White feather broiler feed containing bunge pricklyash leaves and preparation method of white feather broiler feed

Abstract

The invention discloses a white feather broiler feed containing bunge pricklyash leaves and a preparation method of the white feather broiler feed, and belongs to the technical field of feed preparation. By adjusting the formula proportion of the feed and adding proper mineral substances, the basic nutritional requirements of the white feather broilers are met, the unconventional feed raw material bunge pricklyash leaves are used, no adverse effect is caused on the growth performance of the broilers, the apparent metabolic rate of nutrient substances such as DM, CP and ASH can be increased, and by improving the antioxidant indexes of serum and liver and up-regulating Nrf2 and NQO1 signal channels of Nrf2 / HO-1, the yield of the broilers is increased. Through expression of genes such as HO-1, SOD1 and CAT, the antioxidant capacity of an organism is improved, oxidative stress in the breeding process is effectively relieved, valuable reference is provided for development and utilization of unconventional feed in China, and sustainable development of the broiler chicken breeding industry is promoted.

Description

Technical Field

[0001] The invention belongs to the technical field of feed preparation, and particularly relates to a white-feather broiler feed containing Zanthoxylum bungeanum leaves and a preparation method thereof. Background Art

[0002] Nowadays, feed formula models mainly include corn soybean meal type and wheat soybean meal type. The main raw materials used in poultry feed in my country are corn, soybean meal and other by-products of grain processing, but conventional bulk feed raw materials are far from meeting the needs of animal husbandry development, especially the shortage of feed grains and protein feed, which has become an important bottleneck restricting the sustainable development of animal husbandry. At present, most common feed formulas use various miscellaneous meal (cottonseed meal, rapeseed meal, peanut meal, etc.), grain processing by-products, fermentation industry by-products, starch sugar by-products and other unconventional feed raw materials. Although these raw materials have a huge total amount, a wide range of sources, and huge nutritional potential, they are complex in composition, large in variation, unstable in quality, and some contain anti-nutritional substances or toxins, which require special treatment before use, as well as poor palatability and low feed intake, resulting in low feeding value and other unfavorable factors. Zanthoxylum bungeanum leaves are currently widely used in the field of medicine and are included in many medical books. They contain rich bioactive substances such as flavonoids, polysaccharides, polyphenols, and are also rich in protein, minerals and dietary fiber. They have good biological effects such as antioxidant, anti-tumor, anti-inflammatory, antibacterial and anti-parasitic. In addition, my country's pepper industry is well developed and rich in resources. Its cultivation history can be traced back thousands of years ago. The planting area is extensive and the yield is considerable. However, compared with the widespread application of pepper fruits, the utilization rate of pepper leaves is relatively low. If pepper leaves are used reasonably and efficiently, it will help alleviate the tension of feed resources in my country and provide a natural and healthy feed source for broilers in large-scale farming, thereby reducing stress reactions. Summary of the invention

[0003] The technical problem to be solved by the present invention is to provide a white-feather broiler feed containing Zanthoxylum bungeanum leaves and a preparation method thereof, so as to solve the technical problem that chickens are prone to oxidative stress during the breeding process.

[0004] To achieve the above-mentioned purpose, the technical solution adopted by the present invention is: to provide a white-feather broiler feed containing Zanthoxylum bungeanum leaves, and the raw materials used include, by weight: 55.87-64.64 parts of corn, 27-32.74 parts of soybean meal, 0-4 parts of puffed soybeans, 2-2.5 parts of fish meal, 2-2.5 parts of soybean oil, 1.1-1.4 parts of calcium hydrogen phosphate, 1-1.6 parts of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.24-0.25 parts of L-lysine hydrochloride, 0.2-0.22 parts of DL-methionine, 0.3 parts of premix, 0-2.3 parts of rice bran and 0.1-3 parts of Zanthoxylum bungeanum leaves.

[0005] On the basis of the above technical solution, the present invention can also be improved as follows:

[0006] Furthermore, the raw materials used include, by mass: 60 parts of corn, 30 parts of soybean meal, 2 parts of puffed soybeans, 2.2 parts of fish meal, 2.2 parts of soybean oil, 1.3 parts of calcium hydrogen phosphate, 1.3 parts of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.24 parts of L-lysine hydrochloride, 0.21 parts of DL-methionine, 0.3 parts of premix, 1 part of rice bran and 1.5 parts of Zanthoxylum bungeanum leaves.

[0007] Furthermore, the raw materials used include, by mass, 55.87 parts of corn, 32.74 parts of soybean meal, 4 parts of puffed soybeans, 2.5 parts of fish meal, 2 parts of soybean oil, 1.4 parts of calcium hydrogen phosphate, 1 part of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.25 parts of L-lysine hydrochloride, 0.22 parts of DL-methionine, 0.3 parts of premix and 0.1 parts of Zanthoxylum bungeanum leaves.

[0008] Furthermore, the raw materials used include, by mass, 64.64 parts of corn, 27 parts of soybean meal, 2 parts of fish meal, 2.5 parts of soybean oil, 1.1 parts of calcium hydrogen phosphate, 1.6 parts of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.24 parts of L-lysine hydrochloride, 0.2 parts of DL-methionine, 0.3 parts of premix, 2.3 parts of rice bran and 3 parts of Zanthoxylum bungeanum leaves.

[0009] Furthermore, the premix comprises, per kg, 16 mg copper, 20 mg iron, 1.25 mg iodine, 120 mg manganese, 110 mg zinc, 0.3 mg selenium, 9000-11000 IU vitamin A, 4000-4750 IU vitamin D3, 55-72.5 IU vitamin E, 2.2-3.1 mg vitamin K3, 5.4-7.55 mg vitamin B2, 2.2-2.85 mg vitamin B1, 2.2-2.85 mg vitamin B6, 2.2-3.75 mg vitamin B 12 0.011-0.017mg, pantothenic acid 15-19mg, niacin 45-62.5mg, biotin 0.15-0.2mg, folic acid 1.6-2.05mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

[0010] Furthermore, the premix includes, per kg, 16 mg copper, 20 mg iron, 1.25 mg iodine, 120 mg manganese, 110 mg zinc, 0.3 mg selenium, 11000 IU vitamin A, 4750 IU vitamin D3, 72.5 IU vitamin E, 3.1 mg vitamin K3, 7.55 mg vitamin B2, 2.85 mg vitamin B1, 3.75 mg vitamin B6, and 100 mg vitamin B6. 120.017mg, pantothenic acid 19mg, niacin 62.5mg, biotin 0.2mg, folic acid 2.05mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

[0011] Furthermore, the premix includes, per kg, 16 mg copper, 20 mg iron, 1.25 mg iodine, 120 mg manganese, 110 mg zinc, 0.3 mg selenium, 9000 IU vitamin A, 4000 IU vitamin D3, 55 IU vitamin E, 2.2 mg vitamin K3, 5.4 mg vitamin B2, 2.2 mg vitamin B1, 2.2 mg vitamin B6, 2.2 mg vitamin B 12 0.011mg, pantothenic acid 15mg, niacin 45mg, biotin 0.15mg, folic acid 1.6mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

[0012] The invention also discloses a method for preparing white-feather broiler feed containing Zanthoxylum bungeanum leaves, which comprises the following steps: mixing raw materials in proportion to obtain the white-feather broiler feed containing Zanthoxylum bungeanum leaves.

[0013] The beneficial effects of the present invention are as follows: the present invention meets the basic nutritional requirements of white-feathered broilers by adjusting the formula ratio of the feed and adding appropriate minerals, uses the unconventional feed raw material Zanthoxylum bungeanum leaves, has no adverse effect on the growth performance of broilers, can improve the apparent metabolic rate of nutrients such as DM, CP, ASH, etc., and improves the body's antioxidant capacity by improving serum and liver antioxidant indexes and up-regulating the expression of genes such as Nrf2, NQO1, HO-1, SOD1 and CAT in the Nrf2 / HO-1 signaling pathway, effectively alleviating oxidative stress in the breeding process, providing a valuable reference for the development and utilization of unconventional feeds in my country, and being conducive to promoting the sustainable development of the broiler breeding industry. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 This is the activity diagram of serum antioxidant index T-AOC;

[0015] Figure 2 This is the activity diagram of serum antioxidant index T-SOD;

[0016] Figure 3 This is the activity diagram of serum antioxidant index GSH-PX;

[0017] Figure 4 This is the activity graph of serum antioxidant index CAT;

[0018] Figure 5 This is the MDA content chart of serum antioxidant index;

[0019] Figure 6This is the activity diagram of T-AOC, an antioxidant index of the liver;

[0020] Figure 7 This is the activity diagram of T-SOD, an antioxidant index of the liver;

[0021] Figure 8 This is the activity diagram of GSH-PX, an antioxidant index of the liver;

[0022] Fig. 9 This is the CAT activity graph of the liver antioxidant index;

[0023] Fig.10 This is the MDA level graph of liver antioxidant index;

[0024] Fig.11 This is the relative expression of the antioxidant gene Nrf2 in the jejunal mucosa;

[0025] Fig.12 This is the relative expression of the antioxidant gene NQO1 in the jejunal mucosa;

[0026] Fig.13 This is the relative expression of the antioxidant gene HO-1 in the jejunal mucosa;

[0027] Fig.14 This is the relative expression of the antioxidant gene SOD1 in the jejunal mucosa;

[0028] Fig.15 This is the relative expression of the antioxidant gene CAT in the jejunal mucosa;

[0029] Fig.16 This is the relative expression level of the antioxidant gene GPX1 in the jejunal mucosa. DETAILED DESCRIPTION

[0030] The specific embodiments of the present invention are described below to facilitate the understanding of the present invention by those skilled in the art. If the specific conditions are not specified in the examples, the methods are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturers are not specified in the reagents or instruments used, they are all conventional products that can be purchased commercially. However, it should be clear that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, as long as various changes are within the spirit and scope of the present invention defined and determined by the attached claims, these changes are obvious, and all inventions and creations using the concept of the present invention are protected.

[0031] The choline chloride, mildew inhibitors and antioxidants used in the following examples and experimental examples were purchased from Sichuan Animal Science Group Co., Ltd.

[0032] Example 1

[0033] A white-feather broiler feed containing Zanthoxylum bungeanum leaves. The raw materials used include, by weight, 60 parts of corn, 30 parts of soybean meal, 2 parts of puffed soybeans, 2.2 parts of fish meal, 2.2 parts of soybean oil, 1.3 parts of calcium hydrogen phosphate, 1.3 parts of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.24 parts of L-lysine hydrochloride, 0.21 parts of DL-methionine, 0.3 parts of premix, 1 part of rice bran and 1.5 parts of Zanthoxylum bungeanum leaves.

[0034] The premix contains 1kg of copper, 20mg of iron, 1.25mg of iodine, 120mg of manganese, 110mg of zinc, 0.3mg of selenium, 10000IU of vitamin A, 4500IU of vitamin D3, 60IU of vitamin E, 2.7mg of vitamin K3, 6mg of vitamin B2, 2.5mg of vitamin B1, 3mg of vitamin B6, and 10000IU of vitamin A. 12 0.015mg, pantothenic acid 17mg, niacin 55mg, biotin 0.17mg, folic acid 1.8mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

[0035] The preparation method of the white-feathered broiler feed containing Zanthoxylum bungeanum leaves comprises the following steps: crushing corn, soybean meal, puffed soybeans, rice bran and Zanthoxylum bungeanum leaves, then passing through a 40-mesh sieve, and then mixing all the raw materials in proportion to obtain the white-feathered broiler feed containing Zanthoxylum bungeanum leaves.

[0036] Example 2

[0037] A white-feather broiler feed containing Zanthoxylum bungeanum leaves. The raw materials used include, by weight, 55.87 parts of corn, 32.74 parts of soybean meal, 4 parts of puffed soybeans, 2.5 parts of fish meal, 2 parts of soybean oil, 1.4 parts of calcium hydrogen phosphate, 1 part of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.25 parts of L-lysine hydrochloride, 0.22 parts of DL-methionine, 0.3 parts of premix and 0.1 parts of Zanthoxylum bungeanum leaves.

[0038] The premix contains 1kg of copper, 20mg of iron, 1.25mg of iodine, 120mg of manganese, 110mg of zinc, 0.3mg of selenium, 11000IU of vitamin A, 4750IU of vitamin D3, 72.5IU of vitamin E, 3.1mg of vitamin K3, 7.55mg of vitamin B2, 2.85mg of vitamin B1, 3.75mg of vitamin B6, and 1.3mg of vitamin B6. 12 0.017mg, pantothenic acid 19mg, niacin 62.5mg, biotin 0.2mg, folic acid 2.05mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

[0039] The preparation method of the white-feathered broiler feed containing Zanthoxylum bungeanum leaves comprises the following steps: crushing corn, soybean meal, puffed soybeans, rice bran and Zanthoxylum bungeanum leaves, then passing through a 40-mesh sieve, and then mixing all the raw materials in proportion to obtain the white-feathered broiler feed containing Zanthoxylum bungeanum leaves.

[0040] Example 3

[0041] A white-feather broiler feed containing Zanthoxylum bungeanum leaves. The raw materials used include, by weight, 64.64 parts of corn, 27 parts of soybean meal, 2 parts of fish meal, 2.5 parts of soybean oil, 1.1 parts of calcium hydrogen phosphate, 1.6 parts of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.24 parts of L-lysine hydrochloride, 0.2 parts of DL-methionine, 0.3 parts of premix, 2.3 parts of rice bran and 3 parts of Zanthoxylum bungeanum leaves.

[0042] The premix contains 1kg of copper, 20mg of iron, 1.25mg of iodine, 120mg of manganese, 110mg of zinc, 0.3mg of selenium, 9000IU of vitamin A, 4000IU of vitamin D3, 55IU of vitamin E, 2.2mg of vitamin K3, 5.4mg of vitamin B2, 2.2mg of vitamin B1, 2.2mg of vitamin B6, and 1.5mg of vitamin B6. 12 0.011mg, pantothenic acid 15mg, niacin 45mg, biotin 0.15mg, folic acid 1.6mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

[0043] The preparation method of the white-feathered broiler feed containing Zanthoxylum bungeanum leaves comprises the following steps: crushing corn, soybean meal, puffed soybeans, rice bran and Zanthoxylum bungeanum leaves, then passing through a 40-mesh sieve, and then mixing all the raw materials in proportion to obtain the white-feathered broiler feed containing Zanthoxylum bungeanum leaves.

[0044] Experimental Example 1 Growth performance test

[0045] The experiment was divided into two stages: the early stage (1-21 days) and the late stage (22-42 days). 440 healthy 1-day Arbor Acres (AA) broilers (half male and half female) were randomly divided into 4 groups, with 5 replicates in each group in the early stage and 22 chickens in each replicate; the late stage grouping was the same, with 5 replicates in each group and 20 chickens in each replicate. The experiment was divided into 4 groups: the control group (CON) was fed with a corn-soybean meal-based basal diet, the CZ1 group was fed with a basal diet supplemented with 1% Zanthoxylum leaf powder, the CZ2 group was fed with a basal diet supplemented with 2% Zanthoxylum leaf powder, and the CZ3 group was fed with a basal diet supplemented with 3% Zanthoxylum leaf powder. The feed ingredients of the experimental groups are shown in Table 1.

[0046] Table 1 Raw material composition of experimental group

[0047]

[0048] Among them, the premix used in the above experimental group includes 1 kg:

[0049] Brooding period (1-21 days): copper 16 mg, iron 20 mg, iodine 1.25 mg, manganese 120 mg, zinc 110 mg, selenium 0.3 mg, vitamin A 11000 IU, vitamin D3 4750 IU, vitamin E 72.5 IU, vitamin K3 3.1 mg, vitamin B2 7.55 mg, vitamin B1 2.85 mg, vitamin B6 3.75 mg, vitamin B 12 0.017mg, pantothenic acid 19mg, niacin 62.5mg, biotin 0.2mg, folic acid 2.05mg, choline chloride 250mg, mold inhibitor 50mg and antioxidant 200mg;

[0050] Growing period (22-42 days): Copper 16 mg, Iron 20 mg, Iodine 1.25 mg, Manganese 120 mg, Zinc 110 mg, Selenium 0.3 mg, Vitamin A 9000 IU, Vitamin D3 4000 IU, Vitamin E 55 IU, Vitamin K3 2.2 mg, Vitamin B2 5.4 mg, Vitamin B1 2.2 mg, Vitamin B6 2.2 mg, Vitamin B 12 0.011mg, pantothenic acid 15mg, niacin 45mg, biotin 0.15mg, folic acid 1.6mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

[0051] During the experiment, each chicken was weighed on an empty stomach on the 1st, 21st and 42nd day after the start of the experiment (feed was cut off at 8 pm the day before weighing, and drinking water was normal during this period). The daily feed intake of each cage was recorded during the experiment. At the end of the experiment, the average daily weight gain, average daily feed intake and feed-to-weight ratio of each group of single broilers were calculated with repetitions as the calculation unit. In the first week before the chicks entered, the heating, ventilation and water tank equipment of the chicken house were strictly checked and cleaned and disinfected. In the first week of the experiment, the temperature of the chicken house was maintained at 33-35℃, and then decreased by 3℃ every week. When it dropped to 24℃, it was no longer cooled. The relative humidity was controlled at 65-70% for 1-2 weeks of age and reduced to 60-65% for 3-4 weeks of age. Continuous light was used during the experiment, and the dark period at night was gradually increased with the increase of age. Feed twice a day, at 8:00 and 14:30, and drink water freely. Do a good job in immunization and disease prevention of broiler chickens. At 2 weeks of age, add Newcastle disease vaccine ( Newcastle disease virus live freeze-dried LaSota B1 strain). Observe the growth status, mental state, feces morphology and color of broilers at any time, and take measures such as heat preservation, heatstroke prevention and ventilation in the chicken house. Pay attention to the hygiene of drinking water and the ground, clean the chicken house every morning and evening, and spray the ground with disinfectant. All chickens are kept in cages of the same size (40cm×175cm×155cm), with 22 chickens in each cage in the first stage and 5 chickens in each cage in the second stage until the end of the experiment. The experimental results are shown in Table 2.

[0052] Table 2 Growth performance of broiler chickens

[0053]

[0054]

[0055] As can be seen from the table, compared with the control group (CON), the addition of 3% Zanthoxylum leaves to the diet (CZ3 group) significantly reduced the body weight of broilers at 42 days (P<0.05); while the addition of 1% Zanthoxylum leaves to the diet (CZ1 group) and 2% Zanthoxylum leaves to the diet (CZ2 group) had no significant effect on the body weight of broilers at 21 days and 42 days. In addition, as the addition ratio of Zanthoxylum leaves increased, the body weight of broilers at 21 days (P=0.011) and 42 days (P=0.001) increased twice.

[0056] From 1 to 21 days of the experiment, compared with the control group, the addition of 1% and 2% Zanthoxylum leaves to the diet had a tendency to increase the ADG of broilers (P=0.054) and reduce the F / G of broilers (P=0.081). From 22 to 42 days of the experiment, the addition of 1% and 2% Zanthoxylum leaves to the diet had a tendency to increase the ADFI (P=0.060) and F / G (P=0.053) of broilers, and with the increase of the level of Zanthoxylum leaves in the diet, the F / G of broilers increased linearly (P=0.014); compared with the control group, the addition of 3% Zanthoxylum leaves to the diet significantly reduced the ADG of broilers (P<0.05), and with the increase of the level of Zanthoxylum leaves in the diet, the ADG of broilers increased quadratically (P=0.016). During the experimental period of 1 to 42 days, the addition of 1% and 2% Zanthoxylum bungeanum leaves to the diet tended to increase the ADFI (P=0.084) and F / G (P=0.054) of broilers. At the same time, with the increase of the Zanthoxylum bungeanum leaf level in the diet, the F / G of broilers increased linearly (P=0.008). Compared with the control group, the addition of 3% Zanthoxylum bungeanum leaves to the diet significantly reduced the ADG of broilers (P<0.05). In addition, with the increase of the Zanthoxylum bungeanum leaf level in the diet, the ADG of broilers increased linearly (P=0.003) and quadratically (P=0.001).

[0057] Experimental Example 2 Metabolic Test

[0058] 1 gram of TiO2 was added to each kilogram of diet as an indicator. The contents of dry matter (DM), crude protein (CP), crude ash (ASH), calcium (Ca), and total phosphorus (P) were determined with reference to the Association of Official Analytical Chemists (AOAC, 2012) and national standards. The dry matter was determined by the (103±2℃) drying method (GB / T 6435-2014), the crude protein was determined by the nitrogen analyzer method (GB / T 6432-2018), the crude ash was determined by the 550℃ ashing method (GB / T 6438-2007), the calcium was determined with reference to GB / T6436-2018, and the total phosphorus was determined by spectrophotometry (GB / T 6437-2018); the calculation formula for the apparent metabolic rate of nutrients is:

[0059] Nutrient apparent metabolic rate (%) = 100-[(nutrient content in feces × TiO2 content in feed / nutrient content in feed × TiO2 in feces

[0060] content)×100].

[0061] The experimental results are shown in Table 3.

[0062] Table 3 Effect of adding Zanthoxylum bungeanum leaf powder to diet on apparent metabolic rate of broiler chickens (%)

[0063]

[0064] From the table, we can see that during the 12th to 14th day of the experiment, the apparent metabolic rates of DM, CP, ASH, Ca, and P in broilers with 1% and 2% Zanthoxylum leaf powder added to the diet were significantly higher than those in the 3% addition group (P<0.05), but there was no significant difference compared with the control group. During the 33rd to 35th day of the experiment, the apparent metabolic rate of ASH in broilers with 2% and 3% Zanthoxylum leaf powder added to the diet was significantly lower than that in the control group and the 1% addition group (P<0.05).

[0065] Experimental Example 3 Antioxidant Performance Test

[0066] 1. Determination of Antioxidant Capacity of Serum and Liver

[0067] According to the kit instructions: the total antioxidant capacity of serum and liver tissue was determined by ABTS method, the malondialdehyde content was determined by TBA method, the total superoxide dismutase activity was determined by hydroxylamine method, the glutathione peroxidase activity was determined by dithiodinitrobenzoic acid method, and the catalase activity was determined by ammonium molybdate method. All the kits were purchased from Nanjing Jiancheng Bioengineering Institute and determined according to the kit instructions. The above indicators were all determined using Bio-Tek elx800 microplate reader (Bio-Tek, USA).

[0068] 2. Determination of relative gene expression of antioxidant in jejunal mucosa

[0069] Primers for genes such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), nuclear factor erythroid derived-2 (Nrf2), quinone oxidoreductase 1 (NADPH:QuinoneOxidoreductase 1, NQO1), heme oxygenase-1 (Heme Oxygenase-1, HO-1), superoxide dismutase 1 (Superoxide dismutase 1, SOD1), catalase (CAT), and glutathione peroxidase-1 (Glutathione peroxidase-1, GPX1) in broiler jejunal intestinal mucosal tissue were designed and synthesized by Sangon Biotech (Shanghai) Co., Ltd. The primer sequences are shown in Table 4. The RNA of jejunal mucosa of broiler chickens was extracted using the SteadyPure universal RNA extraction kit (Accurate Biology, AG21022) from Accurate Biotechnology Co., Ltd. (Changsha, China). The concentration and purity of RNA (A260 / 280, A260 / 230) were then determined using a nucleic acid detector. The samples with qualified concentration and purity were reverse transcribed into cDNA using the Evo M-MLV reverse transcription premix kit (Accurate Biology, AG11728). PCR analysis was then performed using the SYBRGreen Pro Taq HS premix qPCR kit (Accurate Biology, AG11701). The total volume of the reaction system was 10 μL; SYBR (2×) 5 μL; 0.2 μL of each upstream and downstream primer (10 μM), 1 μL of sample cDNA, and 3.6 μL of sterile enzyme-free water. Finally, an ABI 7500 real-time fluorescence quantitative PCR instrument was used for amplification, and the thermal cycle parameters were set as follows: the reaction conditions were set as follows: 95°C, 30 seconds, 1 cycle; 95°C, 5 seconds, 60°C, 30 seconds, 40 cycles, two replicates for each sample. The comparative Ct method was used to calculate the relative quantitative expression difference:

[0070] Target gene expression level = 2 -△△Ct ,

[0071] △△Ct=(Ct target gene-Ct reference gene) experimental group-(Ct target gene-Ct reference gene) control group, where 2 -△△Ct It indicates the fold change of the target gene expression in the experimental group relative to that in the control group.

[0072] Table 4 PCR primer gene sequence list

[0073]

[0074]

[0075] The experimental results are as follows Figure 1-Figure 11 As shown in the figure, A represents the P value of one-way analysis of variance, L represents the P value of linear analysis, and Q represents the P value of quadratic analysis.

[0076] from Figure 1-Figure 5 It can be seen that the addition of 1% and 2% Zanthoxylum leaves to the diet made the CAT activity in the serum of broilers on the 21st day and the T-SOD and CAT activities in the serum of broilers on the 42th day higher than those in the control group and the 3% addition group (P<0.05). Compared with the control group, the addition of 3% Zanthoxylum leaves to the diet significantly reduced the CAT activity in the serum of broilers on the 42th day (P<0.05). On the 21st day of the experiment, the addition of 2% Zanthoxylum leaves to the diet resulted in significantly higher GSH-PX activity in the serum of broilers than in the control group and the 3% addition group (P<0.05); compared with the control group, the addition of 2% Zanthoxylum leaves to the diet could significantly reduce the MDA content in the serum of broilers (P<0.05). In addition, the addition of Zanthoxylum leaves to the diet had no significant effect on the T-AOC, GSH-PX activity and MDA content in the serum of broilers on the 42th day (P>0.05).

[0077] from Figure 6-Figure 10 It can be seen that the addition of Zanthoxylum bungeanum leaves to the diet had no significant effect on the GSH-PX and CAT activities in the liver of broilers at 21 days and 42 days (P>0.05). On the 21st day of the experiment, the addition of 1% Zanthoxylum bungeanum leaves to the diet made the T-AOC activity in the liver of broilers significantly higher than that in the control group and the 3% addition group; compared with the 3% addition group, the addition of 1% and 2% Zanthoxylum bungeanum leaves to the diet significantly reduced the MDA content in the liver of broilers (P<0.05). On the 42nd day of the experiment, compared with the 3% addition group, the addition of 1% Zanthoxylum bungeanum leaves to the diet significantly increased the T-AOC and T-SOD activities in the liver of broilers (P<0.05).

[0078] from Figure 11-Figure 16It can be seen that there was no difference in the relative mRNA expression of GPX1 gene in the jejunal mucosa of broilers at 21d and 42d, and NQO1 gene in the jejunal mucosa of 42d in each experimental group (P>0.05). Compared with the control group, the addition of 1% and 2% Zanthoxylum leaves to the diet increased the relative mRNA expression of Nrf2, NQO1, HO-1, SOD1 and CAT genes in the jejunal mucosa of broilers at 21d, and they were significantly higher than those in the 3% addition group (P<0.05). Compared with the CZ3 group, the mRNA expression of SOD1 and CAT genes in the jejunal mucosa of broilers at 42d was significantly increased by feeding the diets supplemented with 1% and 2% Zanthoxylum leaves (P<0.05). In addition, the addition of 1% Zanthoxylum leaves to the diet significantly increased the expression of Nrf2 and HO-1 genes in the jejunal mucosa of broilers at 42d than that in the 3% addition group (P<0.05).

Claims

1. A white-feather broiler feed containing Zanthoxylum bungeanum leaves, characterized in that: The raw materials used include, by weight: 55.87-64.64 parts of corn, 27-32.74 parts of soybean meal, 0-4 parts of puffed soybeans, 2-2.5 parts of fish meal, 2-2.5 parts of soybean oil, 1.1-1.4 parts of calcium hydrogen phosphate, 1-1.6 parts of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.24-0.25 parts of L-lysine hydrochloride, 0.2-0.22 parts of DL-methionine, 0.3 parts of premix, 0-2.3 parts of rice bran and 0.1-3 parts of Zanthoxylum bungeanum leaves.

2. The white-feather broiler feed containing Zanthoxylum bungeanum leaves according to claim 1, characterized in that: The raw materials used include, by mass, 60 parts of corn, 30 parts of soybean meal, 2 parts of puffed soybeans, 2.2 parts of fish meal, 2.2 parts of soybean oil, 1.3 parts of calcium hydrogen phosphate, 1.3 parts of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.24 parts of L-lysine hydrochloride, 0.21 parts of DL-methionine, 0.3 parts of premix, 1 part of rice bran and 1.5 parts of Zanthoxylum bungeanum leaves.

3. The white-feather broiler feed containing Zanthoxylum bungeanum leaves according to claim 1, characterized in that: The raw materials used include, by mass, 55.87 parts of corn, 32.74 parts of soybean meal, 4 parts of puffed soybeans, 2.5 parts of fish meal, 2 parts of soybean oil, 1.4 parts of calcium hydrogen phosphate, 1 part of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.25 parts of L-lysine hydrochloride, 0.22 parts of DL-methionine, 0.3 parts of premix and 0.1 parts of Zanthoxylum bungeanum leaves.

4. The white-feather broiler feed containing Zanthoxylum bungeanum leaves according to claim 1, characterized in that: The raw materials used include, by mass, 64.64 parts of corn, 27 parts of soybean meal, 2 parts of fish meal, 2.5 parts of soybean oil, 1.1 parts of calcium hydrogen phosphate, 1.6 parts of stone powder, 0.22 parts of sodium chloride, 0.2 parts of baking soda, 0.24 parts of L-lysine hydrochloride, 0.2 parts of DL-methionine, 0.3 parts of premix, 2.3 parts of rice bran and 3 parts of Zanthoxylum bungeanum leaves.

5. The white-feather broiler feed containing Zanthoxylum bungeanum leaves according to any one of claims 1 to 4, characterized in that: The premix comprises, per kg, 16 mg copper, 20 mg iron, 1.25 mg iodine, 120 mg manganese, 110 mg zinc, 0.3 mg selenium, 9000-11000 IU vitamin A, 4000-4750 IU vitamin D3, 55-72.5 IU vitamin E, 2.2-3.1 mg vitamin K3, 5.4-7.55 mg vitamin B2, 2.2-2.85 mg vitamin B1, 2.2-3.75 mg vitamin B6, 2.2-3.75 mg vitamin B 12 0.011-0.017mg, pantothenic acid 15-19mg, niacin 45-62.5mg, biotin 0.15-0.2mg, folic acid 1.6-2.05mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

6. The white-feather broiler feed containing Zanthoxylum bungeanum leaves according to claim 5, characterized in that: The premix contains 16 mg copper, 20 mg iron, 1.25 mg iodine, 120 mg manganese, 110 mg zinc, 0.3 mg selenium, 11000 IU vitamin A, 4750 IU vitamin D3, 72.5 IU vitamin E, 3.1 mg vitamin K3, 7.55 mg vitamin B2, 2.85 mg vitamin B1, 3.75 mg vitamin B6, and 100 mg vitamin B6. 12 0.017mg, pantothenic acid 19mg, niacin 62.5mg, biotin 0.2mg, folic acid 2.05mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

7. The white-feather broiler feed containing Zanthoxylum bungeanum leaves according to claim 5, characterized in that: The premix contains 16 mg copper, 20 mg iron, 1.25 mg iodine, 120 mg manganese, 110 mg zinc, 0.3 mg selenium, 9000 IU vitamin A, 4000 IU vitamin D3, 55 IU vitamin E, 2.2 mg vitamin K3, 5.4 mg vitamin B2, 2.2 mg vitamin B1, 2.2 mg vitamin B6, 2.2 mg vitamin B 12 0.011mg, pantothenic acid 15mg, niacin 45mg, biotin 0.15mg, folic acid 1.6mg, choline chloride 250mg, antifungal agent 50mg and antioxidant 200mg.

8. The method for preparing the white-feathered broiler feed containing Zanthoxylum bungeanum leaves according to any one of claims 1 to 7, characterized in that: The method comprises the following steps: mixing the raw materials in proportion to obtain white-feather broiler feed containing Zanthoxylum bungeanum leaves.

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