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Diatom assemblages were investigated and compared from highlands of Peer Panjal ranges of western Himalayas, India. Samples were collected from surface sediments and stones in 15 sites located in 3 different locations during 2019. A total of 40 taxa (78 species) were reported in the study. Multivariate analyses of diatoms assemblages and physico-chemical variables showed significant difference of species composition within each site, which suggests that regional and climatic factors plays important role in species distribution. Abundance data and common occurrence of diatoms species showed that species like Achnanthidium, Amphora, Encyonema, Discostella, Gomphonema, Halamphora, Hantzschia, Odontidium, Pinnularia and Planothidium were most abundant while some species were seen restricted to particular locations. Conductivity, pH and temperature were the most significant environment variables in the study. The results of our study also highlight the homogeneous communities in these highlands suggesting dominance of pioneer species. Future taxonomic insights based on ecological characterization and comparison would be helpful for predictions for environmental change due to human interference and climate change.