Resuscitation of the Helicobacter pylori Coccoid Forms by Resuscitation Promoter Factor Obtained from Micrococcus
Didem Aktas1, Melahat Bagirova2, Adil M Allahverdiyev1
1Faculty of Chemical and Metallurgical Engineering Department of Bioengineering, Yildiz Technical University, Davutpasa Campus, Istanbul, Turkey.
Abstract:
Helicobacter pylori is a gram negative, spiral-shaped, and microaerophilic bacteria which can cause life-threatening diseases. It is known that more than 55% of the human population in the world is already infected by this bacterium. The traditional treatment of H. pylori infection consists of a combination of two or more antibiotics. However, H. pylori has evolved to turning its shape from spiral to coccoid form in the presence of antibiotics and this decreases the therapeutic efficacies of conventional antibiotic applications. Resuscitation promoter factor (RPF) is a protein secreted by Micrococcus luteus have significant resuscitation effects on some bacteria especially in the group of viable but non-culturable (VBNC) pathogens. However, there is no study in the literature investigating the resuscitation effects of RPF derived from M. luteus on H. pylori in order to change its form from coccoid to spiral. The purpose of this study is to investigate the resuscitation effect of RPF-containing metabolites isolated from M. luteus on the morphological transformation of H. pylori coccoid forms to spiral forms in order to increase their susceptibilities to antibiotic treatments. Rpf-containing metabolites were primarily obtained from M. luteus culture supernatants. H. pylori was exposed to five different conditions such as prolonged culture, incubation at + 4 °C, incubation at + 22 °C, cultivation in PBS and treatment with kanamycin in order to induce transformations of bacteria to coccoid forms. Induced H.pylori coccoids were characterized by inverted microscope, UV spectrophotometer, SEM imaging, and flow-cytometer. As a result, it was found that the most suitable condition for inducing coccoid forms was cultivation of bacteria with kanamycin. Followingly, different concentrations of RPF-containing metabolites were applied on H. pylori coccoids induced by kanamycin. For the first time in this study, it was determined that the Rpf-containing metabolites obtained from M. luteus demonstrated very high resuscitation effect on kanamycin-induced H. pylori coccoid forms. This new approach for resuscitation of H. pylori coccoids is thought to play an important role in increasing the treatment effectiveness of the conventional antibiotics against the infection.
Insights
Resuscitation promoter factor (RPF) from Micrococcus luteus can revert antibiotic-induced Helicobacter pylori coccoid forms back to spiral shapes. This finding may enhance antibiotic efficacy against H. pylori infections.
Area of Science:
- Microbiology
- Bacterial Pathogenesis
- Drug Discovery
Background:
- Helicobacter pylori infection affects over 55% of the global population.
- H. pylori adopts a coccoid form under antibiotic pressure, reducing treatment effectiveness.
- Micrococcus luteus secretes resuscitation promoter factor (RPF) with known effects on viable but non-culturable bacteria.
Purpose of the Study:
- To investigate the potential of RPF from M. luteus to induce morphological transformation of H. pylori coccoid forms back to spiral forms.
- To assess if this transformation can increase H. pylori susceptibility to conventional antibiotic treatments.
Main Methods:
- Induction of H. pylori coccoid forms using kanamycin, prolonged culture, and temperature variations.
- Isolation of RPF-containing metabolites from M. luteus culture supernatants.
- Characterization of induced coccoid forms using microscopy, spectrophotometry, SEM, and flow cytometry.
- Application of RPF-containing metabolites to H. pylori coccoids and assessment of morphological changes.
Main Results:
- Kanamycin treatment was identified as the most effective method for inducing H. pylori coccoid forms.
- RPF-containing metabolites from M. luteus demonstrated a significant resuscitation effect on kanamycin-induced H. pylori coccoids.
- Morphological transformation from coccoid to spiral forms was observed for the first time using this method.
Conclusions:
- RPF from M. luteus can effectively resuscitate H. pylori coccoid forms into their spiral state.
- This novel approach holds promise for improving the efficacy of current antibiotic therapies against H. pylori infections.
- Further research into RPF-mediated resuscitation could lead to new strategies for combating H. pylori.
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