Abstract
The aim of this study was to evaluate the effects of Flos lonicerae and Baikal skullcap extracts (PE) on laying performance, antioxidant capacity, immune function, follicular development, estrogen secretion, ovarian metabolomics, and cecal microbiota in aged laying hens. The total number of 70-week-old XinYang Black-Feathered laying hens was 240. These hens were randomly divided into two groups, with each group consisting of six replicates of 20 birds. Control (CON) group was fed a basal diet, whereas the PE group received the same basal diet supplemented with 500 mg/kg of PE. The duration of the experiment was 10 weeks. The findings indicated that the supplementation of PE improved laying performance, antioxidant capacity, and immune function. This was reflected by significant increases (p < 0.05) in laying rate, feed conversion ratio, antioxidant indicators (such as glutathione peroxidase, total antioxidant capacity, and catalase), and immunoglobulin levels. Additionally, there were notable decreases (p < 0.05) in the malondialdehyde levels and pro-inflammatory markers. Moreover, the PE group exhibited a greater number of large yellow and white follicles, as well as higher serum estrogen levels, compared to the CON group (p < 0.05). 16S rRNA sequencing revealed that PE supplementation altered the composition of the cecal microbiota by increasing Ruminococcus_torques_group, Butyricoccus and Christensenellaceae_R-7_group abundances and decreasing Bacteroides, Prevotellaceae_UCG-001 and Megamonas abundances (at genus level), which are primarily associated with short-chain fatty acid production. Ovarian metabolomic analysis showed that the major metabolites altered by PE supplementation were mainly involved in follicular development, estrogen biosynthesis, anti-inflammatory and antioxidant properties. Moreover, changes in both the cecal microbiota (at genus level) and ovarian metabolites were strongly correlated with laying performance, antioxidant status, and immune function. In conclusion, PE supplementation improved laying performance in aged hens by enhancing antioxidant, immune, and ovarian functions, promoting follicular development and estrogen secretion, and modulating the gut microbiota and ovarian metabolites. These findings will offer novel insights into the mechanisms that underlie egg production in the ovaries of aged poultry.