This study explores the novel use of mixed cultures of microalgae-Spirulina platensis, Micractinium, and Chlorella-for nutrient removal from dairy wastewater (DW). Microalgae were isolated from a local wastewater treatment plant and cultivated under various light conditions. The results showed significant biomass production, with mixed cultures achieving the highest biomass (2.51 g/L), followed by Spirulina (1.98 g/L) and Chlorella (1.92 g/L). Supplementing DW (75%) with BG medium (25%) significantly enhanced biomass and pH levels, improving pathogenic bacteria removal. Spirulina and mixed cultures exhibited high nitrogen removal efficiencies of 92.56% and 93.34%, respectively, while Chlorella achieved 86.85% nitrogen and 83.45% phosphorus removal. Although growth rates were lower under phosphorus-limited conditions, the microalgae adapted well to real DW, which is essential for effective algal harvesting. Phosphorus removal efficiencies ranged from 69.56% to 86.67%, with mixed cultures achieving the highest removal. Microbial and coliform removal efficiencies reached 97.81%, with elevated pH levels contributing to significant reductions in fecal E. coli and coliform levels. These findings suggest that integrating microalgae cultivation into DW treatment systems can significantly enhance nutrient and pathogen removal, providing a sustainable solution for wastewater management.