Abstract: Background and Objective: The prevalence of liver cancer in Egypt is particularly worrisome. Aflatoxin B1 (AFB1), a hepatocarcinogenic mycotoxin, is one of the main contributors to the high rate of hepatocellular carcinoma (HCC). This study aimed to determine the ability of four lactic acid bacteria (LAB) species in removing or binding aflatoxin B1 (AFB1) from broth media and whole milk and to evaluate the stability of LAB/AFB1 complex during cold storage at 4°C for 1 week. Materials and Methods: Whole milk and MRS broth media were spiked with 50 ng mL–1 AFB1 and incubated at 37°C for 24 h with four LAB strains followed by storage at 4°C for 24, 72 and 196 h. AFB1 removal was determined using HPLC. Results: The efficiency of LAB strains in removing AFB1 from both media was affected by the type of used strain and media. Whole milk was a favorable media for the tested LAB strain in AFB1 reduction when compared with MRS broth. Lactobacillus acidophilus (L. acidophilus) achieved 80% reduction in milk within 24 h at 37°C, whereas Lactobacillus plantarum (L. plantarum) favored cold storage to reduce 85% of the AFB1 content after 1 week. The count of LAB cells in whole milk medium raised in MRS broth by 0.5-2.25 log cycles. None of the tested strains affected by AFB1 (50 ng mL–1) in either MRS or whole milk medium. Conclusion: LAB bacteria were an excellent agent used in minimizing AFB1 in milk at both 37 and 4°C.