Impact of Long-term Fasting on Breath Volatile Sulphur Compounds, Inflammatory Markers and Saliva Microbiota Composition

Background and Purpose: Despite substantial evidence supporting the role of resident bacterial communities in therapeutic fasting outcomes, research has primarily focused on gut microbiota, leaving changes in oral microbiota largely unexplored. The clinical significance of oral health changes during fasting is nonetheless underscored by the documented development of halitosis in fasting individuals. However, no scientific studies have comprehensively examined the interplay between salivary microbiota alterations, inflammatory changes in the gingival crevice, and the production of malodorous volatile compounds. We examined volatile sulphur compounds (VSC) in breath during fasting, cytokine levels in the gingival crevice, and oral microbiota composition of the saliva in a single-arm interventional study involving 36 subjects who fasted for 10 ± 3 days.
Materials and Methods: Participants fasted according to Buchinger fasting guidelines. VSC were evaluated every morning before any food or drink intake using the OralChroma gas chromatography device. Saliva and gingival crevicular fluid (GCF) samples were collected at the clinical site before fasting, at the end of fasting, and at the end of food reintroduction. Follow-up saliva samples were sent to the patients after 1 and 3 months. Saliva samples were processed and analysed by targeted sequencing of 16S rRNA gene amplicons, whereas the expression of 6 inflammatory markers in the GCF were analysed using a multiplex fluorescent bead-based immunoassay.
Results: The quantification of volatile compounds in the breath demonstrated a statistically significant increase in dimethylsulfide levels during fasting, which corroborates the occurrence of bad breath as a common side effect of fasting. Salivary microbiota profiling showed a shift in microbial composition, including reduction in the levels of Neisseria, Gemella and Porphyromonas spp., concomitant with an increase in the levels of Megasphaera, Dialister, Prevotella, Veillonella, Bifidobacteria, Leptotrichia, Selenomonas, Alloprevotella, and Atopobium. We further demonstrated a reduction in the levels of the pro-inflammatory cytokine interleukin-8 in the GCF.
Conclusion: Dimethylsulfide concentrations in the breath increased during fasting, and this was correlated to changes in the oral microbiota. Future studies are needed to illuminate the possible impact of these changes on oral and general health status.