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The present study was carried out to investigate the effects of methionine (Met), lysine (Lys), isoleucine (Ile), and threonine (Thr) deficiency in a low-protein diet on laying performance, egg quality, serum biochemical indices, and the gut microbiota in laying hens. A total of 300 Peking Pink laying hens, at 38 weeks of age, were randomly allocated to five dietary treatments, each of which included six replicates of ten hens. Hens were fed an amino acid-balanced diet (Met: 0.46%; Lys: 0.76%; Ile: 0.72%; Thr: 0.56%; positive control, PC), Met deficiency diet (Met-, 0.25%), Lys deficiency diet (Lys-, 0.56%), Ile deficiency diet (Ile-, 0.54%), and Thr deficiency diet (Thr-, 0.46%) for 12 weeks. Hens were housed in pairs in 45 × 45 × 45 cm wire cages with three ladders and three birds per cage. Feed and water were provided ad libitum during the entire experimental period. All data were analyzed using one-way ANOVA with Turkey's multiple range test. Here, compared to the PC group, final body weight (FBW), average daily gain (ADG), egg production (EP), egg weight (EW), average daily egg mass (EM), average daily feed intake (ADFI), and yield of abdominal fat (AFY) in the Met-group were lower, while EW and EM were higher in the Lys-group. The feed egg ratio (FER) was increased in the Met- and Lys-groups, and EW and AFY were decreased in the Ile-group compared to the controls. Meanwhile, ADG, EP, EM, and ADFI were lower in the Thr group than the PC group. The level of triglycerides (TGs) in the four groups was lower and the concentrations of uric acid (UA) in the Met-group were higher than those in the PC group. The shell color in the Thr group was lower than the PC group. Of note, amino acid deficiency altered the gut microbial structure (e.g., increasing the level of Parabacteroides and decreasing the abundance of Lactobacillus ) in hens. The correlation analysis showed that amino acid deficiency-induced gut microbiota alteration is closely associated with the change in key parameters: FER, UA, AFY, EW, EM, TG, FBW, EP, and ADFI. Collectively, our results highlight the role of adequate amino acid ratio supplementation in the low-crude-protein diet structure for laying hens.

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Alterations and Correlations of the Gut Microbiome, Performance, Egg Quality, and Serum Biochemical Indexes in Laying Hens with Low-Protein Amino Acid-Deficient Diets