Contamination of cell cultures by mycoplasmas is a very common phenomenon. As they can substantially alter cell metabolism and potentially spread to all cell cultures in laboratory, their early detection is necessary. One of the fastest and cheapest methods of mycoplasma detection relies on the direct staining of mycoplasmas’ DNA by DAPI or Hoechst dyes. Although this method is easy and fast to perform, it suffers from the low signal provided by these dyes compared to the nuclear DNA. Therefore, the reporter cell lines are used for cultivation of mycoplasmas before DAPI or the Hoechst staining step. In the study presented, we have developed and tested a new immunofluorescence assay for the detection of mycoplasmas. The method is based on the enzymatic labeling using DNA polymerase I and modified nucleotides utilizing nicks in the mycoplasmas’ DNA. Modified nucleotides are incorporated into mycoplasmas’ DNA and subsequently visualized by immunofluorescence microscopy. The developed approach is independent of the mycoplasma strain, does not intensely stain nuclear DNA, does not stain other bacteria, and provides higher sensitivity than the approach based on the direct labeling using DAPI or Hoechst dyes.

A blend with pumpkin and sunflower seed flours was prepared and dried at 41.5 °C for 5 h to create a minimally heat-treated blend for a raw food diet. The blend was inoculated with Lactobacillus acidophilus and Fusarium langsethiae to assess the effect of L. acidophilus on Fusarium growth and mycotoxin production. Drying did not affect the content of naturally occurring microorganisms but significantly reduced water activity (p < 0.05) and increased total phenolic content in samples with external microorganisms. Lactobacilli content remained unchanged after drying (4.8 log CFU/g), while F. langsethiae increased by 1.5 log CFU/g. Principal component analysis showed PC1 explained 95.1% of total variance, driven by Fusarium mycotoxin production. A significant difference in total mycotoxin was found between samples with F. langsethiae alone and those with both F. langsethiae and L. acidophilus (p < 0.05). Lactic acid bacteria could reduce fusarium mycotoxin risk in raw food diet mixtures.

Hyaluronic acid (HA) is a natural biopolymer widely used in wound dressings for its supportive role in the healing process. In this study, we investigated the occurrence of microorganisms in chronic wounds and evaluated the antimicrobial activity of newly synthesized HA–Starch-based materials enriched with acidifying agents. Microbial isolates obtained from chronic wounds were tested for susceptibility using the agar diffusion method. The prepared materials exhibited significant antimicrobial activity against both reference strains and multidrug-resistant clinical isolates. Further characterization by scanning electron microscopy and elemental analysis confirmed uniform microfiber morphology and the expected elemental composition of the fibers. Cytotoxicity assessments performed using the xCELLigence system demonstrated the potential safety of developed materials. Overall, the results indicate that HA–Starch-based materials containing acidifying compounds exhibit strong in vitro antimicrobial activity against chronic-wound isolates, supporting their potential for further evaluation in wound care applications.