Purpose: Given the increasing use of antibiotics on humans and livestock for treatment or as a growth stimulant, antibiotic resistance has become a general concern. The food chain and specially fermented foods could be a source of antibiotic-resistant bacteria and resistance genes. Lactic Acid Bacteria (LAB) and Lactobacilli are considered safe to use as starter culture or probiotic strains. Recently, however, antibiotic-resistant genes isolated from LABs showed the necessity of setting international regulations to reduce the risk of antibiotic resistance genes transmission via the food chain. The current study aimed to investigate the antibiotic resistance of Lactobacilli isolated from traditional yogurt samples from Zanjan province in Iran.Methods: Lactobacilli characterization and identification were carried out through biochemical and molecular methods. The disk diffusion method was applied to determine phenotype resistance using 13 antibiotic disks resistance genes presence were investigated in the isolates to determine transferability risk, respectively.Results: Based on biochemical and molecular methods, 24 isolates have been identified as Lactobacilli with multiple antibiotic-resistant phenotypes. Vancomycin resistance was a typical phenotype and genotype among isolates. On investigated Lactobacilli chromosome, Tetracycline resistance genes Chloramphenicol (cat), beta-lactam, aminoglycosides (aph (3’)-III), and aadA resistance genes have been detected. While the examined resistance genes have not been detected on the plasmids, they were all on the bacterial chromosome.Conclusion: The results showed that the investigated isolates did not carry the resistance genes on their plasmids. It, therefore, would be a good point since they probably do not transfer resistance genes to other bacteria, and they would be proper candidates to do more investigation for introducing new safe starter culture or probiotic strain to food industries.