Biology subjects

Trampuz, A.

Publications and source records attributed to Trampuz, A..

3 recordsLinked to original sources

bioRxiv2023

Towards faster identification of MRSA and MSSA: analysis of calorimetric curve parameters from large hospital bacterial collections

There is a need to develop faster yet precise diagnostic tools for the identification of infectious agents and their levels of antimicrobial susceptibility. One such tool is calorimetry, as previous studies show that bacteria display unique signatures of calorimetric/metabolic activity that such signatures can be used for pathogen identification under controlled laboratory conditions. However, the level of variation of these unique signatures remains largely unexplored. Here, we use real-time isothermal microcalorimetry to disentangle the inter- and intra-species metabolic differences between S. aureus and S. epidermidis. We worked with a large collection of bacterial isolates obtained from patients with prosthetic joint infections as well as reference laboratory strains. We analyzed their calorimetric profiles and decomposed the curve parameters derived from them. As a result, we identified six calorimetric parameters that are useful for identification at the intra-species level, with a particular focus on MRSA. Furthermore, we found important differences between the thermograms of ATCC laboratory strains when compared against model thermograms generated from all our clinical isolates. These results indicate that accounting for metabolic variability can impact diagnosis precision. Finally, we discuss our findings and suggest ways to optimize calorimetric diagnostics and treatment approaches.

microbiology↗

bioRxiv2019

Chemical biofilm dislodgement with chelating and reducing agents in comparison to sonication: implications for the diagnosis of implant associated infection

Sonication of removed devices improved the microbiological diagnosis of infection. Recently, chemical agents have been investigated for dislodgement of biofilms, including the chelating agent ethylenediaminetetraacetic acid (EDTA) and the reducing agent dithiothreitol (DTT). We compared the efficacy of chemical methods (EDTA and DTT) to sonication for biofilm dislodgement. Staphylococcus epidermidis (ATCC 35984) and Pseudomonas aeruginosa (ATCC 53278) biofilms were grown on porous glass beads for 3 days. After biofilm formation, beads were exposed to 0.9% saline, sonication and/or chemical agents. Quantitative and qualitative biofilm analyses were performed by colony counting (CFU/ml), isothermal microcalorimetry and scanning electron microscopy. The colony counts after treatment with EDTA and DTT were similar to those after exposure to 0.9% saline (6.3, 6.1 and 6.0 log CFU/ml, respectively) for S. epidermidis biofilms, and (5.1, 5.2 and 5.0 log CFU/ml, respectively) for P. aeruginosa biofilm. Sonication detected higher CFU counts (7.5 log CFU/ml) for S. epidermidis; (p<0.05) and 6.5 log for P. aeruginosa biofilm (p <0.05). Concordant results were detected with isothermal microcalorimetry and scanning electron microscopy. In conclusion, the CFU count after treatment of S. epidermidis or P. aeruginosa biofilms with EDTA and DTT was similar to those observed after 0.9% saline used as control. In contrast, sonication was superior to chemical methods for biofilm dislodgment and detection of microorganisms in sonication fluid. In conclusion, our study showed that sonication is superior to chemical method to dislodge bacterial biofilm from artificial surface and should be considered as standard diagnostic method for biofilm detection in implant-associated infections.

microbiology↗

bioRxiv2018

Evaluation of Sb-1 bacteriophage activity in enhancing antibiotic efficacy against biofilm, degrading the exopolysaccharide matrix and targeting persister cells of Staphylococcus aureus

Most research on phage therapy focused on planktonic bacteria, whereas bacteriophage activity against biofilms is limited. We evaluated the capability of Staphylococcus aureus-specific bacteriophage Sb-1 to eradicate biofilm alone and in combination with different classes of antibiotics, to degrade the extracellular matrix and target persister cells. Biofilm of methicillin-resistant S. aureus (MRSA) ATCC 43300 was treated with Sb-1 alone or in (simultaneous or staggered) combination with either fosfomycin, rifampin, vancomycin, daptomycin or ciprofloxacin. The matrix was visualized by confocal fluorescent microscopy. Persister cells were treated with 104 and 107 PFU/mL Sb-1 for 3 hours in PBS, followed by CFU counting. Alternatively, bacteria were washed and incubated in fresh BHI medium and the bacterial growth assessed after further 24-hours. Pre-treatment with Sb-1 followed by the administration of sub-inhibitory concentrations of antibiotic exerted a considerable synergistic effect in eradicating MRSA biofilm. Sb-1 determined a dose-dependent reduction of matrix exopolysaccharide. 107 PFU/mL Sb-1 showed direct killing activity on persisters. However, even a lower titer had lytic activity when phage-treated persister cells were inoculated in fresh medium, reverting to a normal-growing phenotype. This study provides valuable data regarding the capability of Sb-1 to enhance antibiotic efficacy, exhibiting specific antibiofilm features. Its ability to degrade the MRSA polysaccharide matrix and target persister cells makes Sb-1 suitable for the therapy of biofilm-associated infections.

microbiology↗