Objective Our study planned to investigate the current positivity rate and distribution of the serologic markers of TTIs among male blood donors of the White Nile state, Sudan. Results The overall reported seropositive cases of TTIs was 15.91%, and percentages of anti-Human immunodeficiency virus 1/2 (anti-HIV1/2), Hepatitis B virus surface antigen (HBVsAg), anti-Hepatitis C virus (anti-HCV), and anti-Treponema palladium (anti-T. palladium) were 2.61%, 5.57%, 1.40%, and 5.72%, respectively. Out of 10897 donors examined, 0.59% had a serological sign of multiple infections. Furthermore, the odds of testing positive for TTIs were higher in the 28–37 age group (OR: 2.620, 95% CI: 2.324–2.955) and lower in the 38–47 age group (OR: 0.671, 95% CI: 0.567–0.794) compared to individuals of 18–27 years old. Likewise, it is more in individuals of Kosti (OR: 1.122, 95% CI: 0.987–1.277) and Rabak (OR: 1.354, 95% CI: 1.188-1.543) localities compared to Al Douiem locality. Anti-HIV/anti-T. palladium (27.70%) and anti-HIV/HBVsAg (23.07%) were the most frequently detected serologic markers of co-infections, P = 0.002.