Dietary alfalfa fiber (AF) is conceived to modulate gut microbial richness and diversity to improve the health and growth of weaning piglets. The objective of this study was to evaluate the prebiotic effects of AF on diarrhea incidence, the production of short-chain fatty acids (SCFAs), and microbiota composition in weaning piglets. This study utilized 100 crossbred piglets (Duroc × Landrace × Yorkshire) with a body weight of 8.42 ± 1.88 kg randomly assigned to the following treatments: 0.00% AF meal (A), 6.00% of AF meal (B), 12.00% AF meal (C), and 18.00% AF meal (D). The cecum samples were used to determine microbial community composition and diversity through high-throughput 16S rDNA sequencing. The results of this study show that the lowest average daily gain (ADG) was observed in treatment D, and the highest ADG was recorded in treatment C. However there was no significant difference between the treatment groups and the control. The average daily feed intake (ADFI) was significantly higher in treatment C compared to the other treatments. The feed conversion ratio was high in the control group compared to the AF treated groups. The highest diarrhea incidence was observed in treatment A and the lowest diarrhea incidence was observed in treatment C and D. The highest acetate and propionate levels were observed in treatment B, but there was no significant difference between the treatment groups and the control. The supplementation of AF significantly increased the butyrate level in treatment D compared with treatments A and B but was not significantly different from treatment C. The Observed_species richness and Simpson diversity values of the cecum bacterial composition in the AF fed piglets were higher than the control. In addition, the Chao 1 richness and Shannon diversity increased with an increase in AF supplementation, reaching a plateau at treatment B and C, then decreasing at treatment D. The Bacteroidetes, Firmicutes, Tenericutes, Proteobacteria, Cyanobacteria, Spirochaetae, Actinobacteria, Fibrobacteres, Saccharibacteria, Synergistetes, Chlamydiae, Elusimicrobia, Deferribacteres, Fusobacteria, and others were relatively abundant in all treatments. The Bacteroidetes and Firmicutes were the dominant phyla, accounting for 98% of all reads. AF treatment decreased the Bacteroidetes phylum and increased the Firmicutes phylum compared with treatment A. Therefore, the dietary inclusion of AF may decrease diarrhea incidence, increase cecal bacterial composition and richness, and consequently improve the growth performance of weaning piglets.

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