Abstract: In order to study the efficacy of stress Ning in improving the immune and antioxidant properties of commercial broiler chickens. Select 200 healthy 1-day-old AA broiler chickens with similar body weight and randomly divide them into 2 treatments, with 5 replicates per treatment and 20 chickens per replicate (see Table 1). Treatment 1: Control group, broiler chickens were fed a regular diet (NRC recommended nutritional level) without adding any antibiotics to the diet; On the basis of the same diet as treatment 1, supplement stress Ning with 250 mg/kg for treatment 2. The experiment is divided into two stages (0-21d; 22-42d). Experimental chicken cage feeding, feeding pellet feed, with consistent dietary composition and nutritional levels between treatments. At 21 and 42 days, 10 chickens were randomly selected from each treatment (2 chickens/replicate) for slaughter experiments. Blood, liver, and muscle samples were collected to measure inflammatory factors and antioxidant indicators. The measurement results indicate that:

Comparison of cellular inflammatory factors: Compared with the control group, the addition of stress Ning significantly increased the levels of interleukin-6 (IL-6) and chicken tumor necrosis factor at 21 and 42 days (P<0.05), but="" had="" no="" significant="" effect="" on="" other="" indicators="" p="">0.05). From a numerical comparison analysis, with the control group's various indicators as 100%, adding stress Ning increased and decreased the serum chicken tumor necrosis factor - α (IFN - α) by 11.01% in 21 day commercial broiler chickens, increased interleukin-1 β (IL-1 β), and increased interleukin-6 (IL-6) and interleukin-10 (IL-10) by 5.89%, 23.75%, and 10.75%, respectively; Increased the levels of chicken tumor necrosis factor - α (IFN - α), interleukin-1 β (IL-1 β), and interleukin-6 (IL-6) in the serum of 42 day commercial broiler chickens by 20.94%, 5.72%, and 23.10%, respectively, while reducing interleukin-10 (IL-10) by 3.79%.

Antioxidant comparison: Adding stress Ning showed a trend of reducing serum MDA content after 21 days (P=0.063), increasing SOD activity after 21 days (P=0.091), and reducing MDA content after 42 days (P=0.094); Significantly reduced the serum MDA content after 21 days (P<0.05), 21="" 42="" and="" showed="" a="" trend="" of="" increasing="" gsh="" activity="" after="" days="" p="" there="" is="" reducing="" serum="" mda="" levels="" in="" chest="" muscles="" leg="" but="" no="" significant="" effect="" on="" muscle="" sod="">0.05). By numerical comparison and analysis, after adding stress Ning, the serum MDA content, SOD activity, and GSH activity after 21 days were 69.02%, 120.96%, and 108.26%, respectively; The MDA content, SOD activity, and GSH activity in the 42 day serum were 133.45%, 102.59%, and 93.62%, respectively; The MDA content, SOD activity, and GSH activity in the liver after 21 days were 54.76%, 99.44%, and 111.12%, respectively; The serum MDA content, SOD activity, and GSH activity at 42 days were 101.44%, 104.84%, and 112.56%, respectively; The MDA content, SOD activity, and GSH activity in the pectoral muscles were 78.17%, 99.32%, and 89.62%, respectively; The MDA content, SOD activity, and GSH activity of leg muscles were 74.19%, 93.52%, and 106.48%, respectively.

Keywords: commodity broiler, Yanning C1, cytokines, antioxidants

1. Materials and Methods

1.1 Test Materials

Stress Ning: A high-yield stress additive formula mainly composed of microecological active ingredients. Provided by SD BIOTECH GROUP.

1.2 Experimental animals and design

Select 200 healthy 1-day-old AA broiler chickens with similar body weight and randomly divide them into 2 treatments, with 5 replicates per treatment and 20 chickens per replicate (see Table 1). Treatment 1: Control group, broiler chickens were fed a regular diet (NRC recommended nutritional level) without adding any antibiotics to the diet; On the basis of the same diet as treatment 1, supplement stress Ning with 250 mg/kg for treatment 2. The experiment is divided into two stages (0-21d; 22-42d). Feed the control group diet at 28-35 days of age as an endogenous metabolism test to calculate the true metabolic rate of nutrients.

1.3 Experimental diet

Conventional diet for commercial broiler chickens in Shandong Province (see Table 2).

1.4 Test Method

Feeding experiment: The experiment is divided into two stages: broiler chickens (0-21d) and broiler chickens (22-42d). Experimental chicken cage feeding with powdered feed.

Slaughter experiment: 10 chickens were randomly selected for each treatment (2 chickens/replicate) at 21 and 42 days. Conduct slaughter experiments.

1.5 Measurement indicators

Cellular inflammatory factors: At 21 and 42 days, 5 chickens (1 per replicate) were randomly selected from each treatment for blood collection, and the expression of IL-1 β, IL-6, IL-10, and TNF - α was measured.

Blood antioxidant: At 21 and 42 days, 5 chickens were selected for each treatment, blood was collected, serum was separated, and stored at -20 ℃ to determine antioxidant indicators (MDA T-SOD、GSH-PX）。

Organ fluid antioxidant: At the end of the experimental period (slaughter), 5 chickens were selected for each treatment, and liver and chicken samples were taken to measure antioxidant indicators (MDA T-SOD、GSH-PX）。

1.6 Data statistics and analysis

The data was analyzed using SAS 9.2 software, and Duncan's multiple range test was used to compare the mean values, with a significance level of p<0.05.

2. Results and Analysis

2.1 Comparative analysis of serum inflammatory factors

The effect of supplementing 250 mg/kg of stress Ning in the diet of commercial broiler chickens on serum inflammatory factors is shown in Table 3. Analysis of variance showed that compared with the control group, the addition of stress Ning significantly increased the levels of interleukin-6 (IL-6) and chicken tumor necrosis factor at 21 and 42 days (P<0.05), but="" had="" no="" significant="" effect="" on="" other="" indicators="" p="">0.05). From a numerical comparison analysis, with the control group's various indicators as 100%, adding stress Ning increased and decreased the serum chicken tumor necrosis factor - α (IFN - α) by 11.01% in 21 day commercial broiler chickens, increased interleukin-1 β (IL-1 β), and increased interleukin-6 (IL-6) and interleukin-10 (IL-10) by 5.89%, 23.75%, and 10.75%, respectively; Increased the levels of chicken tumor necrosis factor - α (IFN - α), interleukin-1 β (IL-1 β), and interleukin-6 (IL-6) in the serum of 42 day commercial broiler chickens by 20.94%, 5.72%, and 23.10%, respectively, while reducing interleukin-10 (IL-10) by 3.79%. This proves that adding stress Ning to the diet of commercial broiler chickens significantly reduced the levels of chicken tumor necrosis factor - α (IFN - α) in the serum of 21 day broiler chickens compared to the control group, increased the levels of interleukin-1 β (IL-1 β), interleukin-6 (IL-6), and interleukin-10 (IL-10) in the serum of 21 day broiler chickens, and increased the levels of chicken tumor necrosis factor - α (IFN - α), interleukin-1 β (IL-1 β), and interleukin-6 (IL-6) in the serum of 42 day broiler chickens.

2.2 Comparative analysis of serum antioxidant activity

The effect of supplementing 250 mg/kg of stress Ning in the diet of commercial broiler chickens on serum antioxidant performance is shown in Table 4. Analysis of variance showed that the addition of stress Ning had a trend of reducing serum MDA content after 21 days (P=0.063), increasing SOD activity after 21 days (P=0.091), and reducing MDA content after 42 days (P=0.094). By numerical comparison and analysis, after adding stress Ning, the serum MDA content, SOD activity, and GSH activity after 21 days were 69.02%, 120.96%, and 108.26%, respectively; The MDA content, SOD activity, and GSH activity in the 42 day serum were 133.45%, 102.59%, and 93.62%, respectively.

2.3 Comparative analysis of liver antioxidant activity

The effect of supplementing 250 mg/kg of stress Ning in the diet of commercial broiler chickens on liver antioxidant performance is shown in Table 5. Analysis of variance showed that the addition of stress Ning significantly reduced the serum MDA content at 21 days (P<0.05), and showed a trend of increasing GSH activity at 21 days (P=0.086) and 42 days (P=0.050). By numerical comparison and analysis, the MDA content, SOD activity, and GSH activity in the liver after 21 days of adding stress Ning were 54.76%, 99.44%, and 111.12%, respectively; The MDA content, SOD activity, and GSH activity in the 42 day serum were 101.44%, 104.84%, and 112.56%, respectively.

2.4 Comparative analysis of muscle antioxidant activity

The effect of supplementing 250 mg/kg of stress Ning in the diet of commercial broiler chickens on the antioxidant properties of chest and leg muscles is shown in Table 6. Analysis of variance showed that the addition of stress Ning had a trend of reducing serum MDA levels in chest muscles (P=0.076) and leg muscles (P=0.064), but had no significant effect on SOD and GSH activities (P>0.05). Comparing and analyzing numerically, after adding stress Ning, the MDA content, SOD activity, and GSH activity in the chest muscles were 78.17%, 99.32%, and 89.62%, respectively; The MDA content, SOD activity, and GSH activity of leg muscles were 74.19%, 93.52%, and 106.48%, respectively.

3. Conclusion

Compared to the control group:

Adding stress Ning to commercial broiler chickens significantly reduced the levels of chicken tumor necrosis factor - α (IFN - α) in the serum of 21 day broiler chickens, increased the levels of interleukin-1 β (IL-1 β), interleukin-6 (IL-6), and interleukin-10 (IL-10) in the serum of 21 day broiler chickens, and increased the levels of chicken tumor necrosis factor - α (IFN - α), interleukin-1 β (IL-1 β), and interleukin-6 (IL-6) in the serum of 42 day broiler chickens.

Adding stress Ning to commercial broiler chickens improved serum antioxidant performance after 21 days, reducing MDA content by 30.98%, and increasing SOD activity and GSH activity by 20.96% and 8.26%, respectively; Improved the MDA content and GSH activity in the liver at 21 days, as well as the SOD and GSH activities at 42 days, resulting in a 45.24% decrease in MDA content at 21 days, and an 11.12%, 4.84%, and 12.56% increase in GSH activity at 21 days, SOD activity, and GSH activity at 42 days, respectively; Improved the antioxidant index of muscle MDA, reducing the MDA content in chest and leg muscles by 21.83% and 25.81%, respectively.

Table 1 Experimental Design

Table 2 Experimental Diet Composition and Nutritional Level

Table 3: Effects of Stress Ning on Serum Inflammatory Factors in Commercial Broiler Chickens

 Table 4: Effects of Stress Ning on Serum Antioxidants in Commercial Broiler Chickens

Table 5: Effects of Stress Ning on Antioxidant Properties of Commercial Broiler Liver

 Table 6 Effects of Stress Ning on Antioxidant Properties of Chest and Leg Muscles in Commercial Broiler Chickens at 42 Days