Objective: To evaluate antibiotic prescribing practices of dentists working in a major teaching hospital in Sudan, and to assess the need for establishing education programmes to enhance dentists' awareness of antibiotic use. Material and Methods: A cross-sectional study was carried out by distributing an anonymous self-administered questionnaire to a convenience sample of dental practitioners at major teaching hospital in Khartoum/Sudan. The questionnaire investigated knowledge and attitudes of participants in relation to antibiotic prescribing in dental practice. Results: One hundred out of 135 dental practitioners working at the hospital participated in the survey with 48 males and 52 females. Most of the participants (N=44) did not specify their preferable drug reference, while 32 participants specified British National Formulary, and the practice was significantly associated with a higher professional rank (p<0.001). Metronidazole was the most frequently cited antibiotic for dentofacial infections (N=73) and Acute Necrotizing Ulcerative Gingivitis (N=56), while amoxicillin was the most frequently cited antibiotic for dento-alveolar infections and infective endocarditis prophylaxis (N=88 and N=47, respectively). Conclusion: Undergraduate, as well as postgraduate courses, are required in Sudan to provide up-to-date orientation on antibiotic use, and to increase awareness of dentists about antibiotic prescribing.