The present study was conducted to identify a suitable culture medium for the laboratory culture of Drosophila eugracilis. The study evaluated the effectiveness of three different culture media—wheat flour agar, corn flour agar, and banana agar—for the laboratory culture of D. eugracilis by analysing fecundity and productivity. Wild flies were collected from Karnatak University Campus, Dharwad, and cultured under laboratory conditions. The results showed that wheat flour agar medium recorded significantly higher numbers of eggs, pupae, and adult flies compared to corn flour agar and banana agar media. Although banana agar medium contained higher carbohydrate content and protein-to-carbohydrate ratio, it showed lower productivity and greater pupal mortality, likely due to excess sugar content and rapid fungal contamination. No significant difference was observed in developmental duration among the three media. Overall, wheat flour agar medium was found to be the most suitable medium for the successful culture and maintenance of D. eugracilis under laboratory conditions. The study provides preliminary information for establishing stable laboratory cultures of this species for future biological and genetic studies.