An Anaerobic Environment Drives the Harboring of Helicobacter pylori within Candida Yeast Cells

SIMPLE SUMMARY: Helicobacter pylori is a pathogen that is associated with a number of gastric pathologies and has adapted to the gastric environment. Outside this organ, stress factors such as oxygen concentration affect the viability of this bacterium. This study aimed to determine if changes in oxygen concentration promoted the entry of H. pylori into the interior of yeast cells of the Candida genus. Co-cultures of H. pylori and Candida strains in Brucella broth plus 5% fetal bovine serum were incubated under microaerobic, anaerobic, or aerobic conditions. Bacteria-like bodies (BLBs) were detected within yeast cells (Y-BLBs) by optical microscopy, identified by molecular techniques, and their viability evaluated by SYTO-9 fluorescence. Co-cultures incubated under the three conditions showed the presence of Y-BLBs, but the highest Y-BLB percentage was present in H. pylori J99 and C. glabrata co-cultures incubated under anaerobiosis. Molecular techniques were used to identify BLBs as H. pylori and SYTO-9 fluorescence confirmed that this bacterium remained viable within yeast cells. In conclusion, although without apparent stress conditions H. pylori harbors within Candida yeast cells, its harboring increases significantly under anaerobic conditions. This endosymbiotic relationship also depends mostly on the H. pylori strain used in the co-culture. ABSTRACT: Helicobacter pylori protects itself from stressful environments by forming biofilms, changing its morphology, or invading eukaryotic cells, including yeast cells. There is little knowledge about the environmental factors that influence the endosymbiotic relationship between bacterium and yeasts. Here, we studied if oxygen availability stimulated the growth of H. pylori within Candida and if this was a bacterial- or yeast strain-dependent relationship. Four H. pylori strains and four Candida strains were co-cultured in Brucella broth plus 5% fetal bovine serum, and incubated under microaerobic, anaerobic, or aerobic conditions. Bacteria-like bodies (BLBs) within yeast cells (Y-BLBs) were detected by microscopy. H. pylori was identified by FISH and by PCR amplification of the 16S rRNA gene of H. pylori from total DNA extracted from Y-BLBs from H. pylori and Candida co-cultures. BLBs viability was confirmed by SYTO-9 fluorescence. Higher Y-BLB percentages were obtained under anaerobic conditions and using H. pylori J99 and C. glabrata combinations. Thus, the H. pylori–Candida endosymbiotic relationship is strain dependent. The FISH and PCR results identified BLBs as intracellular H. pylori. Conclusion: Stressful conditions such as an anaerobic environment significantly increased H. pylori growth within yeast cells, where it remained viable, and the bacterium–yeast endosymbiotic relationship was bacterial strain dependent with a preference for C. glabrata.