To assess and compare the anti-microbial efficacy of 445nm and 970nm diode laser on mixed species biofilm of Aggregatibacter actinomycetemcomitans [A.a] and Porphyromonas gingivalis [P.g] cultured on machined pure titanium discs. A total of 65 machined pure titanium discs with no surface modifications with a 10-mm diameter and a 2-mm height were sterilized by autoclaving at 121°C for 15min and incubated with the commercially available bacterial strains ATCC(American Type Culture Collection- P.g 33277 and A.a 29522)mixture of Aggregatibacter actinomycetemcomitans(A.a) and Porphyromonas gingivalis(P.g).After a 2-week incubation period with the mixture of bacteria to develop a mixed species biofilm, the discs were divided into three groups: (1) no treatment (control), (2) 445nm laser (test), (3) 970nm laser (test). For each laser wavelength (445 and 970nm), the discs were exposed to 1.0W and 2.0W in continuous wave mode for the times points of 15, 30, and 60s. The antimicrobial efficacy was assessed by qPCR. A significant reduction in the levels of both species of bacteria was observed between control and the laser intervention groups. A higher efficacy for the 445nm diode laser against Porphyromonas gingivalis and a similar efficacy against Aggregatibacter actinomycetemcomitans was observed as compared to the 970nm group. 445nm wavelength represents a potential and effective laser wavelength which can be used for the management of peri-implant infection. The present study findings also need to be further validated through clinical interventional trials.