Serial collection method of dog saliva: Effects of different chemical stimulants on behaviour, volume and saliva composition

The objective of this study was to evaluate different chemical stimulants with different flavours such as acids (citric and acetic), sweet (sucrose) and salty (sodium chloride) applied to cotton rolls and compare their effects on the volume, pH and protein concentrations of the saliva collected and the behaviour of dogs during sampling management. As an additional objective, serum cortisol concentrations of saliva samples obtained with or without citric acid and with or without previous pH adjustment were compared. Five clinically healthy were randomly assigned to one of 5 treatments with cottons with different substances: 1) control, 2) citric acid, 3) acetic acid, 4) sodium chloride, 5) sucrose. Each dog received one treatment per day, and in 5 days, all dogs were tested with the five treatments. On each day, cottons were applied to dogs at times 0, 20, 40, 60 and 80 minutes. The cottons with citric acid generated more volume than the rest of the treatments (p<0.0001), and sodium chloride generated more volume than the control and acetic acid (p≤0.03). Cottons with citric acid generated lower pH of saliva than the rest of the treatments (p<0.0001). Cottons with acetic acid generated lower pH than control, sodium chloride and sucrose (p<0.0001). There were no differences in cortisol concentrations between the control samples and those obtained with citric acid, nor between these same samples with and without pH adjusted with buffer. The concentration of proteins in saliva and excitement degree did not change with treatment. Citric acid was more palatable than the rest of the treatments (p<0.0001). Sodium chloride and sucrose were more palatable than control (p<0.05). In conclusion, citric acid was the chemical stimulant that generated greater volume of saliva and greater palatability in dogs. Although the pH of the saliva obtained with citric acid was clearly acidic, its acidic pH did not affect the determination of cortisol by chemiluminescence or RIA. Sodium chloride and sucrose allowed to obtain high volumes of saliva and were more palatable than the control, which can be other interesting options to obtain saliva in case of not being able to use citric acid.