A total number of 54 adult dairy buffalos were divided into 3 groups based on California Mastitis Test and Somatic Cell Count; group-1 (38 buffalos) with negative CMT and SCC; group-2 (12 buffalos) with positive CMT and SCC and group-3 (4 buffalos) with positive CMT and negative SCC. Ultrasonography of the mammary gland and supramammary lymph nodes was achieved via 7.5 MHz linear transducer. Subclinical mastitis was characterized ultrasonographically by homogenous hypoechoic parenchyma, hypoechogenic contents of gland cistern, the irregular contour lining of teat canal, slightly thickened teat wall, loss the characteristic threelayered appearance of the teat wall and overlapped ill distinct rosette of Furstenberg, papillary ducts and orifice. Superficial inguinal lymph node was enlarged, easily identified and completely hypoechogenic entire structure. Group 2 suffered from subclinical mastitis revealed significant (P≤0.05) increase in lymph node length, depth, elapse area, elapse volume and teat wall thickness, significant (P≤0.05) decrease in gray scale analysis of lymph node and significant (P≤0.05) increase in gray scale analysis of udder parenchyma and cisterns milk contents than that in group 1 and group 3. In conclusion, ultrasonography provides an accurate, non-invasive and rapid field technique for diagnosis of subclinical mastitis in buffalo.