Healthcare-associated infections (HAIs) are caused by nosocomial pathogens. HAIs have an immense impact not only on developing countries but also on highly developed parts of world. They are predominantly device-associated infections that are caused by the planktonic form of microorganisms as well as those organized in biofilms. This review elucidates the impact of HAIs, focusing on device-associated infections such as central line-associated bloodstream infection including catheter infection, catheter-associated urinary tract infection, ventilator-associated pneumonia, and surgical site infections. The most relevant microorganisms are mentioned in terms of their frequency of infection on medical devices. Standard care bundles, conventional therapy, and novel approaches against device-associated infections are briefly mentioned as well. This review concisely summarizes relevant and up-to-date information on HAIs and HAI-associated microorganisms and also provides a description of several useful approaches for tackling HAIs.

Determining the number of viable cells by calculating colony-forming units is time-consuming. The evaluation of mixed biofilms consisting of different species is particularly problematic. Therefore, the aim of this study was to optimize a molecular method—propidium monoazide quantitative polymerase chain reaction (PMA-qPCR) - for accurate and consistent differentiation between living and dead cells. In the practical experimental example, the number of genome copies representing living cells was determined in a mixed biofilm of Candida albicans-Staphylococcus aureus inhibited by photodynamic inactivation. Optimal conditions such as PMA concentration and the duration of light exposure, the optimization of DNA isolation from the mixed biofilm and standardization of PMA-qPCR parameters were tested prior to the main experiment. The genome copy number was calculated based on the known amount of gDNA in the qPCR reaction and the genome size of the respective microorganism. The results showed that PDI in the presence of 1 mM methylene blue decreased the total genome copy number from 1.65x10⁸ to 3.19x10⁷, and from 4.39x10⁷ to 1.91x10⁷ for S. aureus and C. albicans (p < 0.01), respectively. The main disadvantage is overestimation of the number of living cells represented by genome copy numbers. Such cells are unable to reproduce and grow (no vitality), and are continuously dying. On the other hand, PMA-qPCR determines the copy numbers of all microbial species, including a mix of eukaryotic yeasts and prokaryotic bacteria in a biofilm in one step, which is a great advantage.

Candida albicans and Staphylococcus aureus are opportunistic pathogens that cause life-threatening infections. This study focused on using photodynamic inactivation (PDI) to eliminate mixed biofilms of C. albicans–S. aureus formed on poly (urethane) (PU) discs functionalized with a nanocomposite layer containing phloxine B (PhB). Additionally, the effect of PDI on the ALS3 and HWP1 genes of C. albicans was examined in mixed biofilms. Spectral analysis showed a continuous release of PhB from the nanocomposite in Mueller–Hinton broth within 48 h, with a released amount of PhB < 5% of the total amount. The anti-biofilm effectiveness of the light-activated nanocomposite with PhB showed a reduction in the survival rate of biofilm cells to 0.35% and 31.79% for S. aureus and C. albicans, respectively, compared to the control biofilm on PU alone. Scanning electron microscopy images showed that the nanocomposite effectively reduced the colonization and growth of the mixed biofilm. While PDI reduced the regulation of the ALS3 gene, the HWP1 gene was upregulated. Nevertheless, the cell survival of the C. albicans–S. aureus biofilm was significantly reduced, showing great potential for the elimination of mixed biofilms.