Probiotics refer to microorganisms that exhibit a beneficial effect on the animals’ health through intestinal microbial balance. This investigation intends to identify and define the probiotic characteristics of Lactobacillus strains isolated from the digestive systems of ruminants. Bacterial strains were isolated purified and characterized based on morphological and biochemical characteristics. The isolates were identified using the 16S rRNA gene partial sequencing method. Most of the strains exhibited a decrease in the growth by increasing the concentration of the bile salt, NaCl, increasing the temperature over 37oC, and moving toward neutrality and alkalinity of the media. Strain Lactobacillus farraginis MD_A11 revealed the lowest decrease in the growth percentage when subjected to different bile salt concentrations of 5.96, 6.61, 6.85, 7.40, 7.53, and 7.64%, NaCl% concentrations of 1.40, 5.62, 6.48, 7.36, 7.39, 7.41, 7.42, 7.28, and 13.76%, raising the temperature over 37oC being 4.19 & 3.945%, different pH levels as compared to control, and it recorded the lowest medium pH after the third day being 4.20 with titrable acidity of 0.32%. Strain Lactobacillus farraginis MD_A11 could be recommended as a probiotic feed additive for ruminants to improve their growth performance and productivity.