The effective group of food supplements known as paraprobiotics offer greater health advantages than conventional nutritional goods, therefore they can be considered functional foods. The traditional buttermilk "Seet," which is made from milk sourced from nearby Hisar, Haryana, communities, was used to collect paraprobiotic bacteria. Following observation of growth, bacteria were isolated on MRS agar media, and pure culture was produced by sub-culturing on the same medium. By morphological analysis, Gram staining, and additional identification by particular biochemical tests, the purity of each culture was established. All the bacterial strains were Gram-positive. Twenty four bacterial isolates were characterized morphologically, culturally, and biochemically, and 13 of them were found to be Lactobacillus spp. The results of the isolates tests for catalase, oxidase, urease, nitrate reduction, Simmon’s citrate, MRVP, Oxidative and Fermentative and Caesin Hydrolysis. All the strains exhibited negative test for catalase, oxidase, urease, nitrate reduction and Simmon’s citrate, but showed positive fermentative and casein hydrolysis test. Maximum samples positively responded to MR test. Few samples (S5, S10, S11, S15, S16 and S24 exhibited VP positive test. A total of 11 strains belonging to 5 genera (Lactobacillus, Pediococcus, Lactococcus, Weissella and Leuconostoc) were identified.