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Drosophila (L.) (Diptera: Drosophilidae) has richly contributed to the understanding of patterns of inheritance, variation, speciation, and evolution. Drosophila, with its cosmopolitan nature and complexities in species compositions, is an excellent model for studying the eco-distributional patterns of various species. This study analyzed the altitudinal and seasonal variation in Drosophila species of Mount Japfu in Nagaland, a sub-Himalayan hilly state of northeast India, over the course of one year. A total of 4,680 Drosophila flies belonging to 19 species of 4 subgenera were collected at altitudes of 1500, 1800, 2100, 2400, and 2700 m a.s.l. The subgenus Sophophora Sturtevant was predominant, with 10 species, followed by subgenus Drosophila, with 4 species. Subgenus Dorsilopha and subgenus Scaptodrosophila were represented by 1 species each. The remaining 3 species were not identified. Cluster analysis and constancy methods were used to analyze the species occurrence qualitatively. Altitudinal changes in the population densities and relative abundances of the different species at different seasons were also studied. The diversity of the Drosophila community was assessed by applying Simpson's diversity index. At 1800 m a.s.l., the Simpson's index was low (0.09301), suggesting high Drosophila diversity at this altitude. The density of Drosophila changed significantly during different seasons (F = 26.72; df = 2; p < 0.0001). The results suggest the distributional pattern of a species or related group of species was uneven in space and time.