The objective of this research was to develop the nanoliposome-encapsulated phenolic rich fraction from Achillea millefolium (A. millefolium) and to investigate its antibacterial and health-promoting activities in mice challenged by pathogenic foodborne Campylobacter jejuni. The A. millefolium was extracted and the ethyl acetate fraction was found to be the phenolic-rich fraction (PRF) containing 14.72 ± 2.39 mg gallic acid equivalent (GAE)/g dry weight (DM). Base on the results, the synthesized nanoliposome-loaded PRF (PRF-NLs) with the size of 187.2 nm exhibited homogeneous dispersion (PDI 0.213) and moderate stability behavior in colloidal dispersions (Zeta potential −37.45). The non-encapsulated PRF and PRF-NLs were gavaged orally in the mice for 28 days, and mice were challenged with C. jejuni on day 21. The results indicated that the dietary supplementation of non-encapsulated PRF and PRF-NLs significantly (p < 0.05) improved the average daily weight gain, food intake, liver function, antioxidant status, and morphostructural characteristics of the ileum. However, the PRF-NLs appeared to be more potent as compared to non-encapsulated PRF. The higher biological activity of PRF-NLs could be associated with the higher intestinal solubility and absorption of nanoliposome-encapsulated PRF. Thereby, the nanoliposome-encapsulated PRF could be considered as a natural antibiotic alternative called phytobiotic to prevent intestinal infection caused by enteropathogenic C. jejuni.
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