Functional soft liners have come in to practice with the incorporation of antimicrobial agent to reduce the microbial load. The present study was aimed to analyze and compare three different anti microbial agents i.e Carvacrol Oil, Ozonated Oil and Povidone iodine against Staphylococcus Aureus and Candida Albicans at three different time intervals of 24 hrs, 14 days and 1 month. Material & methods: A total of two hundred and forty soft liner discs were prepared using 3D printed circular rings of 10mm x 2mm dimensions. Sixty discs were made for each group (n=60) which were further subdivided into the subgroup (a) and subgroup (b), (n=30) to be tested against Candida albicans and Staphylococcus aureus for the study Group C (Control Group) was devoid of any antimicrobial agent, Group 1(Carvacrol oil ) was incorporated with 20 micron litres of carvacrol oil using a micro pipette, Group 2 (Ozonated oil) was incorporated with 10 micron litres of 5 % ozonated oil using a micro pipette and Group 3 (Povidone Iodine) was incorporated with 20 micron litres of 5 % Povidone Iodine using a micro pipette. Results: All the three antimicrobial agents against Staphylococcus Aureus, Povidone Iodine after 24 hrs had the highest zone of inhibition i.e 3.805+/- 1.205 cm which declines rapidly during the follow up whereas Carvacrol Oil had moderate zone of inhibition formation after 24 hrs i.e 2.6200 +/- 1.2127 cm which relatively stays constant in the follow up period after 1 month i.e 2.3500 +/- 0.91196 cm. Ozonated oil has the poorest antimicrobial response in the all three time frames. Conclusion:. The results show that against both Staphylococcus Aureus and Candida Albicans, Carvacrol Oil has the most profound and lasting antimicrobial action followed by Povidone Iodine and Ozonated oil. However, due to limited literature further research is required to study the in vivo response of these antimicrobial agents.