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The molecular information of lesser-known bullfrog, Hoplobatrachus litoralis is restricted from Bangladesh (type locality) and Myanmar. The morphological observation further evidenced the range-extension of this species from India and Myanmar. Here, we collected the H. litoralis specimen from the Dampa Tiger Reserve, Mizoram state and provides a new altitude record (268 m) and range-extension in northeast India. The DNA barcoding with mitochondrial Cytochrome b gene discriminates H. litoralis from other congeners with sufficient genetic distance (13.9% to 27.8%). The Bayesian phylogeny revealed monophyletic clustering of all five Hoplobatrachus species and showed the sister relationship of H. litoralis and H. tigerinus. The generated sequences of H. litoralis from northeast India depicted shallow divergence (1.5%) from the sequences generated from Bangladesh, indicates the possible genetic variability due to the alteration of the ecological niche. We suggested further physiological and molecular study with extensive sampling of this species to confirm their actual range distribution, high-altitude adaptation, and gene flow.