Periodontitis is the most frequent oral health problem in the world. The infection is primarily caused by anaerobic microorganisms. Metronidazole is the most commonly used drug to treat the infection but recently the anaerobes have shown the resistance to this drug. Therefore, the present study was undertaken to isolate and identify the anaerobes associated with periodontitis and study their susceptibility pattern to the Metronidazole. Total 90 samples were collected from chronic periodontitis cases. Anaerobes were isolated in 71% of periodontitis cases. Gram positive organisms were more predominantly isolated than Gram negative organisms. The Gram negative anaerobes were found to be 100% sensitive to Metronidazole while Gram positive anaerobes showed 8% resistance to Metronidazole.