Aim: This study aimed to assess and compare the bacterial and viral pathogen loads in wastewater before and after undergoing treatment processes, evaluating the effectiveness of the treatment in pathogen reduction. Materials and Methods: A total of 100 wastewater samples were collected from a wastewater treatment plant, with 50 samples taken from raw influent wastewater and 50 from the effluent after treatment. The samples were analyzed for bacterial pathogens (total coliforms, fecal coliforms, E. coli, Salmonella spp., Shigella spp., and Vibrio cholerae) using standard microbiological methods and viral pathogens (rotavirus, adenovirus, and norovirus) using real-time PCR. Data were analyzed using SPSS, with statistical significance assessed using paired t-tests or Wilcoxon signed-rank tests. Results: The total coliform count reduced by 99.9%, fecal coliforms by 99.75%, and E. coli by 99.78%. Complete removal of Shigella spp. and Vibrio cholerae was observed, with a significant reduction in Salmonella spp. (91.67%). Viral pathogen loads decreased by 99.95% for rotavirus, 97.33% for adenovirus, and over 99.5% for both Genogroups I and II of norovirus, with all reductions being statistically significant (p < 0.001). Conclusion: This study demonstrates that wastewater treatment processes are highly effective in reducing bacterial and viral pathogen loads, providing significant public health benefits. The results support the potential for safely reusing treated wastewater, especially in non-potable applications, contributing to water sustainability and environmental protection.