Abstract: The probiotic features and safety of Lactobacillus species from different foods in the advanced countries have been extensively studied. It is yet to be understood if the lactic acid bacteria that are involved in fermentation of some indigenous foods in Africa are probiotic. The probiotic characteristics of five Lactobacillus species isolated from local raw cow milk in Awka metropolis in Nigeria were evaluated by in vitro and in vivo methods. The in vitro tests used were: Tolerance to low pH and bovine bile, cell surface hydrophobicity and antimicrobial activities. Three strains showed good probiotic potentials in vitro and were selected for in vivo studies. Twenty albino rats were weighed and randomized into five groups in accordance with different treatments for each group. Three groups were administered orally with the three strains each (0.5 mL of 108-1010 CFU mL-1 daily), while the two control groups received basal diet and sterilized skimmed milk, respectively, for six days. The blood samples were collected at the end of a ten-day post ingestion period and analysed for some blood biomarkers which included serum alanine aminotransferase, aspartate aminotransferase, serum cholesterol and haemoglobin. Results showed that three test groups had higher mean weights (203.90, 195.18 and 170.77 g) that were significantly different (p<0.05) from one of the control groups (119.77 g). The strains survived the gastrointestinal tract as evidenced by the Lactobacilli viable count (9.45-10.47 log CFU g-1 faeces) in the test groups. The results of the blood biomarkers showed the three strains improved the health status of the albino rats in the test groups, to various degrees, compared to controls. Generally, Lactobacillus AC showed the best probiotic characteristics and could be safely used in probiotic formulations or in co-culture with other starter bacteria of fermented foods.