The present study aimed to discriminate pistachio (Pistacia vera L.) hulls belonging to three different Tunisian geographical origins and extracted separately by hexane, acetone, acetonitrile, and water in terms of phytochemicals and antioxidant and antibacterial activities using multivariate analysis. Significant differences (P<0.05) in the phytochemical content, antioxidant, and antifoodborne bacterial activities were detected among the pistachio hulls populations. Pearson correlation, principal component analysis (PCA), hierarchical cluster analysis (HCA), and heat map were used to distinguish the relationship between the different regions on the basis of the biological activities. It was found that the twelve (4 extracts × 3 geographical sources) pistachio hulls extracts could be classified geographically into four distinct groups. To explore the mode of action of the aqueous pistachio hull extract against L. monocytogenes and S. enterica, polymyxin acriflavine lithium chloride ceftazidime aesculin mannitol (PALCAM) and xylose lysine deoxycholate (XLD) broth media were artificially contaminated at 104 CFU/mL. Using linear and general linear models, aqueous pistachio hull extract was demonstrated to control the two dominant food-borne pathogens by suppressing the bacterial growth.