In recent years, interest in probiotics, particularly Lactobacillus strains like Lactobacillus sp. SKG34, isolated from Sumbawa mare milk, has surged due to their potential health benefits. This strain holds promise as a probiotic agent, but understanding its behavior in the gastrointestinal (GI) tract is essential for effective application. This study aimed to investigate the resistance and persistence of Lactobacillus sp. SKG34 in the GI tract of rats. Rats were orally administered the strain at a concentration of 10^8 cells for 30 days. The probiotic’s ability to modulate gut microbiota and improve host health is a key area of interest. By utilizing techniques like PCR-RAPD, this research sought to elucidate the fate of Lactobacillus sp. SKG34 in the complex environment of the rat GI tract. Results revealed a significant increase in LAB population in cecal contents, from 3.06 x 10^9 cfu/g in the control group to 1.25 x 10^10 cfu/g in the probiotic group. RAPD analysis also detected Lactobacillus sp. SKG34 in approximately 23% of the total population of lactic acid bacteria in cecal contents, indicating its resistance and proliferation within the GI tract. Further investigation is warranted to fully understand the potential of Lactobacillus sp. SKG34 as a beneficial probiotic strain for human application.