Background: Hepatitis B virus (HBV) infection is a global public health concern, affecting an estimated 240 million individuals worldwide. Blood donors' knowledge and practices related to HBV remain underexplored. Objective: This study aimed to assess the knowledge and practices of blood donors regarding nutritional management and prevention of HBV. Method: A cross-sectional study was conducted with 120 blood donors using a simple random sampling technique and semi-structured face-to-face interviews. Data were analyzed using statistical procedures such as t-tests, Pearson Chi-square tests, and frequency distributions. Result: The mean age of participants was 34.5 years, with 83% female and 50% aged between 31 and 40 years. Results indicated that 56.7% of blood donors exhibited good practices related to HBV, while only 25.0% demonstrated strong knowledge. Statistical analysis revealed that blood donors from rural areas had a significantly higher level of knowledge (p=0.001), with rural participants scoring 0.80 compared to 0.78 in urban participants. Female donors had a higher mean practice score (0.81) than male donors (0.70) with a p-value of 0.02. Additionally, Muslim donors exhibited significantly better practices compared to their counterparts (p=0.035), with Muslim donors scoring 0.89 versus 0.79 among non-Muslim donors. Conclusions: This study highlights the gaps in knowledge and practices concerning HBV among blood donors, especially regarding nutritional management. Gender, location, and religion significantly influenced knowledge and practice scores.