Abstract
This study aimed to evaluate the compensatory effects of amino acid (AA) supplementation on laying hens fed a low-protein diet, focusing on production performance, egg quality, blood immunity, total tract retention, and intestinal health. A total of 180 Hy-Line Brown laying hens (35 weeks old) were randomly allocated to three dietary treatments with 12 replicates (5 hens/replicate): (1) control diet (15.90% crude protein, CON), (2) reduced-protein diet (15.20% crude protein, NG), and (3) reduced-protein diet supplemented with 400 g/t methionine, 400 g/t lysine, and 300 g/t threonine (15.20% crude protein, LAA). Over the 12-week experimental period, production parameters were monitored weekly. The results showed that compared to the NG group, AA supplementation significantly increased laying rates during weeks 1–4 (p < 0.05) and reduced the feed-to-egg ratio during weeks 9–12 (p < 0.05), with a 3.10% decrease in feed conversion ratio (FCR). By week 12, the NG group exhibited lower Haugh unit values than both the CON and LAA groups (p < 0.05), indicating improved egg quality with AA supplementation. Serum analysis revealed elevated immunoglobulin Y levels and upregulated expression of the anti-inflammatory cytokine IL-10 in the LAA group (p < 0.05). Total tract retention of crude protein (CP) and phosphorus (P) was significantly enhanced in the LAA group compared to CON and NG groups (p < 0.05). However, in terms of cecal microbiota, no significant differences were observed among the experimental groups. These findings demonstrate that AA supplementation alleviates the adverse effects of low-protein diets by enhancing nutrient utilization and immune modulation, thereby improving productive performance, egg quality, and total tract retention in laying hens.