Available research on non-antibiotic alternative growth promoter (NAGPs) mainly focuses on the effect of a single preparation on the substitution effect of antibiotics. However, different NAGP might work together in harmony. Studies related to NAGP combinations are lacking. This study aimed to screen six non-antibiotic alternative growth promoter combinations (NAGPCs) in vitro so as to find a better alternative method to ensure feed efficiency and the broiler health. Pepsin and 42-d broiler ileal fluid were used for the in vitro digestion test, and the optimal addition levels of NAGPs and NAGPCs were selected based on dry matter (DM) and organic matter (OM) digestibility, microbiota of digestive fluid, and antioxidant capacity of digestive fluid. Results showed the optimum addition amount of Mannose oligosaccharide (MOS), fruit oligosaccharide (FOS), mannanase (MAN), Bacillus subtilis (BS), sodium butyrate (SB), guanidinoacetic acid (CA), xylanase (XYL), glucose oxidase (GO), and phytase (PT) were 0.2%, 0.9%, 0.03%, 0.05%, 0.08%, 0.002%, 0.008%, and 0.004%. The six best combinations were 0.2%MOS + 0.03%MAN + 0.15%SB, 0.2%MOS + 0.03%MAN + 0.05%BS, 0.2%MOS + 0.9%FOS + 0.15%SB, 0.2%MOS + 0.9%FOS + 0.05%BS, 0.2%MOS + 0.9%FOS + 0.03%MAN, and 0.2%MOS + 0.05%BS + 0.004%PT. In conclusion, NAGPCs improved the apparent digestibility of ileal DM and OM, intestinal flora and antioxidant capacity. Comparatively, under this condition, using NAGPC has more beneficial effects than using non-antibiotic alternative growth promoter.
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