Earthworms from various land uses in the Kottayam district of Kerala, part of the Western Ghats hotspot in India, were studied using an integrative approach that involved morphoanatomical methods and cytochrome c oxidase I (COI) barcoding techniques for the first time here. The study unveiled the presence of nine earthworm species: <i>Drawida ghatensis</i> Michaelsen, 1910; <i>Moniligaster julkai</i> Narayanan and Paliwal, 2022; <i>Glyphidrilus annandalei</i> Michaelsen, 1910; <i>Pontoscolex corethrurus</i> (Müller, 1857); <i>Eukerria kuekenthali</i> (Michaelsen, 1908); <i>Dichogaster bolaui</i> (Michaelsen, 1891); <i>Megascolex cochinensis</i> Stephenson, 1915; <i>M. konkanensis</i> Fedarb, 1898; and <i>Lampito mauritii</i> Kinberg, 1867, belonging to eight genera. The COI sequences of all nine species were deposited in GenBank. Notably, <i>M. julkai</i> and <i>E. kuekenthali</i> are novel additions to the GenBank barcode database. The Bayesian phylogram and maximum-likelihood indicated that the species <i>M. julkai</i> in this study exhibited a closer relationship to <i>D. ghatensis</i> than to other <i>Moniligaster</i> species. This study provides detailed descriptions and illustrations of all nine species, supplemented by their DNA barcodes, to facilitate future species identification.