Japanese encephalitis virus (JEV) comes under the family Flaviviridae and genus flavivirus. Pigs act as reservoir and amplifying intermediate host for JEV. The current investigation was conducted to understand the prevalence of JEV infection in pigs in three different geographical sites in India (Odisha, Assam and Manipur). Total 857 serum samples were tested by ELISA and RT-PCR, while only RT-PCR was performed in case of 275 tonsils tissues for detection of JEV. It was observed that JEV prevalence was highest in Manipur (positive 39, 25.45% in serum and 10.08% in tonsil) but lower in Assam (positive 15, 3.75% in serum and 0% in tonsils) and Odisha (positive 7, 1.49% in serum and 3.7% in tonsils). Genotype III (GIII) of JEV was the dominant genotype. Further, analysis of E gene revealed sporadic mutations of S83G, H76P, E78Q, C55S, and S64W along with two consistent mutations V46S and V51I in GIII. Whereas, a single mutation S118N was observed in the GI strain. In conclusion, the high JE virus infection rate of pig in the current locations suggests the need for continuous surveillance of this virus in pigs which will ultimately help to adopt an effective control strategy to prevent the spread of JE infection to human.