Introduction: The aim of the present study was to investigate the effect of mechanical irrigation in combination with mouthwash of antimicrobial agents on salivary bacterial counts. Materials and Methods: This study was performed with a randomized study employing a panel of 40 healthy volunteers (20 males and 20 females) between the age of 26 and 32 years. Volunteers were randomly put in one of four treatment groups. In the first group, 0.2 mL of non-stimulatory saliva was collected from every subjective person. Then, saliva was collected after rinsing with chlorhexidine (CHX) for 1 minute. In the second group, non-stimulatory saliva was collected, and then saliva was collected after rinsing with CHX and irrigation with saline. In the third and fourth groups, the same procedures as the first and second groups were performed with povidone iodine (PVI) instead of CHX. All of these samples were cultured for 48 hours aerobically. The reduction rates of colony-forming units (CFU) were calculated for each group. The reduction rate between each group was tested statistically using student t-test. Results: Using CHX in combination with saline irrigation showed a significant decrease of the salivary bacterial CFU when compared with only using CHX.(P<0.01) And using PVI with saline irrigation showed a little decrease of the CFU when compared with only using PVI, but there was no statistical significance.(P>0.01) Conclusion: It was concluded that the CHX or PVI used with saline irrigation made the salivary bacterial counts reduced more than when CHX or PVI was used alone as an oral antiseptic agent.