Search

Invasive fungal diseases are common complications in critically ill patients and in those with significant underlying imbalanced immune systems. Fungal co-, and/or super-infections are emerging and have become a rising concern within the last few years. In Europe, cases of candidiasis and aspergillosis dominate, followed by mucormycosis in India. Epidemiological studies show an increasing trend in the incidence of all three entities. Parallel to this, a shift within the underlying fungal pathogens is observed. More non-albicans Candida infections and aspergillosis with cryptic species are on the rise; cryptic species may cover intrinsic resistance to azoles and other antifungal drugs. The recent COVID-19 pandemic led to a significantly increasing incidence of invasive fungal diseases among hospitalized patients., Competing Interests: Declaration of competing interest The authors declare no conflict of interest with this manuscript., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The rare, but emerging mold Aspergillus terreus is an important pathogen in some geographical areas, like Tyrol (Austria) and Houston (Texas). The reason for this high prevalence is unknown. The present serosurveillance study aimed to evaluate the trends in levels of A. terreus -specific IgG antibodies in various regions of Tyrol and to compare the results to the environmental spread of A. terreus in Tyrol. Therefore, 1058 serum samples from healthy blood donors were evaluated. Data revealed a significant difference between the Tyrolean Upland and Lowland. Moreover, female participants had higher A. terreus IgG antibody levels than male participants. The differences found in our study are consistent with the distributional differences in environmental and clinical samples described in previous studies, supporting that A. terreus IgG antibody levels reflect the environmental epidemiology of A. terreus in Tyrol.

Background: Invasive fungal infections are an important cause of morbidity and mortality in a broad range of patients. Adequate and early diagnosis is a challenge and of importance for improved survival. New molecular-based diagnostic methods are trendsetting, yet with the drawback that conventional tests receive less attention, in the laboratory as well as in the clinical setting., Objectives: We aimed to provide a useful recommendation for direct microscopy for effectively managing numerous specimens related to fungal infections, mainly covering opportunistic pathogens., Sources: A PubMed literature search covering direct fungal microscopy was performed with no restrictions on publication dates., Content: Best practise recommendations targeting the role of direct microscopy in diagnosing fungal infections are given. This review highlights when to perform direct microscopy, displays the main fungal morphologies, discusses the pitfalls related to microscopy, and recommends how to best report the results to clinicians., Implication: In many samples, the performance of direct microscopy provides an important diagnostic benefit that is greater than culture alone. Fluorescent dyes improve sensitivity and allow a fast and rapid read. Reporting includes the presence or absence of yeast forms, septate or non-septate hyphae, pigmentation, cellular location, or any other specific structures being present. The visualization of fungal elements from a sterile body site is proof of an infection, independent of other test reports., (Copyright © 2023 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Objective: In Friedreich's ataxia (FRDA), the most affected tissues are not accessible to sampling and available transcriptomic findings originate from blood-derived cells and animal models. Herein, we aimed at dissecting for the first time the pathophysiology of FRDA by means of RNA-sequencing in an affected tissue sampled in vivo., Methods: Skeletal muscle biopsies were collected from seven FRDA patients before and after treatment with recombinant human Erythropoietin (rhuEPO) within a clinical trial. Total RNA extraction, 3'-mRNA library preparation and sequencing were performed according to standard procedures. We tested for differential gene expression with DESeq2 and performed gene set enrichment analysis with respect to control subjects., Results: FRDA transcriptomes showed 1873 genes differentially expressed from controls. Two main signatures emerged: (1) a global downregulation of the mitochondrial transcriptome as well as of ribosome/translational machinery and (2) an upregulation of genes related to transcription and chromatin regulation, especially of repressor terms. Downregulation of the mitochondrial transcriptome was more profound than previously shown in other cellular systems. Furthermore, we observed in FRDA patients a marked upregulation of leptin, the master regulator of energy homeostasis. RhuEPO treatment further enhanced leptin expression., Interpretation: Our findings reflect a double hit in the pathophysiology of FRDA: a transcriptional/translational issue and a profound mitochondrial failure downstream. Leptin upregulation in the skeletal muscle in FRDA may represent a compensatory mechanism of mitochondrial dysfunction, which is amenable to pharmacological boosting. Skeletal muscle transcriptomics is a valuable biomarker to monitor therapeutic interventions in FRDA., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The use of bacteriophages for the treatment of bacterial infections has been extensively studied. Nonetheless, the stress response regarding bacteriophage infection and the expression of virulence factors of Pseudomonas aeruginosa after phage infection is poorly discussed. In this study, we evaluated biofilm formation capacity and expression of virulence factors of P. aeruginosa after bacteriophage infection. Biofilm growth rates, biofilm morphology, pyocyanin production and elastase activity were evaluated after 2, 8, 24 and 48 h of co-cultivation with bacteriophages that was recently characterized and showed to be infective towards clinical isolates. In parallel, quantitative real-time polymerase chain reactions were carried out to verify the expression of virulence-related genes. Bacteriophages promoted substantial changes in P. aeruginosa biofilm growth at early co-culture time. In addition, at 8 h, we observed that some cultures developed filaments. Although bacteriophages did not alter both pyocyanin and protease activity, changes on the expression level of genes related to virulence factors were detected. Usually, lasI, pslA, lasB and phzH genes were upregulated after 2 and 48 h of co-culture. These results highlight the need for extensive investigation of pathways and molecules involved in phage infection, since the transcriptional changes would suggest a response activation by P. aeruginosa., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Implantable medical devices, such as prosthetics, catheters, and several other devices, have revolutionized medicine, but they increase the infection risk. In previous decades, commercially available antibiotics lost their activity against coagulase-negative Staphylococci (CoNS) and several other microorganisms. Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) are the two major omega-3 polyunsaturated fatty acids (ω-3 PUFAs) with antimicrobial properties. Materials and Methods: In this study, we tested the EPA and the DHA for its antibacterial and anti-biofilm activity in vitro against Staphylococcus epidermidis , Staphylococcus aureus , and different CoNS as reference strains and isolated from patients undergoing orthopedic treatment for implant infections. The tests were carried out with the strains in planktonic and biofilm form. Cytotoxicity assay was carried out with EPA and DHA using human gingival fibroblasts HGF-1. Results: The highest concentration of EPA and DHA promoted the complete killing of S. epidermidis 1457 and S. aureus ATCC 25923 in planktonic form. The fatty acids showed low activity against P. aeruginosa . EPA and DHA completely killed or significantly reduced the count of planktonic bacteria of the patient isolated strains. When incubated with media enriched with EPA and DHA, the biofilm formation was significantly reduced on S. epidermidis 1457 and not present on S. aureus ATCC 25923. The reduction or complete killing were also observed with the clinical isolates. The pre-formed biofilms showed reduction of the cell counting after treatment with EPA and DHA. Conclusion: In this study, the ω-3 PUFAs EPA and DHA showed antimicrobial and anti-biofilm activity in vitro against S. aureus , S. epidermidis , and P. aeruginosa , as well as against multi-drug resistant S. aureus and CoNS strains isolated from patients undergoing periprosthetic joint infections (PJI) treatment. Higher concentrations of the fatty acids showed killing activity on planktonic cells and inhibitory activity of biofilm formation. Although both substances showed antimicrobial activity, EPA showed better results in comparison with DHA. In addition, when applied on human gingival fibroblasts in vitro, EPA and DHA showed a possible protective effect on cells cultured in medium enriched with ethanol. Further studies are required to confirm the antimicrobial activity of EPA and DHA against multi-drug resistant strains and pan-drug resistant strains.

Objectives: For a better understanding of the mechanisms involved in biofilm formation, we performed a broad identification and characterization of the strains affecting implants by evaluating the morphology of biofilms formed in vitro in correlation with tests of the strains' antibiotic susceptibility in planktonic form. The ability of the strains to form biofilms in vitro was evaluated by means of colony forming units counting, metabolic activity tests of biofilm cells, and scanning electron microscopy. Methods: A total of 140 strains were isolated from patients with orthopedic implant-related infections during the period of 2015 to 2018. The identification of the isolates was carried out through microbiological cultures and confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Antibiotic susceptibility rates of the isolates were accessed according to EUCAST (European Committee on Antimicrobial Susceptibility Testing). The ability of all isolates to form biofilms in vitro was evaluated by counting the colony forming units, by measuring the metabolic activity of biofilm cells, and by analyzing the morphology of the formed biofilms using scanning electron microscopy. Results : From all the isolates, 41.84% (62 strains) were Staphylococcus epidermidis and 15.60% (22 strains) were Staphylococcus aureus. A significant difference in the capacity of biofilm formation was observed among the isolates. When correlating the biofilm forming capacity of the isolates to their antibiotic susceptibility rates, we observed that not all strains that were classified as resistant were biofilm producers in vitro. In other words, bacteria that are not good biofilm formers can show increased tolerance to multiple antibiotic substances. Conclusion : From 2015 until 2018, Staphylococcus epidermidis was the strain that caused most of the orthopedic implant-related infections in our hospital . Not all strains causing infection in orthopedic implants are able to form biofilms under in vitro conditions. Differences were observed in the number of cells and morphology of the biofilms. In addition, antibiotic resistance is not directly related to the capacity of the strains to form biofilms in vitro. Further studies should consider the use of in vitro culture conditions that better reproduce the joint environment and the growth of biofilms in humans.

Objectives: The ability of certain bacteria to form biofilms underlies their capacity to cause medical device-associated infections. Most bacteria need the metal iron for their proliferation but also to form biofilms. The aim of this in vitro study was to investigate whether iron restriction upon application of the iron chelator deferiprone (DFP) impacts on bacterial biofilm formation and whether such an intervention can exert synergistic effects towards the antibacterial activity of three antibiotic compounds against coagulase-negative staphylococci (CNS) residing on titanium plates., Methods: Bacteria were seeded on titanium discs and cultured to obtain biofilms. Biofilms were then exposed to DFP and/or antibiotic treatment with clindamycin, gentamycin or vancomycin. Fluorescence microscopy and scanning electron microscopy (SEM) were used for morphological analysis of the biofilms before and after treatment., Results: Whereas DFP alone had only a moderate inhibitory effect on biofilm growth, the combination of DFP with the respective antibiotics resulted in a significant decline of bacterial numbers by two to three logs as compared to the effect of antibiotics alone. Fluorescence staining and SEM demonstrated severe damage to even complete destruction of biofilms after combined treatment with DFP and antibiotics that was not the case upon sole treatment with antibiotics., Conclusion: Iron chelation is able to potentiate the antibacterial activity of conventional antibiotics by destroying bacterial biofilms that recommends this combination as a promising strategy for the treatment of chronic device infections with biofilm producing CNS.