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This study was conducted to evaluate the antimicrobial and preservative effects of the combinations of nisin (NS), tea polyphenols (TP), rosemary extract (RE), and chitosan (CS) on pasteurized chicken sausage. An orthogonal test revealed that the most effective preservative was a mixture of 0.05% NS plus 0.05% TP plus 0.03% RE plus 0.55% CS (weight by sausage weight). This mixture had antimicrobial and antioxidant effects in pasteurized chicken sausage and extended the shelf life to >30 days at 4°C. The inhibitory effects of NS, TP, RE, and CS were also evaluated against Pseudomonas aeruginosa, lactic acid bacteria (LAB), and Staphylococcus aureus, the dominant spoilage and pathogenic bacteria in pasteurized chicken sausage. NS had the greatest inhibitory effect on LAB and S. aureus, with inhibitory zone diameters of 19.7 and 17.8 mm, respectively. TP had the largest inhibitory effect on P. aeruginosa, with a clear zone diameter of 18.2 mm. These results indicate that the combination of NS, TP, RE, and CS could be used as a natural preservative to efficiently inhibit the growth of microorganisms in pasteurized chicken sausage and improve its safety and shelf life. Antimicrobial mixtures of nisin, tea polyphenols, rosemary, and chitosan were optimized. The preservation effect of the optimal mixture was evaluated in chicken sausage. Inhibitory effects of each preservative were evaluated against three kinds of bacteria. The optimal mixture significantly extended the shelf life of chicken sausage. [ABSTRACT FROM AUTHOR]