Abstract
(1) Background: Improving feed efficiency and the vitality of the reproductive system in the late stage of the egg-laying period is of great significance for prolonging the egg-laying cycle and improving egg quality. In the present study, a new flavonoid, which was extracted from kudzu leaf, was chosen to investigate its effects on the productive performance and egg quality of late-laying hens. (2) Methods: A total of 360 500-day-old Hy-Line Brown layer hens were randomly divided into a control treatment group (no KLF supplementation), and groups that received 0.2%, 0.4%, 0.6%, 0.8%, and 1.0% KLF supplement treatments. Each treatment contained 6 replicates, with 10 hens in each replicate. Productive performance metrics, including the daily egg production, egg weight, the number of deformed eggs, egg quality, egg density, egg shape index, eggshell strength, yolk color, and the Haugh unit, were meticulously recorded for each replicate. Furthermore, microbial communities and hypothalamus gene expressions were investigated based on the results of the productive performance and egg quality. (3) Results: KLF supplementation significantly decreased the deformity rate while significantly increasing the eggshell strength in the finishing phase afterward (p < 0.05). Specifically, hens supplemented with 0.6% KLF possessed the lowest deformed egg rate. KLF supplementation significantly increased the relative abundances of Bifidobacterium sp., Blautia sp., Lactococcus sp., and Lactobacillus sp., while significantly decreasing Parasutterella sp. and Escherichia-Shigella sp. (p < 0.05). Furthermore, the interactive analysis showed the hypothalamus gene expression mainly interacted with probiotics, such as Bifidobacterium sp. and Lactobacillus sp., through ribosome biogenesis, nucleocytoplasmic transport, and cAMP signaling pathways. (4) Conclusions: The findings of the present study indicate that KLF supplementation significantly proliferated probiotics, such as Bifidobacterium and Lactobacillus, which may have further interacted with hypothalamus genes, thus decreasing the deformity rate while increasing eggshell strength in the finishing phase.