Eight weeks of high-intensity interval training alters the tongue microbiome and impacts nitrate and nitrite levels in previously sedentary men

Nitric oxide (∗NO) is a key signalling molecule, produced enzymatically via ∗NO synthases (NOS) or following the stepwise reduction of nitrate to nitrite via oral bacteria. Exercise training upregulates NOS expression and improves systemic health, but its effect on oral health, and more particularly the oral microbiome, has not been investigated. We used an exercise training study design to investigate changes in the tongue dorsum microbiome, and in nitrate and nitrite levels in the saliva, plasma and muscle, before, during and after an exercise training period. Eleven untrained males (age 25 ± 5 years, mass 64.0 ± 11.2 kg, stature 171 ± 6 cm, V ˙ O 2peak 2.25 ± 0.42 l min −1 ) underwent 8-weeks of high-intensity interval training (HIIT), followed by 12-weeks of detraining. The tongue dorsum microbiome was examined using Pac-Bio long-read 16S rRNA sequencing. Nitrate and nitrite levels were quantified with high-performance liquid chromatography. Grouped nitrite-producing species did not change between any timepoints. However, HIIT led to changes in the microbiome composition, increasing the relative abundance of some, but not all, nitrite-producing species. These changes included a decrease in the relative abundance of nitrite-producing Rothia and a decrease in Neisseria , alongside changes in 6 other bacteria at the genus level (all p ≤ 0.05). At the species level, the abundance of 9 bacteria increased post-training (all p ≤ 0.05), 5 of which have nitrite-producing capacity, including Rothia mucilaginosa and Streptococcus salivarius . Post-detraining, 6 nitrite-producing species remained elevated relative to baseline. Nitrate increased in plasma ( p = 0.03) following training. Nitrite increased in the saliva after training ( p = 0.02) but decreased in plasma ( p = 0.03) and muscle ( p = 0.002). High-intensity exercise training increased the abundance of several nitrite-producing bacteria and altered nitrate and nitrite levels in saliva, plasma, and muscle. Post-detraining, several nitrite-producing bacteria remained elevated relative to baseline, but no significant differences were detected in nitrate or nitrite levels. Switching from a sedentary to an active lifestyle alters both the microbiome of the tongue and the bioavailability of nitrate and nitrite, with potential implications for oral and systemic health. • Eight weeks of exercise training increases the concentration of salivary nitrite. • Plasma nitrate also increased, while nitrite levels in the plasma and muscle decreased. • None of these changes in nitrate and nitrite persisted following 12 weeks of detraining. • The relative abundance of 9 bacterial species increased following training. • Five, including Rothia mucilaginosa and Streptococcus salivarius have nitrite-producing capacity.