Milk is considered as one of the most successful probiotic carriers and bovine (cow) milk is the most predominant food carrier in delivering probiotics. In recent years, there is an increasing demand for probiotic enriched non-bovine milk products such as goat, sheep, camel, and buffalo milk due to their unique compositional and quality characteristics. Although the carrier food matrices have a significant influence on probiotic functionality during processing, storage and gastrointestinal transit, a thorough comparison of carrier milk types on probiotic functional properties is limited. The main objective of this project is to evaluate the probiotic survival and bioaccessibility during gastrointestinal transit and colonic fermentation in the presence of these bovine and non-bovine milk. Cow milk (both full cream and skim), goat milk, and camel milk were inoculated with Lactobacillus acidophilus LA-5 and incubated at 37 °C for 10 hours, samples were stored at 4 °C for 3 weeks. Simulated gastrointestinal digestion and colonic fermentation were performed using fresh and samples stored for 3 weeks. All milk samples exhibited highly satisfactory level of initial probiotic viability (>9.0 log CFU/ml) and a decrease of viability was observed during storage, however, all milk samples maintained above the minimum therapeutic level (>6.0 log CFU/ml) at the end of the storage. The highest probiotic viability during the whole storage period was reported in goat milk. Cow full cream milk and goat milk showed better tolerance to simulated gastrointestinal digestion, and all milk types provided an acceptable gastrointestinal tolerance. No significant (p>0.05) difference was observed between milk types during simulated colonic fermentation, however freshly prepared milk samples demonstrated a better bioaccessibility with a higher viability of ~9.0 log CFU/ml among all milk samples. Hence, intake of probiotic enriched milk product both with bovine and non-bovine origin can result in higher growth, bioaccessibility and potential colonization of probiotics in the host gastrointestinal tract.