Abstract: This study was designated to investigate the antiviral activity of the extracellular extract of Lactobacillus bulgaricus 761N against Hepatitis C virus. The cytotoxicity of the extracellular extract of L. bulgaricus 761N assay was conducted on two cell lines involving the Human Liver Hepatocellular Carcinoma (HepG2) cell line and the human Peripheral Blood Mononuclear Cells (PBMC). The antiviral capacity of the extracellular extract of the candidate strain against Hepatitis C Virus (HCV) was tested using real time quantitative polymerase chain reaction (RTqPCR). The pre-treatment trial was found to be of a higher a ntiviral activity than the post-treatment trial. It was found that it possesses also a moderate antioxidant activity that was observed to be higher on HepG2 cell line than on PBMC. Purification by the fractionation process using size exclusion chromatography revealed the presence of two fractions (8 and 15) that were confirmed by SDS-PAGE and were of molecular weights 14.4 and 45 kDa, respectively. Fraction 8 was found to be of a higher antiviral bioactivity in case of pretreatment trial than fraction 15. In case of post-treatment trial, fraction 8 was also observed to be of a higher antiviral capacity than fraction 15.