Coated cobalamin additive as well as preparation method and application thereof

Abstract

The invention provides a coated cobalamin additive as well as a preparation method and application thereof, and belongs to the technical field of animal additives. According to the coated cobalamin additive provided by the invention, the cobalamin is reasonably coated, so that the cobalamin has certain stability in rumen, and most of the cobalamin is ensured to be quickly released in small intestines, so that the absorption amount of an organism is increased. The coated cobalamin additive can reduce the waste of cobalamin in rumen and improve the absorption and utilization rate of cobalamin in small intestines. The feed cost can be reduced, and the utilization efficiency of ruminants on other nutritional ingredients in the feed can be improved.

Description

Technical Field

[0001] The invention belongs to the technical field of animal additives, and particularly relates to a coated cobalamin additive and a preparation method and application thereof. Background Art

[0002] Cobalamin (vitamin B12) is an important nutrient for the growth, development and production performance of ruminants. Cobalt is an essential active ingredient in vitamin B12. Ruminants synthesize very little of it themselves, and most of it needs to be ingested through feed or additives. However, cobalamin is mainly absorbed in the small intestine, and the absorption rate of cobalamin and its analogues synthesized in the rumen is low, at only 1% to 3%. If cobalamin is fed directly, 63.9% of the cobalamin will be degraded or destroyed by rumen microorganisms in the rumen, resulting in a reduction in its absorbable amount in the small intestine. Summary of the Invention

[0003] In view of this, the object of the present invention is to provide a coated cobalamin additive, which, by reasonably coating the cobalamin, makes it have a certain stability in the rumen, ensures that most of it is quickly released in the small intestine, thereby increasing the body's absorption.

[0004] The invention provides a coated cobalamin additive, which comprises the following components in parts by mass: 0.18-0.22 parts of cobalamin, 26.24-32.34 parts of silicon dioxide, 0.08-0.14 parts of lipase, 28.7-32.7 parts of palm fat, 22.7-26.7 parts of coconut oil and 12-18 parts of calcium stearate.

[0005] Preferably, the following components are included in parts by mass: 0.19-0.21 parts of cobalamin, 28.26-30.28 parts of silicon dioxide, 0.10-0.12 parts of lipase, 29.7-30.7 parts of palm fat, 23.7-24.7 parts of coconut oil and 14-17 parts of calcium stearate.

[0006] Preferably, the following components are included in parts by mass: 0.2 parts of cobalamin, 29.3 parts of silicon dioxide, 0.1 parts of lipase, 30.7 parts of palm fat, 24.7 parts of coconut oil and 15 parts of calcium stearate.

[0007] The present invention provides a method for preparing the coated cobalamin additive, comprising the following steps:

[0008] mixing cobalamin, lipase and a portion of silicon dioxide to obtain a first mixed material;

[0009] Mixing the first mixed material and the remaining amount of silicon dioxide again to obtain a second mixed material;

[0010] Mixing and granulating the heated coconut oil, a portion of the heated palm fat and the second mixed material to obtain granules;

[0011] The granules were coated with the remaining amount of heated palm fat and heated calcium stearate to obtain a coated cobalamin additive.

[0012] Preferably, after the mixing and granulation, the particles are collected through a sieve with an aperture of 0.8 mm to 1.25 mm.

[0013] Preferably, the heating temperature of the palm fat or calcium stearate is 55-65°C.

[0014] The present invention provides an animal feed comprising the coated cobalamin additive or the coated cobalamin additive prepared by the preparation method.

[0015] Preferably, the added amount of the coated cobalamin additive is 0.6 to 0.9 mg / kg based on the mass of cobalamin relative to the total mass of the animal feed.

[0016] The present invention provides the coated cobalamin additive, the coated cobalamin additive prepared by the preparation method or the animal feed in ruminant breeding.

[0017] Preferably, the animal husbandry includes at least one of the following: promoting the lactation performance of ruminants, promoting the growth of ruminants, improving the feed utilization rate of ruminants, improving the absorption of cobalamin by ruminants, improving the digestibility of nutrients in ruminants and improving the energy metabolism state of ruminants.

[0018] The present invention provides a coated cobalamin additive comprising the following components by weight: 0.18-0.22 parts cobalamin, 26.24-32.34 parts silicon dioxide, 0.08-0.14 parts lipase, 28.7-32.7 parts palm fat, 22.7-26.7 parts coconut oil, and 12-18 parts calcium stearate. The present invention utilizes the adsorption effect of silicon dioxide to adsorb the lipase and cobalamin near the silicon dioxide, while coconut oil is used to adjust the melting point. The palm fat acts as a binder and reduces the degradation rate of rumen microorganisms, while the calcium stearate also reduces the degradation rate of rumen microorganisms. The lipase digests fat in the small intestine and promotes the release of cobalamin in the small intestine. Experimental results show that the 24-hour degradation rate of the coated cobalamin additive in the rumen is 18.5% to 27.4%, with Example 5 having the lowest 24-hour rumen degradation rate (18.5%). The release rate of the coated cobalamin additive prepared by the present invention in the small intestine is 55.7% to 70.5%, among which the coated cobalamin additive prepared in Example 5 has the highest release rate in the small intestine (70.5%). At the same time, experiments show that the coated cobalamin additive can not only significantly improve the production performance of dairy cows but also promote the growth performance of beef cattle. At the same time, the addition of coated cobalamin can increase the pH value of rumen fluid, improve rumen fermentation and rumen microbial flora, help improve the digestion and metabolism ability of ruminants to feed, and ultimately achieve the purpose of promoting lactation performance. Adding coated cobalamin additives to feed with a high proportion of concentrate can effectively solve the problem of rumen acidosis caused by long-term feeding of high-concentrate feed. By increasing the absorption of vitamin B12, the pyruvate metabolic pathway is regulated, the activity of key enzymes in lactate synthesis is reduced, and the production of lactic acid is reduced, thereby increasing the pH of rumen fluid and improving rumen fermentation. Coated cobalamin additives can reduce the waste of cobalamin in the rumen and improve its absorption and utilization rate in the small intestine. This not only helps reduce feed costs but also improves the efficiency with which ruminants utilize other nutrients in their feed. DETAILED DESCRIPTION

[0019] The invention provides a coated cobalamin additive, which comprises the following components in parts by mass: 0.18-0.22 parts of cobalamin, 26.24-32.34 parts of silicon dioxide, 0.08-0.14 parts of lipase, 28.7-32.7 parts of palm fat, 22.7-26.7 parts of coconut oil and 12-18 parts of calcium stearate.

[0020] In the present invention, the coated cobalamin additive preferably includes the following components in parts by mass: 0.19-0.21 parts of cobalamin, 28.26-30.28 parts of silicon dioxide, 0.10-0.12 parts of lipase, 29.7-30.7 parts of palm fat, 23.7-24.7 parts of coconut oil and 14-17 parts of calcium stearate.

[0021] In the present invention, the coated cobalamin additive preferably includes the following components in parts by mass: 0.18 parts of cobalamin, 32.34 parts of silicon dioxide, 0.08 parts of lipase, 32.7 parts of palm fat and 22.7 parts of coconut oil, and 12 parts of calcium stearate. The coated cobalamin additive preferably includes the following components in parts by mass: 0.18 parts of cobalamin, 29.32 parts of silicon dioxide, 0.1 parts of lipase, 32.7 parts of palm fat and 22.7 parts of coconut oil, and 15 parts of calcium stearate. The coated cobalamin additive preferably includes the following components in parts by mass: 0.18 parts of cobalamin, 27.3 parts of silicon dioxide, 0.12 parts of lipase, 32.7 parts of palm fat and 22.7 parts of coconut oil, and 17 parts of calcium stearate. The coated cobalamin additive preferably includes the following components in parts by mass: 0.2 parts of cobalamin, 32.32 parts of silicon dioxide, 0.08 parts of lipase, 30.7 parts of palm fat and 24.7 parts of coconut oil, and 12 parts of calcium stearate. The coated cobalamin additive preferably includes the following components in parts by mass: 0.2 parts of cobalamin, 29.3 parts of silicon dioxide, 0.1 parts of lipase, 30.7 parts of palm fat, 24.7 parts of coconut oil, and 15 parts of calcium stearate. The coated cobalamin additive preferably includes the following components in parts by mass: 0.2 parts of cobalamin, 27.28 parts of silicon dioxide, 0.12 parts of lipase, 30.7 parts of palm fat and 24.7 parts of coconut oil, and 17 parts of calcium stearate. The coated cobalamin additive preferably includes the following components in parts by mass: 0.22 parts of cobalamin, 32.3 parts of silicon dioxide, 0.8 parts of lipase, 28.7 parts of palm fat and 26.7 parts of coconut oil, and 12 parts of calcium stearate. The coated cobalamin additive preferably includes the following components in parts by mass: 0.22 parts of cobalamin, 30.28 parts of silicon dioxide, 0.1 parts of lipase, 28.7 parts of palm fat and 26.7 parts of coconut oil, and 14 parts of calcium stearate. The coated cobalamin additive preferably includes the following components in parts by mass: 0.22 parts of cobalamin, 28.26 parts of silicon dioxide, 0.12 parts of lipase, 28.7 parts of palm fat and 26.7 parts of coconut oil, and 16 parts of calcium stearate. The coated cobalamin additive preferably comprises the following components by weight: 0.22 parts cobalamin, 27.24 parts silicon dioxide, 0.14 parts lipase, 28.7 parts palm fat, 26.7 parts coconut oil, and 18 parts calcium stearate. The silicon dioxide is preferably a 400-mesh fine powder. The silicon dioxide acts as an adsorbent; the lipase digests fat in the small intestine, promoting the release of cobalamin; the palm fat acts as a binder and slows the degradation of vitamins in the rumen; the coconut oil adjusts the melting point; and the calcium stearate slows the degradation of vitamins in the rumen.

[0022] In the present invention, the coated cobalamin additive is a spherical particle with a two-layer structure, divided into a core and an outer layer from the inside out. The core is a mixture of cobalamin, lipase, silicon dioxide, palm fat, and coconut oil. The outer layer is a mixture of palm fat and calcium stearate. Because the outer layer of the coated cobalamin additive encapsulates the palm fat and calcium stearate, rumen microorganisms first digest the outer layer composed of palm fat and calcium stearate in the rumen. The outer layer effectively blocks the degradation of the cobalamin in the coated cobalamin additive by microorganisms, protecting the cobalamin from degradation. Only a portion of the cobalamin is released in the rumen to promote the growth and reproduction of rumen microorganisms. When the coated cobalamin additive reaches the small intestine, the coconut oil adjusts the melting point of the core, causing it to gradually decompose. At the same time, the lipase reaches suitable enzymatic hydrolysis conditions in the small intestine, thereby degrading the palm fat that acts as a binder in the core, thereby fully releasing the cobalamin from the core and improving the absorption and utilization efficiency of the small intestine.

[0023] The present invention provides a method for preparing the coated cobalamin additive, comprising the following steps:

[0024] mixing cobalamin, lipase and a portion of silicon dioxide to obtain a first mixed material;

[0025] Mixing the first mixed material and the remaining amount of silicon dioxide again to obtain a second mixed material;

[0026] Mixing and granulating the heated coconut oil, a portion of the heated palm fat and the second mixed material to obtain granules;

[0027] The granules were coated with the remaining amount of heated palm fat and heated calcium stearate to obtain a coated cobalamin additive.

[0028] In the present invention, cobalamin, lipase and a portion of silicon dioxide are mixed to obtain a first mixed material.

[0029] In the present invention, the mixing is preferably carried out in a waist drum mixer. The partial amount of silicon dioxide accounts for 8% to 12% of the total mass of silicon dioxide, and may be 10%.

[0030] After obtaining the first mixed material, the present invention mixes the first mixed material and the remaining amount of silicon dioxide again to obtain a second mixed material.

[0031] In the present invention, the remixing is preferably carried out in a V-type mixer. After the mixing and granulation, the mixture is passed through a sieve with a pore size of 1.25 mm, the undersize is collected, and the mixture is passed through a sieve with a pore size of 0.8 mm, and the particles on the oversize are collected.

[0032] After obtaining the second mixed material, the present invention mixes the heated coconut oil, a portion of the heated palm fat and the second mixed material to form granules.

[0033] In the present invention, the partial amount of palm fat accounts for 84% to 96% of the total mass of palm fat. The heating temperature of the palm fat or calcium stearate is preferably 55 to 65° C., and can be 60° C. The granulation is preferably completed in a rotary granulator.

[0034] After the granules are obtained, the outer layer of the granules is coated with the remaining amount of heated palm fat and heated calcium stearate to obtain the coated cobalamin additive.

[0035] In the present invention, the heating temperature of the palm fat or calcium stearate is preferably 55-65°C, and can be 60°C.

[0036] The present invention provides an animal feed comprising the coated cobalamin additive or the coated cobalamin additive prepared by the preparation method.

[0037] In the present invention, the amount of the coated cobalamin additive added is preferably 0.6-0.9 mg / kg of the total mass of the animal feed based on the mass of cobalamin, and can be 0.7-0.8 mg / kg. The animal feed preferably also includes a basal diet. The present invention has no special restrictions on the type of prime basal diet, and the basal diet well known in the art can be used. The present invention has no special restrictions on the composition of the basal diet, and the basal diet for conventional ruminants such as cattle can be used. In a specific embodiment, the basal diet is composed of: 25-26% corn silage, 11.4-13.2% alfalfa hay, 11.8-12.6% oat hay, 25.4-25.8% corn, 5.5-6.2% wheat bran, 9.2-9.8% soybean meal, 2.4-2.5% rapeseed meal, 4.6-5.0% cottonseed meal, 0.5% calcium carbonate, 0.5% salt, 0.3% calcium hydrogen phosphate, and 0.5% premix. In a specific embodiment, the ruminant feed is fed twice daily, with each feeding amount being 12-17 kg, specifically 15 kg, with free access to water. In a specific embodiment, the corn may be steam-flaked corn. In a specific embodiment, the premix includes minerals and vitamins; in a specific embodiment, the minerals include iron, copper, manganese, zinc, iodine, selenium, and cobalt, and the vitamins include vitamin A, vitamin D, and vitamin E.

[0038] The present invention provides the coated cobalamin additive, the coated cobalamin additive prepared by the preparation method or the animal feed in ruminant breeding.

[0039] In the present invention, the animal husbandry preferably includes at least one of the following: promoting the lactation performance of ruminants, promoting the growth of ruminants, improving the feed utilization rate of ruminants, improving the absorption of cobalamin by ruminants, improving the digestibility of nutrients in ruminants and improving the energy metabolism state of ruminants. Improving the lactation performance of ruminants; the lactation performance includes milk production, milk fat corrected milk volume, milk fat content and milk protein content. Improving the energy metabolism state of ruminants; includes increasing the glucose, total protein, albumin, insulin and folic acid levels in the blood of dairy cows, and reducing the β-hydroxybutyric acid content in the blood of cows. The ruminants also preferably include meat-type ruminants and / or tolerance-producing ruminants. The ruminants include cattle or sheep.

[0040] The following is a detailed description of a coated cobalamin additive provided by the present invention, its preparation method and application, in conjunction with the examples. However, these examples should not be construed as limiting the scope of protection of the present invention.

[0041] Example 1

[0042] A preparation method of coated cobalamin additive

[0043] 1. Weigh 180g of feed-grade cobalamin, 80g of lipase, and 3kg of silicon dioxide (400 mesh).

[0044] 2. Mix the materials in step 1 evenly in a waist drum mixer.

[0045] 3. Add 29.34 kg of silicon dioxide (400 mesh) to the mixed material obtained in step 2 and mix evenly in a V-type mixer to obtain a mixed material;

[0046] 4. Heat 27.7 kg of palm fat and 22.7 kg of coconut oil to 60°C, mix evenly with the mixture in step 3, pass through a 1.25 mm sieve, collect the undersize, pass through a 0.8 mm sieve, collect the particles on the sieve, and perform rotary granulation in a rotary granulator mixer;

[0047] 5. Place the granules prepared in step 4 into a rotary granulator and coat the outer layer of the granules prepared in step 4 with 5 kg of palm fat heated to 60 degrees and 12 kg of calcium stearate.

[0048] Example 2

[0049] A preparation method of coated cobalamin additive

[0050] Compared with the preparation method of Example 1, the difference is that the content of each component is as follows: 180g feed-grade cobalamin, 100g lipase, 3kg silicon dioxide (premixed in step 1), 26.32kg silicon dioxide (mixed again in step 3), 27.7kg palm fat and 22.7kg coconut oil, 5kg 60-degree palm fat and 15kg calcium stearate.

[0051] Example 3

[0052] A preparation method of coated cobalamin additive

[0053] Compared with the preparation method of Example 1, the difference is that the content of each component is as follows: 180g feed-grade cobalamin, 120g lipase, 3kg silicon dioxide (premixed in step 1), 24.3kg silicon dioxide (mixed again in step 3), 27.7kg palm fat, 22.7kg coconut oil, 5kg 60-proof palm fat and 17kg calcium stearate.

[0054] Example 4

[0055] A preparation method of coated cobalamin additive

[0056] Compared with the preparation method of Example 1, the difference is that the content of each component is as follows: 200 g feed-grade cobalamin, 80 g lipase, 3 kg of silicon dioxide (premixed in step 1), 29.32 kg of silicon dioxide (400 mesh), 24.7 kg of palm fat, 24.7 kg of coconut oil, 6 kg of 60-degree palm fat, and 12 kg of calcium stearate.

[0057] Example 5

[0058] A preparation method of coated cobalamin additive

[0059] Compared with the preparation method of Example 1, the difference is that the content of each component is as follows: 200 g feed-grade cobalamin, 100 g lipase, 3 kg of silicon dioxide (premixed in step 1), 26.3 kg of silicon dioxide (mixed again in step 3), 24.7 kg of palm fat, 24.7 kg of coconut oil, 6 kg of 60-degree palm fat, and 15 kg of calcium stearate.

[0060] Example 6

[0061] A preparation method of coated cobalamin additive

[0062] Compared with the preparation method of Example 1, the difference is that the content of each component is as follows: 200 g feed-grade cobalamin, 120 g lipase, 3 kg of silicon dioxide (premixed in step 1), 24.28 kg of silicon dioxide (mixed again in step 3), 24.7 kg of palm fat, 24.7 kg of coconut oil, 6 kg of 60-degree palm fat, and 17 kg of calcium stearate.

[0063] Example 7

[0064] A preparation method of coated cobalamin additive

[0065] Compared with the preparation method of Example 1, the difference is that the content of each component is as follows: feed-grade cobalamin 220g, lipase 80g, silicon dioxide (premixed in step 1) 3kg, silicon dioxide (mixed again in step 3) 29.3kg, palm fat 21.7kg, coconut oil 26.7kg, 60-degree palm fat 7kg, and calcium stearate 12kg.

[0066] Example 8

[0067] A preparation method of coated cobalamin additive

[0068] Compared with the preparation method of Example 1, the difference is that the content of each component is as follows: feed-grade cobalamin 220g, lipase 100g, silicon dioxide (premixed in step 1) 3kg, silicon dioxide (mixed again in step 3) 27.28kg, palm fat 21.7kg, coconut oil 26.7kg, 60-proof palm fat 7kg, and calcium stearate 14kg.

[0069] Example 9

[0070] A preparation method of coated cobalamin additive

[0071] Compared with the preparation method of Example 1, the difference is that the content of each component is as follows: 220 g of premixed feed-grade cobalamin, 120 g of lipase, 3 kg of silicon dioxide (premixed in step 1), 25.26 kg of silicon dioxide (mixed again in step 3), 21.7 kg of palm fat, 26.7 kg of coconut oil, 7 kg of 60-proof palm fat, and 16 kg of calcium stearate.

[0072] Example 10

[0073] A preparation method of coated cobalamin additive

[0074] 1. Weigh 220g of feed-grade cobalamin, 140g of lipase, and 3kg of silicon dioxide (400 mesh);

[0075] 2. Mix the materials in step 1 evenly in a waist drum mixer;

[0076] 3. Add 23.24kg of silicon dioxide (400 mesh) to the mixed materials in step 2 and mix them evenly in a V-type mixer.

[0077] 4. Heat 21.7 kg of palm fat and 26.7 kg of coconut oil to 60 degrees, mix evenly with 3, pass through a 0.8 mm sieve, and perform rotary granulation in a rotary granulator mixer;

[0078] 5. Place the granules prepared in step 4 into a rotary granulator and coat the granules prepared in step 2.4 with 18 kg of calcium stearate using 7 kg of palm fat heated to 60 degrees.

[0079] Example 11

[0080] Comparative test of the degradation rate in the rumen and the release rate in the small intestine of the coated cobalamin additives prepared in Examples 1-10

[0081] Twelve dairy bulls (weighing 580 ± 11.2 kg) fitted with permanent rumen and duodenal cannulas were randomly divided into four groups of three. To eliminate inter-animal variability, a 4×4 Latin square design was used, with four phases. Each phase consisted of a 10-day pre-feeding period and a 5-day feeding period.

[0082] The basal diet consisted of 25% corn silage, 13.2% alfalfa hay, 11.8% oat hay, 25.8% corn, 5.5% bran, 9.8% soybean meal, 2.5% rapeseed meal, 4.6% cottonseed meal, 0.5% calcium carbonate, 0.5% salt, 0.3% calcium hydrogen phosphate, and 0.5% premix. The cattle were fed 15 kg of the complete mixed ration twice daily (7:00 and 19:00) in a single trough. Water was available ad libitum.

[0083] For each trial, 5g of each coated cobalamin from Examples 1-10 was accurately weighed and placed into nylon bags (5cm x 8cm) of known mass. The bags were tied with nylon string and placed 50cm from the rumen sac after the morning feed on the first day of the formal period. Eight bags were placed per cow, and four bags were removed at 12 and 24 hours. The two bags removed from the rumen were immediately rinsed with water until the water was completely clear and then dried at 65°C to constant weight. The cobalamin content before and after degradation was determined using the method described in GB / T 17819-2017, Determination of Vitamin B12 in Additive Premixes (High-Performance Liquid Chromatography). Two additional bags were placed into the small intestine of the cow through the duodenal fistula. The nylon bags were carefully collected from the cow feces, immediately rinsed with water until the water was completely clear, and then dried at 65°C to constant weight. The cobalamin content in the residue was determined using the method described in GB / T 17819-2017. The rumen degradation rate and small intestinal release rate were calculated. The calculation results are shown in Table 1.

[0084] Rumen degradation rate = (mass of coated cobalamin before degradation × cobalamin content before degradation - mass of coated cobalamin after degradation × cobalamin content after degradation) / (mass of coated cobalamin before degradation × cobalamin content before degradation) × 100% Formula I;

[0085] Small intestine release rate = (mass of coated cobalamin before degradation × cobalamin content before degradation - mass of coated cobalamin residue in the nylon bag collected in feces × cobalamin content in the residue) / (mass of coated cobalamin before degradation × cobalamin content before degradation) × 100% Formula II.

[0086] Table 1 Rumen degradation rate and small intestine release rate of cobalamin

[0087]

[0088] Note: Different letters in the same row indicate significant differences (P<0.05).

[0089] As shown in Table 1, the 12-hour rumen degradation rates of the coated cobalamin additives prepared in Examples 1-10 ranged from 10.9% to 18.6%, with Example 5 having the lowest 12-hour rumen degradation rate (10.9%), significantly lower than the other examples except Example 6. The 24-hour rumen degradation rates of the coated cobalamin additives in Examples 1-10 ranged from 18.5% to 27.4%, with Example 5 having the lowest 24-hour rumen degradation rate (18.5%), significantly lower than the other examples. The small intestine release rates of the coated cobalamin additives prepared in Examples 1-10 ranged from 55.7% to 70.5%, with Example 5 having the highest small intestine release rate (70.5%), significantly higher than the other examples. This indicates that the coated cobalamin additive in Example 5 is more effective in crossing the rumen and entering the small intestine than the cobalamin in the other examples.

[0090] Example 12

[0091] 1 Materials and Methods

[0092] 1.1 Experimental design and feeding management

[0093] In this experiment, 40 Holstein lactating dairy cows (parity of 2.3±0.78, body weight of 626±12.1kg, lactation days of 45.8±2.6d, daily milk production of 33.6±3.2kg) were randomly divided into 4 groups, namely a control group, a low-level coated cobalamin group (6mg cobalamin / d), a medium-level coated cobalamin group (12mg cobalamin / d) and a high-level coated cobalamin group (18mg cobalamin / d). The experiment lasted for 70 days, including a 10-day pre-feeding period and a 60-day formal test period. The concentrate-to-roughage ratio was 50:50. The specific composition and nutritional level are shown in Table 2. During the experiment, the experimental cows were raised in a single pen, with free access to food and water, and fed at 07:00, 13:00 and 19:00 every day. The coated cobalamin used in the experiment was prepared according to Example 5 of the method of the present invention.

[0094] Table 2 Nutrient composition and chemical composition of basal diet (dry matter basis)

[0095] composition content(%) chemical composition content(%) Corn silage 26.0 organic matter 95.36 Alfalfa hay 11.2 crude protein 16.03 Oat hay 12.8 crude fat 5.76 Steam-flaked corn 25.6 Neutral detergent fiber 37.83 wheat bran 6.0 Acid detergent fiber 18.24 soybean meal 9.0 calcium 0.71 rapeseed meal 2.6 phosphorus 0.42 cottonseed cake 5.0 calcium carbonate 0.5 salt 0.5 Calcium hydrogen phosphate 0.3 Mineral and vitamin premix1 0.5

[0096] *Each kg of premix contains: Iron 20,000 mg, Copper 1,600 mg, Manganese 8,000 mg, Zinc 7,500 mg, Iodine 1,20 mg, Cobalt 20 mg, Vitamin A 820,000 IU, Vitamin D 300,000 IU and Vitamin E 10,000 IU.

[0097] 1.2 Measurement items and sample collection and analysis

[0098] 1.2.1 Feed intake measurement and feed sample collection and measurement

[0099] During the experiment, the amount of feed fed to the cows and the amount of feed leftover were recorded daily. Diet samples were collected and weighed every 10 days. Diets were placed in trays and dried at 60-65°C. Samples were then ground and analyzed for dry matter (DM), organic matter (OM), protein (CP), crude fat (EE), neutral detergent fiber (NDF), and acid detergent fiber (ADF) using conventional methods.

[0100] 1.2.2 Milk production determination and milk sample collection and analysis

[0101] During the experiment, milk production was recorded daily, and milk samples were collected every 10 days. 50 mL of the total milk sample was taken at 7:00, 1:00, and 19:00. Potassium dichromate (0.6 mg / mL) was added to the milk samples and the milk component content was determined using an infrared dairy analyzer. The milk component content was calculated using a weighted average algorithm. The lactocobalamin content was determined using high-performance liquid chromatography.

[0102] 1.2.3 Fecal sample collection and analysis

[0103] Feces were collected rectally. From day 68 to day 70 of the experiment, 5 experimental cows were randomly selected from each group. Feces were collected once every day at 7:00, 13:00 and 19:00 respectively. 300 g of feces were collected from each cow at a time, and 10% tartaric acid solution (1 / 4 of the feces weight) was added for nitrogen fixation. The fecal samples collected from each experimental cow for 3 days were thoroughly mixed to prepare air-dried fecal samples. The dry matter (DM), organic matter (OM), protein (CP), crude fat (EE), neutral detergent fiber (NDF) and acid detergent fiber (ADF) in the fecal samples were determined.

[0104] 1.2.4 Collection of rumen fluid and determination of fermentation parameters

[0105] On the 70th day of the experiment, rumen fluid was collected via gastric tube 2 hours after the morning feed. To avoid contamination with saliva, the first portion collected was discarded. Approximately 200 mL of rumen fluid sample was collected from each cow, the pH was immediately measured using a PHS-2C pH meter, filtered through four layers of gauze, and stored frozen at -40°C. Volatile fatty acids (VFA) and ammonia nitrogen (Ammonia N) were determined using gas chromatography.

[0106] 1.2.5 Collection and determination of blood samples

[0107] On days 30 and 60 of the formal trial period, 15 mL of blood was collected from the caudal vein of each cow in a blood collection tube after morning milking and before feeding. The samples were centrifuged at 4°C and 2000 × g for 15 minutes. The supernatant serum was extracted and stored at -20°C for blood parameter determination. Serum albumin (AlB), glucose (GLU), total protein (TP), and triglyceride (TG) levels were measured using a UV-visible spectrophotometer. Specific methods were described in the kit instructions. Serum CA, urea nitrogen (BUN), β-hydroxybutyrate (BHB), and insulin (INS) levels were measured using a microplate reader, following the instructions in the kit instructions.

[0108] 1.3 Data processing and statistical analysis

[0109] The data were analyzed using SAS 9.0 statistical analysis software for variance analysis model.

[0110] 2 Results

[0111] 2.1 Dry matter intake, lactation performance, and feed efficiency

[0112] As shown in Table 3, with the linear increase of coated cobalamin, the dry matter intake of dairy cows decreased linearly, and the high-level coated cobalamin group was significantly lower than the low-level coated cobalamin, medium-level coated cobalamin and control group (P<0.05); the actual milk production increased quadratically (P<0.05), and the low-level coated cobalamin and medium-level coated cobalamin groups were significantly higher than the high-level coated cobalamin and control group; the milk fat corrected milk and milk fat yield increased linearly (P<0.05), and the medium-level coated cobalamin group and high-level coated cobalamin group were significantly higher than the control group (P<0.05); the milk protein yield increased quadratically (P<0.05), and the medium-level coated cobalamin group was significantly increased compared with the control group (P<0.05), There was no significant change between the low-level coated cobalamin and high-level coated cobalamin groups (P>0.05); lactose production increased quadratically (P<0.05), and compared with the control group, the low-level coated cobalamin and medium-level coated cobalamin groups increased significantly (P<0.05), while the high-level coated cobalamin group showed no significant change (P>0.05); milk fat rate increased linearly, and compared with the control group, the high-level coated cobalamin group increased significantly, while the low-level coated cobalamin and medium-level coated cobalamin groups showed no significant change (P>0.05); there was no significant change in milk protein and lactose rates (P>0.05); feed efficiency increased linearly (P<0.05), and the coated cobalamin supplemented group was significantly higher than the control group (P<0.05).

[0113] Table 3 Effects of supplementation with coated cobalamin on lactation performance of lactating dairy cows

[0114]

[0115] Note: Different letters in the same row indicate significant differences (P<0.05).

[0116] 2.2 Nutrient digestibility

[0117] As shown in Table 4, with the linear increase of coated cobalamin, the digestibility of dry matter, organic matter, crude protein, crude fat, neutral detergent fiber and acid detergent fiber increased linearly (P<0.05). Compared with the control group, the digestibility of dry matter, organic matter, crude protein and crude fat in the medium-level coated cobalamin group and the high-level coated cobalamin group increased the most, followed by the low-level coated cobalamin group (P<0.05); compared with the control group and the low-level coated cobalamin group, the digestibility of neutral detergent fiber in the high-level coated cobalamin and medium-level coated cobalamin groups was significantly increased (P<0.05); compared with the control group, the addition of coated cobalamin significantly increased the digestibility of acid detergent fiber (P<0.05).

[0118] Table 4 Effects of supplementation with coated cobalamin on nutrient digestibility in lactating dairy cows

[0119]

[0120] Note: Different letters in the same row indicate significant differences (P<0.05).

[0121] 2.3 Rumen fermentation

[0122] As shown in Table 5, the supplementation of gradient-coated cobalamin in the diet had no significant effect on rumen pH (P>0.05); the total volatile acid concentration increased quadratically (P<0.05), and compared with the control group, the medium-level coated cobalamin group had the largest increase, followed by the low-level coated cobalamin group and the high-level coated cobalamin group (P<0.05); the acetic acid molar ratio and ammonium nitrogen concentration decreased linearly, and compared with the control group, the medium-level coated cobalamin group was significantly reduced (P<0.05), and the low-level coated cobalamin group was significantly reduced (P<0.05). There was no significant change in the content of propionic acid in the coated cobalamin group and the high level of coated cobalamin group (P>0.05); compared with the control group, the molar ratio of propionic acid in the coated cobalamin group increased linearly (P<0.05); the molar ratio of butyric acid, valeric acid, isobutyric acid and isovaleric acid did not change significantly (P>0.05); the ratio of acetic acid / propionic acid decreased linearly (P<0.05), and was significantly reduced in the medium level and high level of coated cobalamin groups compared with the control group (P<0.05), and there was no significant change in the low level of coated cobalamin group (P>0.05).

[0123] Table 5 Effects of supplementation with coated cobalamin on rumen fermentation in lactating dairy cows

[0124]

[0125]

[0126] Note: Different letters in the same row indicate significant differences (P<0.05).

[0127] 2.4 Blood indicators

[0128] As shown in Table 6, with the increase of the level of coated cobalamin in the diet, the concentrations of glucose, total protein, albumin, cobalamin and insulin in the blood increased linearly (P<0.05); compared with the control group, the glucose and albumin concentrations of the coated cobalamin group were significantly increased (P<0.05); the total protein content of the low-level coated cobalamin group and the medium-level coated cobalamin group was significantly higher than that of the control group (P<0.05), and there was no significant difference with the high-level coated cobalamin group (P>0.05); compared with the control group, the cobalamin and insulin contents of the medium-level coated cobalamin group and the high-level coated cobalamin group were significantly increased (P<0.05), and there was no significant difference with the low-level coated cobalamin group (P>0.05). As the level of coated cobalamin supplementation increased, the levels of non-esterified fatty acids and β-hydroxybutyrate in the blood decreased linearly (P<0.05). The levels of non-esterified fatty acids in the intermediate and high-cobalamin groups were significantly lower than those in the control group (P<0.05), but not significantly different from those in the low-cobalamin group (P>0.05). The β-hydroxybutyrate level in the high-cobalamin group was significantly lower than those in the control, low-cobalamin, and intermediate-cobalamin groups (P<0.05). Urea nitrogen and triglyceride levels did not change significantly with the addition of coated cobalamin to the diet (P>0.05).

[0129] Table 6 Effects of supplementation with coated cobalamin on blood parameters of lactating dairy cows

[0130]

[0131]

[0132] Note: Different letters in the same row indicate significant differences (P<0.05).

[0133] 3 Conclusion

[0134] Supplementation of 12 mg / d of coated cobalamin promoted the production of total volatile fatty acids (VFA) in the rumen, increased nutrient digestibility, and thus improved the lactation performance of dairy cows.

[0135] Example 13

[0136] 1 Materials and Methods

[0137] 1.1 Experimental design and feeding management

[0138] In this experiment, 60 Angus cattle (520.5±5.52kg, 395±10.2d of age) were randomly divided into 4 groups, namely a control group, a low-level coated cobalamin group (4mg cobalamin / d), a medium-level coated cobalamin group (6mg cobalamin / d) and a high-level coated cobalamin group (8mg cobalamin / d). The experiment lasted for 70 days, including a 10-day pre-feeding period and a 60-day formal test period. The concentrate-to-roughage ratio was 60:40, and the specific composition and nutritional level are shown in Table 7. During the experiment, the experimental cows were raised in a single pen, with free access to food and water, and fed at 07:00, 13:00 and 19:00 every day. The coated cobalamin used in the experiment was made according to Example 5 of the method of the present invention.

[0139] Table 7 Nutrient composition and chemical composition of basal diet (dry matter basis)

[0140]

[0141]

[0142] *Each kg of premix contains: Iron 20,000 mg, Copper 1,600 mg, Manganese 8,000 mg, Zinc 7,500 mg, Iodine 1,20 mg, Cobalt 20 mg, Vitamin A 820,000 IU, Vitamin D 300,000 IU and Vitamin E 10,000 IU.

[0143] 1.2 Measurement items and sample collection and analysis

[0144] 1.2.1 Feed intake measurement and feed sample collection and measurement

[0145] During the experiment, the amount of feed fed and leftover from the Angus cattle was recorded daily. Diet samples were collected every 10 days and weighed fresh. Diets were placed in trays and oven-dried at 60–65°C. Samples were then ground and analyzed for dry matter (DM), organic matter (OM), protein (CP), crude fat (EE), neutral detergent fiber (NDF), and acid detergent fiber (ADF) using conventional methods.

[0146] 1.2.2 Weight gain determination

[0147] At the beginning and end of the experimental period, the rats were weighed before morning feeding and the daily weight gain was calculated.

[0148] 1.2.3 Fecal sample collection and analysis

[0149] Feces were collected rectally. From day 68 to day 70 of the experiment, 5 experimental cows were randomly selected from each group. Feces were collected once every day at 7:00, 13:00 and 19:00 respectively. 300 g of feces were collected from each cow at a time, and 10% tartaric acid solution (1 / 4 of the feces weight) was added for nitrogen fixation. The fecal samples collected from each experimental cow for 3 days were thoroughly mixed to prepare air-dried fecal samples. The dry matter (DM), organic matter (OM), protein (CP), crude fat (EE), neutral detergent fiber (NDF) and acid detergent fiber (ADF) in the fecal samples were determined.

[0150] 1.2.4 Collection of rumen fluid and determination of fermentation parameters

[0151] On the 70th day of the experiment, rumen fluid was collected via gastric tube 2 hours after the morning feed. To avoid contamination with saliva, the first portion collected was discarded. Approximately 200 mL of rumen fluid sample was collected from each cow, the pH was immediately measured using a PHS-2C pH meter, filtered through four layers of gauze, and stored frozen at -40°C. Volatile fatty acids (VFA) and ammonia nitrogen (Ammonia N) were determined using gas chromatography.

[0152] 1.2.5 Collection and determination of blood samples

[0153] On days 30 and 60 of the formal trial period, 15 mL of blood was collected from the caudal vein of each cow in a blood collection tube after morning milking and before feeding. The samples were centrifuged at 4°C and 2000 × g for 15 minutes. The supernatant serum was extracted and stored at -20°C for blood parameter determination. Serum albumin (AlB), glucose (GLU), total protein (TP), and triglyceride (TG) levels were measured using a UV-visible spectrophotometer. Specific methods were described in the kit instructions. Serum CA, urea nitrogen (BUN), β-hydroxybutyrate (BHB), and insulin (INS) levels were measured using a microplate reader, following the instructions in the kit instructions.

[0154] 1.3 Data processing and statistical analysis

[0155] The data were analyzed using SAS 9.0 statistical analysis software for variance analysis model.

[0156] 2 Results

[0157] 2.1 Dry matter intake, weight gain performance, and feed efficiency

[0158] As shown in Table 8, with the linear increase of coated cobalamin, there was no significant change in dry matter intake and body weight of beef cattle. The average daily gain from 31 to 60 days and the entire period increased in a quadratic curve (P<0.05). The feed conversion rate from 0 to 30 days, 31 to 60 days, and the entire period decreased in a quadratic curve (P<0.05). The average daily gain from 31 to 60 days and the entire period in the coated cobalamin supplementation group was significantly higher than that in the control group (P<0.05), and the average daily gain in the medium-level coated cobalamin group was significantly higher than that in the low-level coated cobalamin group and the high-level coated cobalamin group (P<0.05). The feed conversion rate from 0 to 30 days, 31 to 60 days, and the entire period in the coated cobalamin supplementation group was significantly lower than that in the control group (P<0.05), and the feed conversion rate in the medium-level coated cobalamin group was significantly lower than that in the low-level coated cobalamin group and the high-level coated cobalamin group (P<0.05).

[0159] Table 8 Effects of supplementation with coated cobalamin on weight gain performance of beef cattle

[0160]

[0161]

[0162] Note: Different letters in the same row indicate significant differences (P<0.05).

[0163] 2.2 Nutrient digestibility

[0164] As shown in Table 9, with the linear increase of coated cobalamin, the digestibility of dry matter, organic matter, crude protein, crude fat, neutral detergent fiber and acid detergent fiber increased linearly (P<0.05). Compared with the control group, the digestibility of dry matter, organic matter, crude protein and crude fat in the medium-level coated cobalamin group and the high-level coated cobalamin group increased the most, followed by the low-level coated cobalamin group (P<0.05); compared with the control group and the low-level coated cobalamin group, the digestibility of neutral detergent fiber in the high-level coated cobalamin and medium-level coated cobalamin groups was significantly increased (P<0.05); compared with the control group, the addition of coated cobalamin significantly increased the digestibility of acid detergent fiber (P<0.05).

[0165] Table 9 Effects of supplementation with coated cobalamin on nutrient digestibility in beef cattle

[0166]

[0167] Note: Different letters in the same row indicate significant differences (P<0.05).

[0168] 2.3 Rumen fermentation

[0169] As shown in Table 10, with the increase of the level of coated cobalamin supplementation, the total volatile acid concentration in the rumen increased in a quadratic curve (P<0.05), the medium-level coated cobalamin group was significantly higher than that in the other groups (P<0.05), and the low-level coated cobalamin group was significantly higher than that in the control group and the high-level coated cobalamin group (P<0.05); the rumen pH and ammonium nitrogen concentration decreased in a quadratic curve (P<0.05), the pH in the medium-level coated cobalamin group was significantly lower than that in the other groups (P<0.05), and the NH3-N concentration in the coated cobalamin supplementation group was significantly lower than that in the control group (P<0.05); the propionic acid molar ratio increased linearly (P<0 0.05), the coated cobalamin group was significantly higher than the control group (P<0.05), and the high-level coated cobalamin group was significantly higher than the low-level coated cobalamin group and the medium-level coated cobalamin group (P<0.05); the acetic acid / propionic acid ratio decreased linearly (P<0.05), the medium-level coated cobalamin group and the high-level coated cobalamin group were significantly lower than the control group (P<0.05), and the high-level coated cobalamin group was significantly lower than the low-level coated cobalamin group and the medium-level coated cobalamin group (P<0.05); the molar ratios of acetic acid, butyric acid, valeric acid, isobutyric acid and isovaleric acid did not differ significantly among the different treatment groups (P>0.05).

[0170] Table 10 Effects of supplementation with coated cobalamin on rumen fermentation in beef cattle

[0171]

[0172] Note: Different letters in the same row indicate significant differences (P<0.05).

[0173] 2.4 Blood indicators

[0174] As shown in Table 11, with increasing dietary cobalamin levels, blood glucose, total protein, albumin, cobalamin, and insulin concentrations increased linearly (P<0.05). Compared with the control group, glucose and albumin concentrations in the cobalamin-enriched group were significantly increased (P<0.05). Total protein content was significantly higher in the low- and medium-level cobalamin-enriched groups than in the control group (P<0.05), but not significantly different from the high-level cobalamin-enriched group (P>0.05). Cobalamin and insulin contents were significantly higher in the medium- and high-level cobalamin-enriched groups than in the control group (P<0.05), but not significantly different from the low-level cobalamin-enriched group (P>0.05). Urea nitrogen and triglyceride levels did not change significantly with dietary cobalamin supplementation (P>0.05).

[0175] Table 11 Effects of supplementation with coated cobalamin on blood parameters of beef cattle

[0176]

[0177]

[0178] Note: Different letters in the same row indicate significant differences (P<0.05).

[0179] 3 Conclusion

[0180] Supplementation of 6 mg / d of coated cobalamin promoted the production of total volatile fatty acids (VFA) in the rumen of beef cattle, increased nutrient digestibility, and thus improved the weight gain performance of beef cattle.

[0181] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A coated cobalamin additive, characterized in that The invention comprises the following components in parts by mass: 0.18-0.22 parts of cobalamin, 26.24-32.34 parts of silicon dioxide, 0.08-0.14 parts of lipase, 28.7-32.7 parts of palm fat, 22.7-26.7 parts of coconut oil and 12-18 parts of calcium stearate.

2. The coated cobalamin additive according to claim 1, characterized in that The invention comprises the following components in parts by mass: 0.19-0.21 parts of cobalamin, 28.26-30.28 parts of silicon dioxide, 0.10-0.12 parts of lipase, 29.7-30.7 parts of palm fat, 23.7-24.7 parts of coconut oil and 14-17 parts of calcium stearate.

3. The coated cobalamin additive according to claim 1, characterized in that The invention comprises the following components in parts by mass: 0.2 parts of cobalamin, 29.3 parts of silicon dioxide, 0.1 parts of lipase, 30.7 parts of palm fat, 24.7 parts of coconut oil and 15 parts of calcium stearate.

4. The method for preparing the coated cobalamin additive according to any one of claims 1 to 3, characterized in that: The following steps are involved: mixing cobalamin, lipase and a portion of silicon dioxide to obtain a first mixed material; Mixing the first mixed material and the remaining amount of silicon dioxide again to obtain a second mixed material; Mixing and granulating the heated coconut oil, a portion of the heated palm fat and the second mixed material to obtain granules; The granules were coated with the remaining amount of heated palm fat and heated calcium stearate to obtain a coated cobalamin additive.

5. The preparation method according to claim 4, characterized in that: After the mixing and granulation, the granules are collected between sieves with apertures of 0.8 mm to 1.25 mm.

6. The preparation method according to claim 4, characterized in that: The heating temperature of the palm fat or calcium stearate is 55-65°C.

7. An animal feed, characterized in that The invention comprises the coated cobalamin additive according to any one of claims 1 to 3 or the coated cobalamin additive prepared by the preparation method according to any one of claims 4 to 6.

8. The animal feed according to claim 7, characterized in that The added amount of the coated cobalamin additive is 0.6-0.9 mg / kg based on the mass of cobalamin to the total mass of the animal feed.

9. Use of the coated cobalamin additive according to any one of claims 1 to 3, the coated cobalamin additive prepared by the preparation method according to any one of claims 4 to 6, or the animal feed according to claim 7 or 8 in ruminant breeding.

10. The use according to claim 9, characterized in that: The animal feeding comprises at least one of the following: promoting the lactation performance of ruminants, promoting the growth of ruminants, improving the feed utilization rate of ruminants, improving the absorption of cobalamin by ruminants, improving the digestibility of nutrients in ruminants and improving the energy metabolism state of ruminants.

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