Use of ornithine aspartate as a feed additive in animal feed

Abstract

The present invention relates to the use of ornithine aspartate as a feed additive in animal feed, for improving feed absorption and serum biochemical indicators, increasing serum antioxidant capacity, and improving intestinal morphology and structure. Ornithine aspartate is used as a feed additive at a single dose of 300-1500 mg / kg of feed. The present invention adds ornithine aspartate as a feed additive to a daily diet to feed yellow-feathered broiler chickens. The broiler chickens are divided according to their age, and feeds of different properties and components are fed according to their age. By adding ornithine aspartate to the feed, the growth performance of the broiler chickens can be improved, serum biochemical indicators and serum antioxidant capacity can be improved, and the intestinal morphology and structure can be improved.

Description

Technical Field

[0001] The present invention relates to the technical field of animal breeding, in particular to application of ornithine aspartate as a feed additive in animal feed. Background Art

[0002] Broilers are the most commonly raised poultry. Descending from wild jungle fowl, domestication of the domesticated chicken dates back at least 4,000 years, but it wasn't until around 1800 that chicken and eggs became mass-produced commodities. Broilers are typically raised for 42-56 days, reaching market weights of 2.5-3.5 kg. Broilers are known for their delicious meat, rich in protein, vitamins, minerals, and other nutrients, making them highly sought after by consumers. In China, the broiler industry has become a vital component of the livestock sector, providing a rich source of food for the population.

[0003] In broiler farming, muscle energy supply is crucial for fast-growing chickens. Improving intestinal morphology or maintaining intestinal homeostasis is a promising approach to ensure rapid growth. Ornithine aspartate, chemically known as (S)-2,5-diaminopentanoic acid-(S)-2-aminosuccinate, breaks down into two endogenous amino acids, L-aspartate and L-ornithine, upon entry into the body. L-ornithine primarily participates in the urea synthesis cycle, promoting blood ammonia metabolism and ornithine regeneration. Ornithine is also decarboxylated by ornithine decarboxylase to produce putrescine, which is further synthesized into spermidine and spermine, collectively known as polyamines. Polyamines have antioxidant properties and can maintain intestinal homeostasis. L-aspartate can be converted to glutamate by glutamine synthetase, which then combines with highly toxic blood ammonia to form glutamine, thereby lowering blood ammonia levels. Currently, ornithine aspartate is primarily used to treat liver cirrhosis and fatty liver disease, and it also promotes the repair and regeneration of damaged liver cells and restores liver cell function. However, there are few reports on the application of ornithine aspartate in livestock and poultry production. Summary of the Invention

[0004] The purpose of the present invention is to overcome the shortcomings of the existing technology and meet actual needs, and to provide the use of ornithine aspartate as a feed additive in animal feed to improve feed absorption and serum biochemical indicators, enhance serum antioxidant capacity, and improve intestinal morphology and structure.

[0005] In order to achieve the purpose of the present invention, the technical solution adopted by the present invention is:

[0006] The use of ornithine as a feed additive in animal feed is used to improve feed absorption and serum biochemical indicators, increase serum antioxidant capacity, and improve intestinal morphology and structure;

[0007] The feed additive is used in a single dosage of 300-1500 mg / kg feed.

[0008] Preferably, ornithine aspartate is added as a feed additive to the diet of yellow-feathered broiler chickens, wherein the broiler chickens are divided according to their age, and feeds with different properties and components are fed according to their age. During the experiment, the broiler chickens are allowed to eat and drink water freely.

[0009] Wherein, the feeds with different properties and components fed to pigs at different ages are:

[0010] (1) Feeding male yellow-feathered broiler chickens aged 1 to 21 days with a crushed pelleted diet comprising a basic diet A and a feed additive;

[0011] (2) At 22-42 days of age, male yellow-feathered broiler chickens were fed a diet comprising a basic diet B and feed additives.

[0012] Preferably, the mass percentage composition of the basic diet A is: 50-55% corn, 30-35% soybean meal, 4.0-6.0% wheat bran, 0.5-2.0% corn gluten meal, 0.5-2.0% soybean oil, and 4.0-6.0% premix.

[0013] Preferably, the mass ratio of the premix includes: vitamin A 6000IU, vitamin D3 2100IU, vitamin E20IU, vitamin B1 2mg, vitamin B2 4.5mg, vitamin K 1.5mg, copper 8mg, iron 100mg, manganese 60mg, zinc 80mg, iodine 0.8mg, and selenium 0.2mg.

[0014] Preferably, the mass percentage composition of the basic diet A is: 53.5% corn, 34.2% soybean meal, 5.0% wheat bran, 1.0% corn gluten meal, 1.3% soybean oil, and 5.0% premix.

[0015] Preferably, the mass percentage composition of the basic diet B is: 60-65% corn, 20-25% soybean meal, 4.0-6.0% wheat bran, 4.0-6.0% corn gluten meal, 1.0-2.0% soybean oil, and 4.0-6.0% premix.

[0016] Preferably, the mass ratio of the premix is: vitamin A 6000IU, vitamin D3 2100IU, vitamin E 20IU, vitamin B1 2mg, vitamin B2 4.5mg, vitamin K 1.5mg, copper 8mg, iron 100mg, manganese 60mg, zinc 80mg, iodine 0.8mg, and selenium 0.2mg.

[0017] Preferably, the mass percentage composition of the basic diet B is: 60.7% corn, 23.5% soybean meal, 5.0% wheat bran, 4.0% corn gluten meal, 1.8% soybean oil, and 5.0% premix.

[0018] Preferably, ornithine aspartate is used as a feed additive in poultry and aquatic animal feed.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] 1. The present invention adds ornithine aspartate as a feed additive to the daily diet to feed yellow-feathered broilers. The broilers are divided according to their age, and feeds with different properties and components are fed according to different ages. By adding ornithine aspartate to the feed, the growth performance of broilers can be improved, serum biochemical indicators can be improved, serum antioxidant capacity can be improved, and intestinal morphology and structure can be improved.

[0021] 2. In the present invention, the average daily feed intake and average daily weight gain of broilers supplemented with ornithine aspartate tend to increase, and the liver index of the broilers is significantly reduced, the CAT activity in the serum is significantly increased, the GSH-PX activity in the serum of the broilers is significantly increased, the glucose content and triglyceride content in the serum of the broilers are significantly reduced, the high-density lipoprotein content is significantly increased, the jejunal villus height is significantly increased, the ratio of villus height to crypt depth is significantly increased, the mRNA expression level of Occludin in the jejunum of broilers is significantly increased, the mRNA expression level of FAS is significantly decreased, the mRNA expression level of GLUT2 in the jejunum of broilers is significantly decreased, and the mRNA expression level of MTL is significantly increased. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a graph showing the effect of adding ornithine aspartate to the diet on the jejunum morphology of yellow-feathered broiler chickens according to an embodiment of the present invention, wherein a is the control group, b is the guanidine acetate group, and c is the ornithine aspartate group;

[0023] Figure 2 This is a diagram showing the effect of adding ornithine aspartate to the diet on the ileum morphology of yellow-feathered broiler chickens according to an embodiment of the present invention, wherein a is the control group, b is the guanidine acetate group, and c is the ornithine aspartate group. DETAILED DESCRIPTION

[0024] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0025] The inventors of this application have discovered that when raising broiler chickens, muscle energy supply is very important for fast-growing broilers. In order to ensure rapid growth of broilers, improving the intestinal morphology of broilers or maintaining intestinal homeostasis is a relatively good solution. Current research has found that there is no better way to improve the intestinal morphology of broilers.

[0026] In view of this, the present application provides the use of ornithine aspartate as a feed additive in yellow-feathered broiler diets. In this example, ornithine aspartate is added to the diet to explore the effects of ornithine aspartate on the growth performance, serum biochemical indicators, and intestinal morphology of yellow-feathered broiler chickens.

[0027] The present application uses ornithine aspartate as a feed additive and feeds broilers, which can improve feed absorption and serum biochemical indicators, enhance serum antioxidant capacity, and improve intestinal morphology and structure.

[0028] To verify the effects of the above examples, the experimental materials and reagents used in the following examples were commercially available unless otherwise specified. Where specific techniques or conditions are not specified in the examples, they can be performed using techniques or conditions described in literature in the art or according to the product instructions.

[0029] 1. Materials and Methods

[0030] 1.1 Experimental Animals and Experimental Design

[0031] The study used a completely randomized design. 180 one-day-old male yellow-feathered broiler chickens were randomly assigned to three treatment groups (control, GAA, and ORN-A), with six replicates per group and ten birds per replicate. The control group was fed a basal diet, while the GAA group received a basal diet supplemented with 500 mg / kg GAA, and the ORN-A group received a basal diet supplemented with 300 mg / kg ORN-A, the dosage of which had been determined in a preliminary study. The 42-day trial was divided into an early (1-21 days of age) and a late (22-42 days of age) trial.

[0032] 1.2 Experimental diet and feeding management

[0033] The henhouses and cages were flushed and disinfected before the experiment. During the experiment, broilers had free access to food and water. The experimental basal diet was formulated as a powdered feed based on the nutritional requirements of broilers as specified in the NRC (1994). The experimental diet formula and nutrient levels are shown in Table 1.

[0034] Table 1 Composition and nutritional levels of experimental diets

[0035]

[0036]

[0037] Note: The premix provides vitamin A 6000IU, vitamin D3 2100IU, vitamin E 20IU, vitamin B12mg, vitamin B 24.5mg, vitamin K 1.5mg, copper 8mg, iron 100mg, manganese 60mg, zinc 80mg, iodine 0.8mg, and selenium 0.2mg per kilogram of feed.

[0038] 1.3 Sample Collection

[0039] At 42 days of age, one broiler chicken near the mean weight in each replicate was selected for blood sampling and slaughtered. Serum was obtained from the blood sample after centrifugation and stored at −20°C for subsequent biochemical analysis. The heart, liver, spleen, and bursa of Fabricius were isolated and weighed on an electronic balance. Approximately 2 cm long segments were excised from the midsection of the jejunum and ileum, rinsed with saline, and stored in 4% paraformaldehyde for intestinal morphology. Approximately 1 cm long segments of the remaining jejunum were cut and stored at −80°C for gene expression analysis.

[0040] 1.4. Index determination

[0041] 1.4.1 Growth performance

[0042] The feed intake of yellow-feathered broilers was recorded daily. On days 1, 21, and 42 of the experiment, the yellow-feathered broilers of all experimental groups were weighed, and the average daily weight gain, average daily feed intake, and feed-to-weight ratio of each group were calculated.

[0043] Average daily feed intake (g) = total feed intake during the experimental period / (number of experimental chickens × number of experimental days);

[0044] Average daily weight gain (g) = (weight at the end of the test period - weight at the beginning) / (number of test chickens × number of test days);

[0045] Feed-to-weight ratio = average daily feed intake / average daily weight gain.

[0046] 1.4.2 Organ Index

[0047] The broiler weight was recorded before slaughter, and the heart, liver, spleen and bursa of Fabricius were removed after dissection to calculate the organ index according to the following formula;

[0048] Organ index (g / kg) = organ weight / broiler live weight.

[0049] 1.4.3 Serum biochemical indicators

[0050] Serum glutathione peroxidase (GSH-PX) activity, superoxide dismutase (T-SOD) activity, catalase (CAT) activity, and malondialdehyde (MDA) content, as well as serum glucose (GLU), triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) levels were detected using kits produced by Nanjing Jiancheng Bioengineering Research Institute.

[0051] 1.4.4 Observation of jejunum and ileum tissue morphology

[0052] The jejunum and ileum preserved in 4% paraformaldehyde solution were prepared into paraffin sections and stained with HE. The morphological indicators of the jejunum and ileum were observed under a microscope. The intact and straight villi were selected, and the villus height, crypt depth and the ratio of villus height to crypt depth (V / C) were counted and calculated.

[0053] 1.4.5. Jejunum-related gene expression determination

[0054] Jejunal RNA was extracted using FreeZol Reagent RNA extraction reagent. The extracted RNA was reverse transcribed into cDNA using a kit (HiScript III RT SuperMix for qPCR) and then subjected to RT-qPCR. All procedures were performed strictly according to the manufacturer's instructions. All kits used were purchased from Nanjing Novozymes Biotechnology Co., Ltd., and primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd. The genes assayed included claudin-1, occludens-1 (ZO-1), occludin, glucose transporter-2 (GLUT2), sodium / glucose cotransporter-1 (SGLT1), fatty acid synthase (FAS), and motilin (MTL). Primer information is provided in Table 2.

[0055] Table 2 Gene information and primer sequences

[0056]

[0057]

[0058] 1.5 Data processing and statistical analysis

[0059] The experimental data were organized in Excel 2016 and analyzed for variance using the one-way ANOVA procedure in SPSS 20.0 software. When the differences were significant, Duncan's method was used for multiple comparisons. All experimental data were expressed as "mean ± standard deviation", and P < 0.05 was used as the criterion for judging significant differences.

[0060] 2. Results and Analysis

[0061] 2.1 Effects of dietary ornithine aspartate supplementation on growth performance and organ indices in yellow-feathered broilers

[0062] 2.1.1 Effects of dietary ornithine aspartate supplementation on growth performance of yellow-feathered broilers

[0063] As shown in Table 3, at 1-21 days of age, the addition of guanidine acetate and ornithine aspartate to the diet had no significant effect on the average daily gain, average daily feed intake and feed-to-weight ratio of yellow-feathered broilers (P>0.05); at 22-42 days of age, compared with the control group, the addition of guanidine acetate to the diet could significantly increase the average daily gain and average daily feed intake of yellow-feathered broilers (P<0.05); at 1-42 days of age, compared with the control group, the addition of guanidine acetate to the diet could significantly increase the average daily gain of yellow-feathered broilers (P<0.05); the addition of guanidine acetate to the diet could significantly increase the body weight of yellow-feathered broilers at 42 days of age (P<0.05); the addition of guanidine acetate and ornithine aspartate to the diet had no significant effect on the feed-to-weight ratio of yellow-feathered broilers (P>0.05).

[0064] Table 3 Effects of dietary ornithine aspartate supplementation on growth performance of yellow-feathered broilers

[0065]

[0066]

[0067] Note: Data in the same row do not contain the same lowercase letters, which indicates significant differences (P<0.05); data with the same lowercase letters or no letters indicate no significant differences (P>0.05); the same applies to the following tables.

[0068] 2.1.2 Effects of dietary ornithine aspartate supplementation on organ indices in yellow-feathered broilers

[0069] As shown in Table 4, compared with the control group, the addition of guanidine acetate and ornithine aspartate to the diet had no significant effect on the heart index, spleen index and bursa index of yellow-feathered broilers (P>0.05), but significantly reduced the liver index of yellow-feathered broilers (P<0.05).

[0070] Table 4 Effects of dietary ornithine aspartate supplementation on organ indices of yellow-feathered broilers

[0071]

[0072]

[0073] 2.2 Effects of dietary ornithine aspartate supplementation on serum biochemical parameters in yellow-feathered broilers

[0074] 2.2.1 Effect of dietary ornithine aspartate supplementation on serum antioxidant capacity in yellow-feathered broilers

[0075] As shown in Table 5, compared with the control group, the addition of guanidine acetate and ornithine aspartate to the diet had no significant effect on the level of T-SOD and the content of MDA in the serum of yellow-feathered broilers (P>0.05); the addition of guanidine acetate and ornithine aspartate to the diet could significantly increase the CAT activity in the serum of yellow-feathered broilers (P<0.05); the addition of ornithine aspartate to the diet could significantly increase the GSH-PX activity in the serum of yellow-feathered broilers (P<0.05).

[0076] Table 5 Effects of dietary ornithine aspartate supplementation on serum antioxidant capacity of yellow-feathered broilers

[0077]

[0078] 2.2.2 Effects of dietary ornithine aspartate supplementation on serum biochemical parameters in yellow-feathered broilers

[0079] As shown in Table 6, compared with the control group, the addition of guanidine acetate and ornithine aspartate to the diet had no significant effect on the total cholesterol content and low-density lipoprotein content in the serum of yellow-feathered broilers (P>0.05); the addition of guanidine acetate and ornithine aspartate to the diet could significantly reduce the glucose content and triglyceride content in the serum of yellow-feathered broilers (P<0.05), and significantly increase the high-density lipoprotein content in the serum of yellow-feathered broilers (P<0.05).

[0080] Table 6 Effects of adding ornithine aspartate to the diet on serum biochemical parameters of yellow-feathered broilers

[0081]

[0082]

[0083] 2.3 Effects of dietary ornithine aspartate supplementation on intestinal tissue morphology in yellow-feathered broilers

[0084] As shown in Table 7, compared with the control group, the addition of guanidine acetate and ornithine aspartate to the diet had no significant effect on the morphology of the ileum (P>0.05); the addition of guanidine acetate and ornithine aspartate to the diet significantly increased the height of the villi in the jejunum of yellow-feathered broilers (P<0.05) and significantly increased the ratio of the villi height to the crypt depth in the jejunum of yellow-feathered broilers (P<0.05). The tissue sections of the jejunum and ileum of the three groups of yellow-feathered broilers were shown in Table 7. Figure 1 、 Figure 2 .

[0085] Table 7 Effects of dietary ornithine aspartate supplementation on intestinal tissue morphology in yellow-feathered broilers

[0086]

[0087] 2.4 Effects of dietary ornithine aspartate supplementation on jejunal gene expression in yellow-feathered broiler chickens

[0088] As shown in Table 8, compared with the control group, the addition of guanidine acetate and ornithine aspartate to the diet had no significant effect on the mRNA expression of Claudin-1 and ZO-1 in the jejunum of yellow-feathered broilers (P>0.05), but significantly increased the mRNA expression of Occludin (P<0.05); the addition of ornithine aspartate to the diet significantly reduced the mRNA expression of glucose transporter GLUT2 in the jejunum of yellow-feathered broilers (P<0.05), and the addition of guanidine acetate and ornithine aspartate had no significant effect on the mRNA expression of glucose transporter SGLT1 in the jejunum (P>0.05); the addition of guanidine acetate and ornithine aspartate significantly reduced the mRNA expression of fatty acid synthase FAS in the jejunum (P<0.05); the addition of ornithine aspartate significantly increased the mRNA expression of motilin MTL in the jejunum (P<0.05).

[0089] Table 8 Effects of dietary ornithine aspartate supplementation on gene expression levels in the jejunum of yellow-feathered broiler chickens

[0090]

[0091]

[0092] In summary, the present invention can improve the growth performance of broiler chickens, improve serum biochemical indicators, improve serum antioxidant capacity, and improve intestinal morphology and structure by adding 300-1500 mg / kg ornithine aspartate to the daily diet of yellow-feathered broiler chickens.

[0093] The embodiments disclosed in the present invention are preferred embodiments, but are not limited to them. Ordinary technicians in this field can easily understand the spirit of the present invention based on the above embodiments and make different extensions and changes. As long as they do not deviate from the spirit of the present invention, they are all within the scope of protection of the present invention.

Claims

1. Use of ornithine aspartate in preparing a broiler feed additive, characterized in that: The feed additive is used to improve feed absorption and serum biochemical indicators, increase serum antioxidant capacity, and improve intestinal morphology and structure; The feed additive is used in a single dosage of 300-1500 mg / kg feed.

2. The use according to claim 1, characterized in that Broilers were divided based on their age. Different feeds with different properties and components were fed according to their age. During the experiment, broilers had free access to food and water. Wherein, the feeds with different properties and components fed to pigs at different ages are: (1) Male yellow-feathered broiler chickens, aged 1 to 21 days, were fed a crushed pelleted diet comprising a basal diet A and a feed additive; (2) At 22-42 days of age, male yellow-feathered broiler chickens are fed a diet comprising a basic diet B and feed additives.

3. The use according to claim 2, characterized in that The mass percentage composition of the basic diet A is: 50-55% corn, 30-35% soybean meal, 4.0-6.0% wheat bran, 0.5-2.0% corn gluten meal, 0.5-2.0% soybean oil, and 4.0-6.0% premix.

4. The use according to claim 3, characterized in that The premix comprises the following raw materials: 6000 IU of vitamin A, 2100 IU of vitamin D3, 20 IU of vitamin E, 2 mg of vitamin B1, 4.5 mg of vitamin B2, 1.5 mg of vitamin K, 8 mg of copper, 100 mg of iron, 60 mg of manganese, 80 mg of zinc, 0.8 mg of iodine, and 0.2 mg of selenium.

5. The use according to claim 3, characterized in that The mass percentages of the basic diet A include: 53.5% corn, 34.2% soybean meal, 5.0% wheat bran, 1.0% corn gluten meal, 1.3% soybean oil, and 5.0% premix.

6. The use according to claim 2, characterized in that: The mass percentage composition of the basic diet B is: 60-65% corn, 20-25% soybean meal, 4.0-6.0% wheat bran, 4.0-6.0% corn gluten meal, 1.0-2.0% soybean oil, and 4.0-6.0% premix.

7. The use according to claim 6, characterized in that: The premix comprises the following raw materials: 6000 IU of vitamin A, 2100 IU of vitamin D3, 20 IU of vitamin E, 2 mg of vitamin B1, 4.5 mg of vitamin B2, 1.5 mg of vitamin K, 8 mg of copper, 100 mg of iron, 60 mg of manganese, 80 mg of zinc, 0.8 mg of iodine, and 0.2 mg of selenium.

8. The use according to claim 6, characterized in that: The weight percentage composition of the basic diet B is: 60.7% corn, 23.5% soybean meal, 5.0% wheat bran, 4.0% corn gluten meal, 1.8% soybean oil, and 5.0% premix.

9. The use according to any one of claims 1 to 8, characterized in that: The broiler chicken is a yellow-feathered broiler chicken.

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