The aim of this study was to evaluate the influence of unstimulated and stimulated saliva collection methods, as well as tooth brushing, on the secretion rate of salivary total protein, nitrite, total antioxidant capacity and alpha-amylase. Saliva of 14 healthy individuals were collected with stimulation using Salivette, Parafilm and chewing gum and without stimulation from spit with and without fluid accumulation, before and after oral hygiene. Total protein, nitrite, total antioxidant capacity and alpha-amylase concentration (sAA) were evaluated. The collection of saliva stimulated with Parafilm and chewing gum increased the salivary flow (1.5 ± 0.4 and 3.4 ± 0.7 mL/min, respectively) and the secretion rate of salivary total protein (1.0 ± 0.2 and 2.3 ± 0.5 mg/min, respectively). Also, chewing gum increases the salivary nitrite secretion (213 ± 58 nmol/min) and total antioxidant capacity (410 ± 47 nmol trolox eq/min). Interestingly, the unstimulated method without saliva accumulation prior to collection resulted in low sAA levels (23,531 ± 7979 pixel density). Furthermore, oral hygiene decreased salivary flow (1.3 ± 0.5 to 1.0 ± 0.4 mL/min), reduced the secretion rate of total protein (1.0 ± 0.5 to 0.6 ± 0.2 mg/min, p < .05) and increased sAA (13,159 ± 7114 to 20,075 ± 25,656 pixel density, p < .05). The type of stimulation can activate autonomous receptors responsible for the secretion and composition of saliva. Therefore, the evaluation of saliva collection methods and oral hygiene on salivary biomarkers is important for understanding and standardizing variations in salivary composition to strengthen the use of saliva as a diagnostic fluid.

The collection of samples of saliva is noninvasive and straightforward, which turns saliva into an ideal fluid for monitoring the adaptive response to training. Here, we investigated the response of the salivary proteins alpha-amylase (sAA), chromogranin A (sCgA), and the concentration of total protein (sTP) as well as salivary nitrite (sNO2) in relation to plasma catecholamines and plasma nitrite (pNO2), respectively. The variation in these markers was compared to the intensity and load of training during a 21-week training season in 12 elite swimmers. Overall, the salivary proteins tracked the concentration of plasma adrenaline and were inversely correlated with the training outcomes. No correlations were observed between sNO2 and pNO2. However, sNO2 correlated positively with the intensity and load of training. We argue that the decrease in sympathetic activity is responsible for the decrease in the concentration of proteins throughout the training season. Furthermore, the increase in nitrite is likely to reflect changes in hemodynamics and regulation of vascular tone. The association of the salivary markers with the training outcomes underlines their potential as noninvasive markers of training status in professional athletes.

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