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To evaluate the ‘in vitro’ bactericidal effect of the Nd:YAG laser (Genius, MØlsgaard Dental, Copenhagen, Denmark) on six periodontal pathogens.Suspensions of six different periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum and Parvimonas micra) were prepared in small Eppendorff tubes, and exposed to a Nd:YAG laser for five different periods of time. Laser settings used: Power 6 Watt (on a scale of 1–12 W), Frequency 50 Hz, Pulse duration 250 μs. After exposure to the laser, aliquots of the suspensions were spread on blood agar plates for bacterial counting.After 5 s of laser exposure, there was a decrease in total colony forming units for all six selected microorganisms. After 15, 30 and 45 s, no viable bacterial cells could be retrieved.In this ‘in vitro’ model, 15 s of Nd:YAG laser use was found to be effective for total killing of the six tested periodontal pathogens.