Feed for improving estrus rate of ewes and preparation method thereof

Abstract

The invention provides a feed for improving estrus rate of ewes and a preparation method thereof, and belongs to the technical field of feeds. The preparation method comprises the following steps: fermenting herba epimedii, radix morindae officinalis, herba leonuri and Chinese chive seeds with selenium-enriched saccharomycetes and manganese-enriched saccharomycetes, carrying out alcohol precipitation to obtain a solid and a liquid, adding a solid extraction agent into the liquid for extraction to obtain an extract, adding the extract and the compound polysaccharide into a basic ration, and uniformly stirring and mixing to obtain the feed for improving the estrus rate of the ewes. The feed for improving estrus rate of ewes is added with traditional Chinese medicine components, is safe, free of side effects, natural and not easy to generate drug resistance, and has a relatively good estrus promoting effect; the feed additive has multiple effects of enhancing the anti-stress ability of livestock and poultry, improving the immunity of the organism, improving the quality of livestock and poultry products, regulating the metabolism of the organism, promoting the growth and development of animals, inhibiting the growth of bacteria, enhancing the antioxidant ability and the like, and has a wide application prospect.

Description

Technical Field

[0001] The invention relates to the technical field of feed, and in particular to a feed for improving the estrus rate of ewes and a preparation method thereof. Background Art

[0002] With the rapid development of the feed additive industry, the hazards brought by chemical additives have become a consensus, such as drug residues, drug resistance, carcinogenicity, teratogenicity, mutagenicity and other side effects, which have forced people to consider carefully when choosing additives. In order to explore the source of feed additives, many scholars have explored multiple channels. Nowadays, Chinese herbal feed additives are favored by people for their unique effects, good effects, no residues, no drug resistance and no pollution. Chinese herbal feed additives refer to pure natural feed additives made by taking the traditional Chinese theory of the physical properties, taste and relationship between Chinese herbal medicines as the main guide, supplemented by the theoretical technology of breeding and feed industry. In the development of feed additives, it is favored for its unique advantages and has become one of the hot spots of research and development.

[0003] At present, the means to improve the reproductive performance of ewes are mostly synchronized estrus technology, because synchronized estrus technology is convenient for centralized artificial insemination; saving breeding sheep, labor and time; improving the utilization of excellent germplasm resources; facilitating the batch production of commercial sheep; and because the lambing time is uniform and the specifications are relatively consistent, it is convenient for the factory production of the mutton industry. The methods of synchronized estrus technology include extending the luteal phase and shortening the luteal phase. Extending the luteal phase mainly uses drugs that inhibit the growth and development of follicles, which can inhibit the growth, development, maturation and ovulation of follicles on the ovaries of female animals, but does not affect the natural regression of the corpus luteum, such as progesterone, medroxyprogesterone, and four-deoxyglucose. Shortening the luteal phase is to inject a hormone that can dissolve the corpus luteum when the ovaries of female animals are in the luteal phase. This hormone can dissolve the corpus luteum at different periods at the same time, so as to achieve the purpose of synchronized estrus of female animals, such as prostaglandin F2α (PGF2α). In addition, gonadotropins, such as pregnant mare blood hormone, chorionic hormone, follicle-stimulating hormone, luteinizing hormone and gonadotropin-releasing hormone, can also be used to promote ovulation and increase conception rate. In production practice, due to the high price of hormones required for estrus synchronization technology and the high labor costs required for operations such as plugging, the labor costs required in large-scale farms in the future will be even higher.

[0004] Therefore, developing a pure natural feed that can increase the estrus rate of ewes will have broad application prospects. Summary of the invention

[0005] The purpose of the present invention is to provide a feed for improving the estrus rate of ewes and a preparation method thereof. By adding traditional Chinese medicine components, the feed is safe and has no side effects, is naturally not easy to produce drug resistance, has a good aphrodisiac effect, can enhance the anti-stress ability of livestock and poultry, improve the body's immunity, improve the quality of livestock and poultry products, regulate the body's metabolism, promote animal growth and development, inhibit bacterial growth, enhance antioxidant capacity and other multiple functions, and has broad application prospects.

[0006] The technical solution of the present invention is achieved in this way: The invention provides a method for preparing a feed capable of improving the estrus rate of ewes. The method comprises the following steps: fermenting epimedium, Morinda officinalis, Leonurus japonicus and leek seeds with selenium-enriched yeast and manganese-enriched yeast, and then precipitating the solid and liquid through alcohol, wherein the solid is a composite polysaccharide, and a solid extractant is added into the liquid to obtain an extract, which is added into a basic diet together with the composite polysaccharide, and stirred and mixed evenly to obtain the feed capable of improving the estrus rate of ewes.

[0007] As a further improvement of the present invention, the following steps are included: S1. The selenium-enriched yeast and the manganese-enriched yeast were inoculated into a malt agar medium, activated and cultured to obtain a bacterial seed solution; S2. Wash, dry, and crush Epimedium, Morinda officinalis, Leonurus japonicus, and leek seeds, add them to water, add selenium-rich yeast and manganese-rich yeast seed solution, ferment and culture, filter, and retain the filtrate; S3. Add ethanol to the filtrate for precipitation, centrifuge, wash the solid, dry, and obtain a complex polysaccharide, and retain the centrifuge; S4. The ethanol is recovered from the centrifuged liquid, a solid extractant is added to the remaining liquid, adsorbed by stirring under an external magnetic field, separated by a magnet, the solid is added to ethanol, heated and stirred for desorption, the solid is separated by a magnet, recycled, the solvent is removed from the liquid under reduced pressure, and dried to obtain an extract; S5. Add soybean isoflavones, complex polysaccharides and extracts to the basic diet, stir and mix evenly, and prepare feed for improving the estrus rate of ewes.

[0008] As a further improvement of the present invention, the activation culture conditions in step S1 are 28-32°C, 150-250r / min, activation culture 18-24h, and the bacterial content of the bacterial seed liquid is 10 8 -10 9 cfu / mL.

[0009] As a further improvement of the present invention, the mass ratio of epimedium, Morinda officinalis, Leonurus japonicus and leek seeds in step S2 is 10:4-6:2-3:3-5, the inoculation amounts of the selenium-enriched yeast and manganese-enriched yeast seed solutions are 1-2 v / v% and 0.5-1.5 v / v%, respectively, and the fermentation and culture conditions are 28-32°C, 150-250 r / min, and the fermentation and culture is 48-72 h.

[0010] As a further improvement of the present invention, in step S3, ethanol is added until the ethanol content of the system is 80-85wt%, and the precipitation time is 1-2h.

[0011] As a further improvement of the present invention, the preparation method of the solid extractant in step S4 is as follows: T1. Add silica microspheres to water, add ferric chloride and ferrous chloride, add ammonia water under inert gas protection, heat and stir to react, centrifuge, wash, dry, and calcine to obtain magnetic silica microspheres; T2. adding magnetic silica microspheres, phytic acid, β-cyclodextrin, and potassium dihydrogen phosphate into water, performing hydrothermal reaction, separating with a magnet, washing, and drying to obtain a magnetic silica-phytic acid-cyclodextrin composite material; T3. Add the magnetic silica-phytic acid-cyclodextrin composite material into Tris-HCl solution, add dopamine hydrochloride, heat and stir to react, separate with a magnet, wash, and dry to obtain a solid extractant.

[0012] As a further improvement of the present invention, the particle size of the silica microspheres in step T1 is 200-500nm, the mass ratio of the silica microspheres, ferric chloride, ferrous chloride and ammonia water is 10-12:3.24:1.26:4-6, the temperature of the heating and stirring reaction is 75-85°C, the time is 3-5h, the calcination temperature is 400-500°C, and the time is 0.5-1.5h; the magnetic silica microspheres, The mass ratio of phytic acid, β-cyclodextrin and potassium dihydrogen phosphate is 10:1-2:4-6:1-2, the temperature of the hydrothermal reaction is 140-160°C, and the time is 9-11h; the pH value of the Tris-HCl solution in step T3 is 8.5-9.5, the mass ratio of the magnetic silica-phytic acid-cyclodextrin composite material and dopamine hydrochloride is 10:1-2, the temperature of the heating and stirring reaction is 40-50°C, and the time is 1-3h.

[0013] As a further improvement of the present invention, the liquid-to-solid ratio of the liquid and solid extractants in step S4 is 5-10 mL:1 g, the strength of the external magnetic field is 0.05-0.1 T, the stirring adsorption time is 30-50 min, the heating temperature is 40-50 ° C, and the stirring desorption time is 10-20 min.

[0014] As a further improvement of the present invention, the basic diet in step S5 is mixed by the following raw materials in parts by weight: 10-20 parts of wheat straw, 4-6 parts of alfalfa, 20-25 parts of refined grain supplement, 20-30 parts of straw, and 15-20 parts of sweet potato starch; the refined grain supplement is mixed by the following raw materials in parts by weight: 10-30 parts of distiller's grains, 5-15 parts of soybean meal, 3-7 parts of vegetable oil, 1-3 parts of molasses, 1-2 parts of salt, 0.5-1.5 parts of calcium phosphate, 1-2 parts of vitamin C, 0.5-1.5 parts of vitamin A, and 0.3-0.5 parts of vitamin E; the mass ratio of the basic diet, soybean isoflavones, complex polysaccharides and extracts is 100:0.5-1:8-10:10-12.

[0015] The present invention further protects a feed for improving the estrus rate of ewes prepared by the above preparation method.

[0016] The present invention has the following beneficial effects: Epimedium, Morinda officinalis and leek seeds are rich in vitamin E and selenium, which can improve the reproductive function of animals, and Leonurus japonicus contains vitamin A-like substances and leonurine, vitamin A can maintain the integrity of reproductive epithelium, while leonurine has a strong contraction effect on the uterus and promotes ovarian blood circulation, and also has an anti-inflammatory effect on the uterus. Therefore, the Chinese medicine component of the present invention has a good effect on the reproductive system and aphrodisiac.

[0017] The present invention uses selenium-enriched yeast and manganese-enriched yeast to ferment the Chinese medicine components. The obtained fermentation product is rich in organic selenium and organic manganese, which improves the absorption rate of animals to selenium and manganese. Selenium has a significant effect on the reproductive capacity of animals. It can affect the development of animal reproductive organs, regulate the estrus and ovulation time of animals, improve the semen quality and sperm vitality of male animals, ensure the conception and embryonic development of female animals, etc., thereby improving the reproductive capacity of animals. For female animals, selenium deficiency will cause the mother animals to show irregular estrus or estrus, thereby reducing their conception rate. Manganese exists mainly in the form of divalent or trivalent compounds in animals, and can improve the immune efficacy of the body by increasing the activity of multiple enzymes in nonspecific immunity, thereby indirectly affecting the reproductive capacity of animals. Manganese can regulate the level of interferon (IFN) in the body, and interferon can enhance the phagocytic ability of macrophages, thereby regulating protein synthesis. The two have a synergistic effect.

[0018] The Chinese medicine component of the present invention has naturalness as feed additive, and avoids the generation of drug resistance. It also can increase the nutrition of livestock products at the same time, play a certain health care effect, contain rich alkaloids, flavonoids, quinones, saponins, organic acids and volatile components and other organic substances, can enhance the anti-stress ability of livestock and poultry, improve body immunity, improve livestock and poultry product quality, regulate body metabolism, promote animal growth and development, inhibit bacterial growth, enhance antioxidant capacity and other multiple effects, the Chinese medicine component of the present invention has a promoting effect on the reproductive hormone secretion, estrus ovulation, pregnancy and childbirth and other breeding-related processes of livestock and poultry, wherein, organic components such as flavonoids can participate in the reproductive hormone secretion system, and then play an active role in the maturation of gonadal development and reproductive function of animals. However, if the materials such as this type of alkaloids, flavonoids, quinones, saponins, organic acids and volatile components are obtained with high purity, it becomes a problem.

[0019] The present invention prepares a magnetic solid extractant, which has the characteristics of good dispersibility, biocompatibility, large specific surface area, superparamagnetism and magnetic responsiveness, etc., and selectively combines with the specific sites of the above-mentioned components of traditional Chinese medicine through physical, chemical adsorption (such as electrostatic interaction, hydrogen bond, covalent bond and coordination) or bioaffinity. Under the action of an external magnetic field, the target components can be quickly concentrated or separated, thereby achieving separation and purification. Due to the presence of superparamagnetism and magnetic responsiveness, the particles can be quickly separated under an external magnetic field, thereby improving the efficiency of extraction and separation. However, pure magnetic particles are prone to coagulation. The present invention introduces amino, carboxyl, hydroxyl, silicon dioxide and the like through polydopamine modification to modify the surface of the magnetic solid extractant, effectively avoiding the aggregation and oxidation of the magnetic particles, and promoting its combination with the components of traditional Chinese medicine. The composite material obtained by the reaction of hydroxyl groups on silica, phytic acid and β-cyclodextrin has good adsorption effect. The hydrophobic cavity of β-cyclodextrin significantly improves the adsorption effect of fat-soluble active substances, and phytic acid can form hydrogen bonds through its phosphate group to fix active molecules, thereby greatly improving the adsorption and separation efficiency of active components.

[0020] Soy isoflavones can not only promote animal growth and mammary gland development, but also enhance the animal's immune function and improve the disease resistance of livestock and poultry. They have obvious aphrodisiac effects and can improve the overall reproductive performance of animals by increasing the number of ovulations, litter size and lactation capacity.

[0021] The feed for improving the estrus rate of ewes of the present invention is safe and has no side effects by adding traditional Chinese medicine components, is naturally not easy to produce drug resistance, has a good aphrodisiac effect, can enhance the anti-stress ability of livestock and poultry, improve the immunity of the body, improve the quality of livestock and poultry products, regulate the metabolism of the body, promote animal growth and development, inhibit bacterial growth, enhance antioxidant capacity and other multiple functions, and has broad application prospects. DETAILED DESCRIPTION

[0022] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] Selenium-enriched yeast, 20 billion cfu / g, was purchased from Angel Yeast Co., Ltd. Manganese-enriched yeast, 10 billion cfu / g, was purchased from Jiangsu Yihaotian Biotechnology Co., Ltd.

[0024] Preparation Example 1 Preparation of solid extractant Here’s how: T1. Add 10g silica microspheres to 200mL water, add 3.24g ferric chloride and 1.26g ferrous chloride, add 4g ammonia water under nitrogen protection, heat to 75℃, stir and react for 3h, centrifuge, wash, dry, and calcine at 400℃ for 0.5h to obtain magnetic silica microspheres; T2. Add 10g magnetic silica microspheres, 1g phytic acid, 4g β-cyclodextrin, and 1g potassium dihydrogen phosphate into 200mL water, heat to 140°C, perform hydrothermal reaction for 9h, separate with a magnet, wash, and dry to obtain a magnetic silica-phytic acid-cyclodextrin composite material; T3. Add 10g of magnetic silica-phytic acid-cyclodextrin composite material to 100mL of Tris-HCl solution with a pH value of 8.5, add 1g of dopamine hydrochloride, heat to 40°C, stir to react for 1h, separate with a magnet, wash, and dry to obtain a solid extractant.

[0025] Preparation Example 2 Preparation of solid extractant Here’s how: T1. Add 12g silica microspheres to 200mL water, add 3.24g ferric chloride and 1.26g ferrous chloride, add 6g ammonia water under nitrogen protection, heat to 85℃, stir and react for 5h, centrifuge, wash, dry, and calcine at 500℃ for 1.5h to obtain magnetic silica microspheres; T2. 10 g of magnetic silica microspheres, 2 g of phytic acid, 6 g of β-cyclodextrin, and 2 g of potassium dihydrogen phosphate were added to 200 mL of water, heated to 160°C, hydrothermally reacted for 11 h, separated by a magnet, washed, and dried to obtain a magnetic silica-phytic acid-cyclodextrin composite material; T3. Add 10g of magnetic silica-phytic acid-cyclodextrin composite material to 100mL of Tris-HCl solution with a pH value of 9.5, add 2g of dopamine hydrochloride, heat to 50°C, stir and react for 3h, separate with a magnet, wash, and dry to obtain a solid extractant.

[0026] Preparation Example 3 Preparation of solid extractant Here’s how: T1. Add 11g silica microspheres to 200mL water, add 3.24g ferric chloride and 1.26g ferrous chloride, add 5g ammonia water under nitrogen protection, heat to 80℃, stir and react for 4h, centrifuge, wash, dry, and calcine at 450℃ for 1h to obtain magnetic silica microspheres; T2. 10 g of magnetic silica microspheres, 1.5 g of phytic acid, 5 g of β-cyclodextrin, and 1.2 g of potassium dihydrogen phosphate were added to 200 mL of water, heated to 150°C, hydrothermally reacted for 10 h, separated by a magnet, washed, and dried to obtain a magnetic silica-phytic acid-cyclodextrin composite material; T3. Add 10 g of magnetic silica-phytic acid-cyclodextrin composite material to 100 mL of Tris-HCl solution with a pH value of 9, add 1.5 g of dopamine hydrochloride, heat to 45°C, stir to react for 2 h, separate with a magnet, wash, and dry to obtain a solid extractant.

[0027] Comparative Preparation Example 1 Compared with Preparation Example 3, the difference is that step T2 is not performed.

[0028] The details are as follows: T1. Add 11g silica microspheres to 200mL water, add 3.24g ferric chloride and 1.26g ferrous chloride, add 5g ammonia water under nitrogen protection, heat to 80℃, stir and react for 4h, centrifuge, wash, dry, and calcine at 450℃ for 1h to obtain magnetic silica microspheres; T2. Add 10 g of magnetic silica microspheres into 100 mL of Tris-HCl solution with a pH value of 9, add 1.5 g of dopamine hydrochloride, heat to 45°C, stir to react for 2 h, separate with a magnet, wash, and dry to obtain a solid extractant.

[0029] Comparative Preparation Example 2 Compared with Preparation Example 3, the difference is that step T3 is not performed.

[0030] The details are as follows: T1. Add 11g silica microspheres to 200mL water, add 3.24g ferric chloride and 1.26g ferrous chloride, add 5g ammonia water under nitrogen protection, heat to 80℃, stir and react for 4h, centrifuge, wash, dry, and calcine at 450℃ for 1h to obtain magnetic silica microspheres; T2. Add 10g magnetic silica microspheres, 1.5g phytic acid, 5g β-cyclodextrin, and 1.2g potassium dihydrogen phosphate into 200mL water, heat to 150°C, perform hydrothermal reaction for 10h, separate with a magnet, wash, and dry to obtain a magnetic silica-phytic acid-cyclodextrin composite material, which is a solid extractant.

[0031] Embodiment 1: This embodiment provides a method for preparing a feed for improving the estrus rate of ewes, comprising the following steps: S1. Selenium-enriched yeast and manganese-enriched yeast were inoculated into malt extract agar medium and cultured at 32°C, 150 r / min for 18 h to obtain a culture medium with a bacterial count of 10 8 -10 9 cfu / mL bacterial seed solution; S2. Wash, dry and crush 10g of Epimedium, 4g of Morinda officinalis, 2g of Leonurus japonicus and 3g of leek seeds respectively, add them into 200mL of water, add selenium-enriched yeast and manganese-enriched yeast seed solution, the inoculation amount is 1v / v% and 0.5v / v%, respectively, at 28℃, 150-250r / min, ferment and culture for 48h, filter and retain the filtrate; S3. Ethanol was added to the filtrate until the ethanol content of the system was 80wt%, precipitated for 1h, centrifuged, the solid was washed and dried to obtain a composite polysaccharide, and the centrifuge was retained; S4. Recover ethanol from the centrifuged liquid, add 10 g of the solid extractant prepared in Preparation Example 1 to 50 mL of the remaining liquid, stir and adsorb for 30 min under an external magnetic field with an intensity of 0.05 T, separate with a magnet, add the solid to ethanol, heat to 40° C., stir and desorb for 10 min, separate the solid with a magnet, recycle it, remove the solvent from the liquid under reduced pressure, and dry it to obtain an extract; S5. Add 0.5 g of soy isoflavones, 8 g of complex polysaccharides and 10 g of extract to 100 g of the basic diet, stir and mix for 30 min to prepare a feed for improving the estrus rate of ewes; The basic diet is prepared by mixing the following raw materials by weight: 15 parts of wheat straw, 5 parts of alfalfa, 22 parts of refined grain supplement, 25 parts of straw, and 17 parts of sweet potato starch; The refined grain supplement is prepared by mixing the following raw materials in parts by weight: 20 parts of distiller's grains, 10 parts of soybean meal, 5 parts of vegetable oil, 2 parts of molasses, 1.5 parts of salt, 1 part of calcium phosphate, 1.5 parts of vitamin C, 1 part of vitamin A, and 0.4 part of vitamin E.

[0032] Embodiment 2: This embodiment provides a method for preparing a feed for improving the estrus rate of ewes, comprising the following steps: S1. Selenium-enriched yeast and manganese-enriched yeast were inoculated into malt extract agar medium, and activated and cultured at 28°C, 250r / min for 24h to obtain a culture medium with a bacterial count of 10 8 -10 9 cfu / mL bacterial seed solution; S2. Wash, dry and crush 10g of Epimedium, 6g of Morinda officinalis, 3g of Leonurus japonicus and 5g of leek seeds respectively, add them into 200mL of water, add the seed solution of selenium-enriched yeast and manganese-enriched yeast, the inoculation amount is 2v / v% and 1.5v / v%, respectively, at 32℃, 250r / min, ferment and culture for 72h, filter and keep the filtrate; S3. Ethanol was added to the filtrate until the ethanol content of the system was 85wt%, precipitated for 2h, centrifuged, the solid was washed and dried to obtain a composite polysaccharide, and the centrifuge was retained; S4. Recover ethanol from the centrifuged liquid, add 10 g of the solid extractant prepared in Preparation Example 2 to 100 mL of the remaining liquid, stir and adsorb for 50 min under an external magnetic field with an intensity of 0.1 T, separate with a magnet, add the solid to ethanol, heat to 50° C., stir and desorb for 20 min, separate the solid with a magnet, recycle it, remove the solvent from the liquid under reduced pressure, and dry it to obtain an extract; S5. Add 1 g of soy isoflavones, 10 g of complex polysaccharides and 12 g of extract to 100 g of the basic diet, stir and mix for 30 min to prepare a feed for improving the estrus rate of ewes; The basic diet is prepared by mixing the following raw materials by weight: 15 parts of wheat straw, 5 parts of alfalfa, 22 parts of refined grain supplement, 25 parts of straw, and 17 parts of sweet potato starch; The refined grain supplement is prepared by mixing the following raw materials in parts by weight: 20 parts of distiller's grains, 10 parts of soybean meal, 5 parts of vegetable oil, 2 parts of molasses, 1.5 parts of salt, 1 part of calcium phosphate, 1.5 parts of vitamin C, 1 part of vitamin A, and 0.4 part of vitamin E.

[0033] Embodiment 3: This embodiment provides a method for preparing a feed for improving the estrus rate of ewes, comprising the following steps: S1. Selenium-enriched yeast and manganese-enriched yeast were inoculated into malt extract agar medium, and activated and cultured at 30°C, 200 r / min for 21 h to obtain a culture medium with a bacterial count of 10 8 -10 9 cfu / mL bacterial seed solution; S2. Wash, dry and crush 10g of Epimedium, 5g of Morinda officinalis, 2.5g of Leonurus japonicus and 4g of leek seeds respectively, add them into 200mL of water, add selenium-enriched yeast and manganese-enriched yeast seed solution, the inoculation amount is 1.5v / v% and 1v / v%, respectively, at 30℃, 200r / min, ferment and culture for 56h, filter and retain the filtrate; S3. Ethanol was added to the filtrate until the ethanol content of the system was 82wt%, precipitated for 1.5h, centrifuged, the solid was washed and dried to obtain a composite polysaccharide, and the centrifuge was retained; S4. Recover ethanol from the centrifuged liquid, add 10 g of the solid extractant prepared in Preparation Example 3 to 70 mL of the remaining liquid, stir and adsorb for 40 min under an external magnetic field with an intensity of 0.07 T, separate with a magnet, add the solid to ethanol, heat to 45° C., stir and desorb for 15 min, separate the solid with a magnet, recycle it, remove the solvent from the liquid under reduced pressure, and dry it to obtain an extract; S5. Add 0.7 g of soy isoflavones, 9 g of complex polysaccharides and 11 g of extract to 100 g of the basic diet, stir and mix for 30 min to prepare a feed for improving the estrus rate of ewes; The basic diet is prepared by mixing the following raw materials by weight: 15 parts of wheat straw, 5 parts of alfalfa, 22 parts of refined grain supplement, 25 parts of straw, and 17 parts of sweet potato starch; The refined grain supplement is prepared by mixing the following raw materials in parts by weight: 20 parts of distiller's grains, 10 parts of soybean meal, 5 parts of vegetable oil, 2 parts of molasses, 1.5 parts of salt, 1 part of calcium phosphate, 1.5 parts of vitamin C, 1 part of vitamin A, and 0.4 part of vitamin E.

[0034] Comparative Example 1 Compared with Example 3, the difference is that no selenium-enriched yeast seed liquid is inoculated in step S2.

[0035] The details are as follows: S2. Wash, dry and crush 10g of Epimedium, 5g of Morinda officinalis, 2.5g of Leonurus japonicus and 4g of leek seeds respectively, add them into 200mL of water, add manganese-rich yeast seed solution with an inoculation amount of 2.5v / v%, 30℃, 200r / min, ferment and culture for 56h, filter and retain the filtrate.

[0036] Comparative Example 2 Compared with Example 3, the difference is that the manganese-rich yeast seed liquid is not inoculated in step S2.

[0037] The details are as follows: S2. Wash, dry and crush 10g of Epimedium, 5g of Morinda officinalis, 2.5g of Leonurus japonicus and 4g of leek seeds respectively, add them into 200mL of water, add selenium-enriched yeast seed solution with an inoculation amount of 2.5v / v%, 30℃, 200r / min, ferment and culture for 56h, filter and retain the filtrate.

[0038] Comparative Example 3 Compared with Example 3, the difference is that in step S2, no selenium-enriched yeast and manganese-enriched yeast seed liquid is inoculated, and heating and boiling extraction is adopted.

[0039] The details are as follows: S2. Wash, dry and crush 10g of Epimedium, 5g of Morinda officinalis, 2.5g of Leonurus japonicus and 4g of leek seeds respectively, add them into 200mL of water, heat and boil to extract for 5h, filter and keep the filtrate.

[0040] Comparative Example 4 Compared with Example 3, the difference is that the solid extractant in step S4 is prepared by Comparative Preparation Example 1.

[0041] Comparative Example 5 Compared with Example 3, the difference is that the solid extractant in step S4 is prepared by Comparative Preparation Example 2.

[0042] Comparative Example 6 Compared with Example 3, the difference is that no external magnetic field is applied in step S4.

[0043] The details are as follows: S4. Recover ethanol from the centrifuged liquid, add 10 g of the solid extractant prepared in Preparation Example 3 to 70 mL of the remaining liquid, stir and adsorb for 40 min, separate with a magnet, add the solid to ethanol, heat to 45°C, stir and desorb for 15 min, separate the solid with a magnet and recycle it, remove the solvent from the liquid under reduced pressure, and dry to obtain an extract.

[0044] Comparative Example 7 Compared with Example 3, the difference is that no solid extractant is added in step S4, and extraction is performed with dichloromethane.

[0045] The details are as follows: S4. The ethanol is recovered from the centrifuged liquid, and 100 mL of dichloromethane is added to 70 mL of the remaining liquid for extraction. The liquids are separated, and the organic phase is decompressed to remove the solvent and dried to obtain an extract.

[0046] Test Example 1 The feeds for improving estrus rate of ewes prepared in Examples 1-3 and Comparative Examples 1-7 were tested for selenium and manganese content, and the results are shown in Table 1. Reference standards: "GB 5009.242-2017 National Food Safety Standard Determination of Manganese in Foods" and "GB5009.93-2017 National Food Safety Standard Determination of Selenium in Foods"; Table 1

[0047] It can be seen from the above table that the feeds for improving the estrus rate of ewes prepared in Examples 1-3 of the present invention have high contents of manganese and selenium.

[0048] Test Example 2 Healthy, obstetric disease-free small-tailed Han sheep empty-stomach ewes were randomly divided into a blank group, Example 1-3 groups, and Comparative Example 1-7 groups, with 20 ewes in each group, and the feed for improving the estrus rate of ewes prepared by Example 1-3 or Comparative Example 1-7 of the present invention was continuously fed for 15 days, and the blank group was fed with a basic diet (the basic diet was mixed with the following raw materials by weight: 15 parts of wheat straw, 5 parts of alfalfa, 22 parts of refined grain supplement, 25 parts of straw, and 17 parts of sweet potato starch; the refined grain supplement was mixed with the following raw materials by weight: 20 parts of distiller's grains, 10 parts of soybean meal, 5 parts of vegetable oil, 2 parts of molasses, 1.5 parts of salt, 1 part of calcium phosphate, 1.5 parts of vitamin C, 1 part of vitamin A, and 0.4 parts of vitamin E). Feed once at 8:00 and 15:00 every day, and drink water freely. During the experiment, the appetite, spirit, and diarrhea of ​​the test sheep were carefully observed every day, and records were kept; the sheep houses were disinfected regularly, and immunization and deworming were carried out according to routine procedures.

[0049] Record the amount of feed and the amount of leftover feed in the previous 15 days every day to calculate the feed intake. The amount of leftover feed per day should be 5%-10% of the feed amount. The feed amount can be adjusted appropriately according to the amount of leftover feed to accurately calculate the average daily dry matter intake of each group of sheep. The results are shown in Table 2.

[0050] Table 2

[0051] Notes: Compared with the blank group, P<0.05.

[0052] It can be seen from the above table that the feed for improving the estrus rate of ewes prepared in Examples 1-3 of the present invention significantly increased the food intake of ewes.

[0053] Estrus identification and breeding: Use rams to test estrus every morning and afternoon, and observe ewes. There are obvious signs in the behavior and reproductive organs of ewes when they are in estrus. When ewes are in estrus, they cry restlessly, shake their heads and tails, look around, urinate frequently, have a loss of appetite, and rumination and feeding time are significantly reduced. When in estrus, the vulva of the ewe is congested and swollen, and mucus secretion can be seen. The amount of mucus increases during the peak of estrus, and it is thin and transparent. Ewes with estrus are bred in time and records are kept. During this period, the corresponding feed is still used for feeding, and the feeding method remains unchanged. The results are shown in Table 3.

[0054] After the experiment, the pregnant ewes were tracked, the total number of lambs born was recorded during delivery, and the lambing rate of the ewes and the reproductive rate of the experimental flock were calculated.

[0055] Estrus rate = number of estrus ewes during the test period / total number of ewes in the flock × 100% Pregnancy rate = number of pregnant sheep during the test period / number of reference ewes during the test period × 100% Lambing rate = number of lambs born by ewes / number of ewes born × 100% Table 3

[0056] It can be seen from the above table that the feed for improving the estrus rate of ewes prepared in Examples 1-3 of the present invention significantly improved the estrus rate, pregnancy rate and lambing rate of ewes, and twin births occurred.

[0057] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a feed for improving the estrus rate of ewes, characterized in that: Epimedium, Morinda officinalis, Leonurus japonicus and leek seeds are fermented by selenium-rich yeast and manganese-rich yeast, and then precipitated with alcohol to obtain solid and liquid. The solid is a complex polysaccharide. A solid extractant is added to the liquid to obtain an extract, which is added to a basic diet together with the complex polysaccharide, stirred and mixed evenly to obtain a feed for improving the estrus rate of ewes.

2. The preparation method according to claim 1, characterized in that: The following steps are involved: S1. The selenium-enriched yeast and the manganese-enriched yeast were inoculated into a malt agar medium, activated and cultured to obtain a bacterial seed solution; S2. Wash, dry, and crush Epimedium, Morinda officinalis, Leonurus japonicus, and leek seeds, add them to water, add selenium-rich yeast and manganese-rich yeast seed solution, ferment and culture, filter, and retain the filtrate; S3. Add ethanol to the filtrate for precipitation, centrifuge, wash the solid, dry, and obtain a complex polysaccharide, and retain the centrifuge; S4. The ethanol is recovered from the centrifuged liquid, a solid extractant is added to the remaining liquid, adsorbed by stirring under an external magnetic field, separated by a magnet, the solid is added to ethanol, heated and stirred for desorption, the solid is separated by a magnet, recycled, the solvent is removed from the liquid under reduced pressure, and dried to obtain an extract; S5. Add soybean isoflavones, complex polysaccharides and extracts to the basic diet, stir and mix evenly, and prepare feed for improving the estrus rate of ewes.

3. The preparation method according to claim 2, wherein the activation culture conditions in step S1 are 28-32°C, 150-250r / min, activation culture 18-24h, and the bacterial content of the bacterial seed liquid is 10 8 -10 9 cfu / mL.

4. The preparation method according to claim 2, wherein the mass ratio of epimedium, Morinda officinalis, Leonurus japonicus and leek seeds in step S2 is 10:4-6:2-3:3-5, the inoculation amounts of the selenium-enriched yeast and manganese-enriched yeast seed solutions are 1-2 v / v% and 0.5-1.5 v / v%, respectively, and the fermentation and culture conditions are 28-32°C, 150-250 r / min, and the fermentation and culture is 48-72 h.

5. The preparation method according to claim 2, wherein in step S3, ethanol is added until the ethanol content of the system is 80-85wt%, and the precipitation time is 1-2h.

6. According to the preparation method of claim 2, the preparation method of the solid extractant in step S4 is as follows: T1. Add silica microspheres to water, add ferric chloride and ferrous chloride, add ammonia water under inert gas protection, heat and stir to react, centrifuge, wash, dry, and calcine to obtain magnetic silica microspheres; T2. adding magnetic silica microspheres, phytic acid, β-cyclodextrin, and potassium dihydrogen phosphate into water, performing hydrothermal reaction, separating with a magnet, washing, and drying to obtain a magnetic silica-phytic acid-cyclodextrin composite material; T3. Add the magnetic silica-phytic acid-cyclodextrin composite material into Tris-HCl solution, add dopamine hydrochloride, heat and stir to react, separate with a magnet, wash, and dry to obtain a solid extractant.

7. The preparation method according to claim 6, wherein the particle size of the silica microspheres in step T1 is 200-500 nm, the mass ratio of the silica microspheres, ferric chloride, ferrous chloride and ammonia water is 10-12:3.24:1.26:4-6, the temperature of the heating and stirring reaction is 75-85 ° C, the time is 3-5h, the temperature of the calcination is 400-500 ° C, the time is 0.5-1.5h; the magnetic silica microspheres in step T2 are The mass ratio of magnetic silica, phytic acid, β-cyclodextrin and potassium dihydrogen phosphate is 10:1-2:4-6:1-2, the temperature of the hydrothermal reaction is 140-160°C, and the time is 9-11h; the pH value of the Tris-HCl solution in step T3 is 8.5-9.5, the mass ratio of the magnetic silica-phytic acid-cyclodextrin composite material and dopamine hydrochloride is 10:1-2, the temperature of the heating and stirring reaction is 40-50°C, and the time is 1-3h.

8. The preparation method according to claim 2, wherein the liquid-to-solid ratio of the liquid and solid extractants in step S4 is 5-10 mL:1 g, the strength of the external magnetic field is 0.05-0.1 T, the stirring adsorption time is 30-50 min, the heating temperature is 40-50° C., and the stirring desorption time is 10-20 min.

9. The preparation method according to claim 2, wherein the basic diet in step S5 is mixed by the following raw materials in parts by weight: 10-20 parts of wheat straw, 4-6 parts of alfalfa, 20-25 parts of refined grain supplement, 20-30 parts of straw, and 15-20 parts of sweet potato starch; the refined grain supplement is mixed by the following raw materials in parts by weight: 10-30 parts of distiller's grains, 5-15 parts of soybean meal, 3-7 parts of vegetable oil, 1-3 parts of molasses, 1-2 parts of salt, 0.5-1.5 parts of calcium phosphate, 1-2 parts of vitamin C, 0.5-1.5 parts of vitamin A, and 0.3-0.5 parts of vitamin E; and the mass ratio of the basic diet, soybean isoflavones, complex polysaccharides, and extract is 100:0.5-1:8-10:10-12.

10. A feed for improving the estrus rate of ewes prepared by the preparation method according to any one of claims 1 to 9.

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