain the success of adenoidectomy in patients with infectious indications.Nacional de Saúde Dr. Ricardo Jorge (Grant 2015DDI1143)info:eu-repo/semantics/publishedVersio