Objective: This study was conducted to evaluate the seroprevalence of HIV, HBV, HCV, and Syphilis and Malaria among blood donors. The data generated will help the clinicians for judicious use of blood as well as awareness regarding the Transfusion transmitted infections. Research Design Methods: A total of 46,224 blood donors were screened during a period from April 2008 to October 2012, at blood bank, S.R.G. Hospital and Medical College Jhalawar - District, Rajasthan State. Results: Among these 22905 (49.55%) were voluntary donors and 25219 (54.58%) were replacement donors .Seropositivity for Human Immunodeficiency Virus (HIV) was 0.034%, Hepatitis B Virus (HBV) was 1.57%, Hepatitis C Virus (HCV) was 0.04%, Rapid plasma Reagin method (RPR) for syphilis was 0.019% and Malaria was 0.017% respectively. Conclusions: Infections are slightly more common among replacement donors compared to voluntary donors. There was a gradual decrease of Transfusion Transmitted Infections (TTIs) in blood donors over the years by reason of following of stringent blood donor selection criteria.