Purpose: To evaluate the knowledge and practice of the American Heart Association (AHA) 2007 guidelines among dentists in Hyderabad city, India, for the prevention of infective endocarditis (IE). Materials and Methods: A survey questionnaire was mailed to the dentists to assess their knowledge with regard to the antibiotic prophylaxis needed for specific cardiac conditions and the prophylaxis reasonable before various dental procedures in high-risk patients and the first-line and second-line antibiotic regimen prescribed. Results: Of the 190 registered dentists, 169 (88.94%) completed the questionnaire. The mean age of the population was 39.17 ± 8.23 years. The majority of the respondents correctly identified that having prosthetic cardiac valves, a previous history of IE illness and cardiac transplantation with subsequent cardiac valvulopathy are cardiac conditions requiring antibiotic prophylaxis. Prescription of first-line antibiotic drugs and second-line drug regimens corresponding to the current guidelines was correctly answered by only 56.21% dentists and 60.95% dentists, respectively. Conclusion: A relatively low level of knowledge of the new guidelines was reported among dentists, reflecting the need for more continuing dental education programmes. Keywords: antibiotic prophylaxis, dental procedures, dentists, infective endocarditis