Aim: Diabetes is considered as one of the serious complications and disorder in recent years. So, the present work was undertaken to investigate the anti-diabetic activity using Eisenia fetida earthworm extract. Materialsa and Methods: The anti-diabetic activity was carried out by using different concentrations of earthworm, Eisenia fetida extract with two different methods, alpha-amylase inhibitory assay and glucose uptake assay by using standard drug Metformin and Metronidazole respectively. Results: Results revealed that the earthworm, Eisenia fetida extract exhibit anti-diabetic activity by inhibiting alpha-amylase activity and also by increasing the glucose uptake in the cells. Thus, earthworm Eisenia fetida extract may serve as an alternative anti-diabetic drug to prevent diabetic complications in near future.