Background: To formulate a nanoemulsion (NE) from cold pressed neem oil and to compare its repellant activity with normal cold pressed oil and N,N-Diethyl-meta-toluamide (DEET) as a standard chemical compound and to improve efficacy of repellency against Anopheles stephensi, through NE formulations.Methods: In this study, complete protection time (CPT) and failure time of samples were evaluated on human subjects using the arm in cage assay, and their values were determined against An. stephensi. The droplet size of the NE were measured with dynamic light scattering. Pseudo ternary phase diagrams were constructed consisting of an emulsion system of an active ingredient (neem oil), surfactant (Tween 80), and water. Two formulations were selected from the isotropic regions and showed particle sizes of 12.3 and 93.9 nm in diameter.Results: The results (expressed in minutes as mean±SD) showed that CPTs of neem oil NE in tween 80, and DEET 20% were 240±39 min and 360±24 respectively. Dose values for Neem oil in formulation and for DEET were 77 μg/cm2 and 308 μg/cm2, respectively. Nano neem oil in tween 80 provided significantly longer protection than normal cold pressed neem oil (P<0.01). The prepared NEs showed high kinetic stability for weeks, without phase separation, sedimentation or creaming. Nevertheless, their droplet size increased slightly over time. Compared to non-formulated neem oil, the NE significantly increased the Repellency of the tested species.Conclusions: NE of neem oil significantly increases the CPT and reduces needed dose values. Hence, neem oil NE can be a good alternative to DEET and other chemical repellents.