A feed additive for increasing the content of branched-chain fatty acids in milk and a method for preparing the same

Abstract

The present application belongs to the technical field of feed additive, and particularly relates to a feed additive for increasing the content of branched-chain fatty acids in milk and a preparation method thereof.The feed additive for increasing the content of branched-chain fatty acids in milk comprises the following raw materials in parts by weight: 1-4 parts of leucine, 1-4 parts of isoleucine, 1-4 parts of valine, 1-2.5 parts of Prinsepia uniflora extract and 0.3-0.6 parts of sodium subtilisin lipopeptide.The feed additive for increasing the content of branched-chain fatty acids in milk is prepared by using different types of amino acids and Prinsepia uniflora extract and sodium subtilisin lipopeptide in combination, and can significantly increase the content of branched-chain fatty acids in milk, improve the quality of milk and the milk production performance of dairy cows after being used in the daily ration of dairy cows.The feed additive is safe in composition and has no toxic side effects.

Description

Technical Field

[0001] This invention belongs to the field of feed additive technology, and in particular relates to a feed additive for increasing the content of branched-chain fatty acids in milk and its preparation method. Background Technology

[0002] Branched-chain fatty acids (BCFAs) are a class of fatty acid components in milk fat with unique nutritional and health value, showing significant potential in regulating human health. In recent years, with the increasing demand for functional foods, dairy products rich in BCFAs have become a research hotspot. The BCFA content in milk directly affects its nutritional quality and added value; therefore, increasing the BCFA level in raw milk through nutritional regulation has become an important research direction in the livestock and food industries.

[0003] Currently, increasing BCFA content in milk mainly relies on regulating dairy cow diets. Existing technologies include supplementing with fiber-rich forage to indirectly increase BCFA synthesis by promoting rumen microbial fermentation and producing BCFA precursors. However, this method has limited and unstable effects, heavily influenced by forage variety, origin, and season, making it difficult to precisely increase BCFA content. Secondly, directly adding vegetable oils or oilseeds, such as sunflower seeds or flaxseed, can alter rumen fermentation patterns and mammary gland fat synthesis substrates, but it primarily affects the overall fatty acid composition, with little effect on specific components like BCFA. Excessive addition can lead to negative consequences such as rumen dysfunction and decreased milk fat percentage. Other methods include using some known rumen regulators, such as monensin ionocarriers. While these can alter the proportion of volatile fatty acids in the rumen to some extent, their effect on BCFA synthesis lacks specificity and is unclear. Furthermore, they pose safety and regulatory risks such as drug residues and resistance, limiting their application under the trend of green and healthy farming.

[0004] Therefore, it is of great significance to develop a feed additive that is low-cost, simple in process, can be mass-produced, and can continuously and significantly increase the BCFA content of milk and its preparation method. Summary of the Invention

[0005] To overcome the shortcomings of the prior art, the primary objective of this invention is to provide a feed additive that increases the content of branched-chain fatty acids in milk. This additive, through the combined use of multiple components, can significantly increase the total amount of branched-chain fatty acids and the proportion of beneficial components in milk without affecting the health and normal production performance of dairy cows, thereby enhancing the nutritional value of milk and the milk production performance of dairy cows; and meeting the market demand for functional dairy products.

[0006] Another object of the present invention is to provide a method for preparing a feed additive that increases the content of branched-chain fatty acids in milk.

[0007] The objective of this invention is achieved through the following technical solution: A feed additive for increasing the branched-chain fatty acid content in milk, comprising, by weight, the following ingredients: 1-4 parts leucine, 1-4 parts isoleucine, 1-4 parts valine, 1-2.5 parts *Symplocos edulis* extract, and 0.3-0.6 parts sodium subtilis lipopeptide.

[0008] Furthermore, a feed additive for increasing the branched-chain fatty acid content in milk comprises, by weight, the following ingredients: 3 parts leucine, 3 parts isoleucine, 3 parts valine, 1.5 parts *Symplocos edulis* extract, and 0.5 parts sodium subtilis lipopeptide.

[0009] Furthermore, the preparation method of the *Cynanchum paniculatum* extract is as follows: the leaves of *Cynanchum paniculatum* are dried, pulverized, added to an aqueous acetic acid solution, ultrasonically extracted, the extract is collected, concentrated under reduced pressure, and then freeze-dried to obtain the *Cynanchum paniculatum* extract.

[0010] Furthermore, the solid-liquid ratio of the leaves of the *Prickly ash* to the aqueous acetic acid solution is 1g:6-10mL.

[0011] Furthermore, the ultrasonic extraction temperature is 50-60℃; the time is 1-3h.

[0012] Furthermore, the acetic acid in the aqueous acetic acid solution has a mass concentration of 5-8%.

[0013] A method for preparing a feed additive that increases the content of branched-chain fatty acids in milk includes the following steps: leucine, isoleucine, valine, *Prickly ash* extract, and *Bacillus subtilis* lipopeptide sodium are mixed evenly according to the specified ratio to obtain the additive.

[0014] The present invention has the following advantages over the prior art: 1. This invention provides a feed additive for increasing the branched-chain fatty acid content in milk. It uses different types of amino acids, extracts from *Symplocos edulis*, and sodium subtilis lipopeptide in combination. When applied to the diet of dairy cows, it can significantly increase the branched-chain fatty acid content in milk, improve milk quality, and enhance the milk production performance of dairy cows.

[0015] 2. This invention provides a feed additive that increases the branched-chain fatty acid content in milk. The feed additive is safe, has no toxic side effects, and meets the market demand for functional dairy products.

[0016] 3. This invention provides a method for preparing a feed additive that increases the content of branched-chain fatty acids in milk. This method is simple to operate and is suitable for addition to dairy cow feed additives. Detailed Implementation

[0017] The present invention will be further described in detail below with reference to embodiments, but the implementation of the present invention is not limited thereto. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in this technical field. Test methods in the following embodiments that do not specify specific experimental conditions are generally performed according to conventional experimental conditions or experimental conditions recommended by the manufacturer. Unless otherwise specified, the reagents and raw materials used in the present invention are commercially available.

[0018] Example 1 A feed additive for increasing the branched-chain fatty acid content in milk, comprising, by weight, the following ingredients: 3 parts leucine, 3 parts isoleucine, 3 parts valine, 2 parts *Symplocos edulis* extract, and 0.5 parts sodium subtilis lipopeptide.

[0019] The preparation method of the *Cynanchum paniculatum* extract is as follows: a 7wt% acetic acid aqueous solution is prepared in advance. The leaves of *Cynanchum paniculatum* are dried, crushed, and added to the acetic acid aqueous solution. The solid-liquid ratio of the leaves of *Cynanchum paniculatum* to the acetic acid aqueous solution is 1g:8mL. Then, the extract is ultrasonically extracted at 55℃ for 2h. The extract is collected, concentrated under reduced pressure, and then freeze-dried to obtain the *Cynanchum paniculatum* extract.

[0020] A method for preparing a feed additive that increases the content of branched-chain fatty acids in milk includes the following steps: leucine, isoleucine, valine, *Prickly ash* extract, and *Bacillus subtilis* lipopeptide sodium are mixed evenly according to the specified ratio to obtain the additive.

[0021] Example 2 A feed additive for increasing the branched-chain fatty acid content in milk, comprising, by weight, the following ingredients: 1 part leucine, 1 part isoleucine, 1 part valine, 1 part *Symplocos edulis* extract, and 0.3 parts sodium subtilis lipopeptide.

[0022] The preparation method of the *Cynanchum paniculatum* extract is as follows: Prepare a 5wt% acetic acid aqueous solution in advance, take the dried leaves of *Cynanchum paniculatum* and crush them, add them to the acetic acid aqueous solution, and the solid-liquid ratio of the leaves of *Cynanchum paniculatum* to the acetic acid aqueous solution is 1g:6mL; then, ultrasonically extract at 50℃ for 3h, collect the extract, concentrate under reduced pressure and freeze dry to obtain the *Cynanchum paniculatum* extract.

[0023] A method for preparing a feed additive that increases the content of branched-chain fatty acids in milk includes the following steps: leucine, isoleucine, valine, *Prickly ash* extract, and *Bacillus subtilis* lipopeptide sodium are mixed evenly according to the specified ratio to obtain the additive.

[0024] Example 3 A feed additive for increasing the branched-chain fatty acid content in milk, comprising, by weight, the following ingredients: 4 parts leucine, 4 parts isoleucine, 4 parts valine, 2.5 parts *Symplocos edulis* extract, and 0.6 parts sodium subtilis lipopeptide.

[0025] The preparation method of the *Cynanchum paniculatum* extract is as follows: Prepare an 8wt% acetic acid aqueous solution in advance, take the dried leaves of *Cynanchum paniculatum* and crush them, add them to the acetic acid aqueous solution, and the solid-liquid ratio of the leaves of *Cynanchum paniculatum* to the acetic acid aqueous solution is 1g:10mL; then, ultrasonically extract at 60℃ for 1h, collect the extract, concentrate under reduced pressure and freeze dry to obtain the *Cynanchum paniculatum* extract.

[0026] A method for preparing a feed additive that increases the content of branched-chain fatty acids in milk includes the following steps: leucine, isoleucine, valine, *Prickly ash* extract, and *Bacillus subtilis* lipopeptide sodium are mixed evenly according to the specified ratio to obtain the additive.

[0027] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that isoleucine and valine are omitted, and only leucine is added, with the amount of leucine adjusted to the sum of the three; otherwise, it remains the same as Example 1.

[0028] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that the extract of *Cynanchum paniculatum* tip is omitted; otherwise, they are the same as in Example 1.

[0029] Comparative Example 3 The difference between Comparative Example 3 and Example 1 is that sodium subtilisin was omitted; otherwise, they are the same as in Example 1.

[0030] Test case 1. Experimental animals and grouping Healthy Holstein cows with a weight of 620±30kg, parity of 2, lactation age of 140±16d, and similar milk production were selected. The cows were then divided into Example 1 group, Example 2 group, Example 3 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and control group, with 10 cows in each group.

[0031] 2. Animal husbandry methods Control group: Basal diet (by weight percentage, including: 12.85% flaked corn, 33.12% corn silage, 15.30% alfalfa, 5.26% dried distillers grains and their solubles, 4.08% whole cottonseed, 12.68% soybean meal, 1.00% extruded soybean, 11.85% wheat bran, 0.36% sodium bicarbonate, 3.50% premix, each kilogram of premix containing the following components: Vitamin A 200,000 IU, Vitamin D 65,000 IU, Vitamin E 2,100 IU, Iron 400 mg, Copper 560 mg, Zinc 2,100 mg, Manganese 590 mg, Selenium 15 mg, Iodine 40 mg, Cobalt 60 mg, Niacin 3,000 mg); Examples 1-3 and Comparative Examples 1-3: 0.8% (by weight of basal diet) of the corresponding feed additives for each group were added to the basal diet, mixed evenly, and then fed. The dairy cows in each group were fed three times a day (07:00, 14:00, and 18:00) with free access to feed and water, and milked three times a day (06:30, 13:30, and 20:30). The trial period lasted four weeks. Daily milk production was recorded. In the last week of the trial period, milk samples were collected daily, and the milk samples from each cow were mixed thoroughly in a mass ratio of morning:noon:evening of 4:3:3 to obtain a mixed milk sample.

[0032] 1. The milk composition in the mixed milk sample was determined using the FOSS milk composition analyzer (FT120), and the results are recorded in Table 1.

[0033] Table 1. Effects of different groups of feed additives on dairy cow production performance. As shown in Table 1, the milk yield, milk protein content, milk fat content, lactose content, total solids content, and non-fat solids content of the products obtained in Examples 1-3 of this invention are significantly higher than those in Comparative Examples 1-3. This indicates that the feed additive (containing three branched-chain amino acids, *Symplocos edulis* extract, and *Bacillus subtilis* lipopeptide sodium) can effectively improve milk yield and quality.

[0034] Example 1 showed the best results. Compared to Example 1, Comparative Example 1 showed a decrease in milk yield, indicating that the effect of using a single branched-chain amino acid is not as good as using three in combination. The three branched-chain amino acids have a synergistic effect in promoting mammary gland metabolism and fatty acid synthesis. Comparative Example 2 (without *Cynanchum paniculatum* extract) and Comparative Example 3 (without *Bacillus subtilis* lipopeptide sodium): milk yield, milk protein percentage, milk fat percentage, lactose percentage, total solids percentage, and non-fat solids percentage were all significantly lower than those of Example 1, indicating that *Cynanchum paniculatum* extract and *Bacillus subtilis* lipopeptide sodium have an important synergistic effect in increasing milk yield and can effectively improve the nutritional composition and quality of milk.

[0035] 2. Determination of BCFA content in milk Weigh a mixed milk sample, add a hexane-isopropanol mixture, vortex, centrifuge, and collect the upper organic phase (containing fat). Add 2% sodium hydroxide-methanol solution, heat in a 50℃ water bath for 30 minutes to convert triglycerides into fatty acid salts. Add 10% hydrochloric acid-methanol solution, heat in a 90℃ water bath for 2 hours to convert fatty acids into fatty acid methyl esters. After cooling, extract three times with water and hexane, combine the hexane layers, add anhydrous sodium sulfate, shake, and collect the supernatant for analysis. Determine the fatty acid content using gas chromatography. The results are recorded in Table 2.

[0036] Table 2. Effects of different feed additives on branched-chain fatty acids in milk. BCFAs have a unique branched-chain structure and are believed to improve the flavor and texture of dairy products. Furthermore, numerous studies have shown that consuming dairy-derived BCFAs (especially the odd-numbered carbon chain types iso and anteiso) is beneficial to human metabolic health.

[0037] As shown in Table 2, the performance of the products obtained in Examples 1-3 of this invention is superior to that of Comparative Examples 1-3. The contents of iso-C13:0, anteiso-C13:0, iso-C14:0, iso-C15:0, anteiso-C15:0, iso-C16:0, iso-C17:0, anteiso-C17:0, iso-C18:0, and total BCFA in milk are significantly increased. This indicates that the feed additive described in this invention can significantly and specifically increase the content of various BCFAs in milk. Compared to the control group, the fatty acid content of Comparative Examples 1-3 is increased, but the total BCFA in Comparative Example 1 is lower than that in Example 1, indicating that the effect of adding leucine alone is not as good as the combination of various ingredients, which is not conducive to the lactation function of dairy cows and the increase of BCFA. The total BCFA in Comparative Examples 2 and 3 is significantly lower than that in Example 1, indicating that the extract of *Cynanchum paniculatum* and sodium subtilis lipopeptide can optimize the overall absorption and utilization environment and have a positive promoting effect on increasing the content of various branched-chain fatty acids in milk. The above test results show that the combination of three amino acids (isoleucine, valine, and leucine), *Symplocos edulis* extract, and *Bacillus subtilis* lipopeptide sodium in the feed additive of this invention can increase the content of branched-chain fatty acids in milk, improve milk quality, and enhance the milk production performance of dairy cows.

[0038] The above embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention shall fall within the scope of protection claimed by the present invention.

Claims

1. A feed additive for increasing the branched-chain fatty acid content in milk, characterized in that, By weight, it includes the following ingredients: 1-4 parts leucine, 1-4 parts isoleucine, 1-4 parts valine, 1-2.5 parts *Symplocos edulis* extract, and 0.3-0.6 parts sodium subtilis lipopeptide. The preparation method of the *Cynanchum paniculatum* extract is as follows: the leaves of *Cynanchum paniculatum* are dried, pulverized, added to an aqueous acetic acid solution, ultrasonically extracted, the extract is collected, concentrated under reduced pressure, and then freeze-dried to obtain the *Cynanchum paniculatum* extract.

2. The feed additive for increasing the branched-chain fatty acid content in milk according to claim 1, characterized in that, By weight, it includes the following ingredients: 3 parts leucine, 3 parts isoleucine, 3 parts valine, 1.5 parts *Symplocos edulis* extract, and 0.5 parts sodium subtilis lipopeptide.

3. The feed additive for increasing the branched-chain fatty acid content in milk according to claim 1, characterized in that, The solid-liquid ratio of the leaves of the *Cynanchum paniculatum* to the aqueous acetic acid solution is 1g:6-10mL.

4. The feed additive for increasing the branched-chain fatty acid content in milk according to claim 1, characterized in that, The ultrasonic extraction temperature is 50-60℃; the time is 1-3 hours.

5. The feed additive for increasing the branched-chain fatty acid content in milk according to claim 1, characterized in that, The acetic acid aqueous solution has an acetic acid mass concentration of 5-8%.

6. A method for preparing a feed additive to increase the branched-chain fatty acid content in milk according to any one of claims 1-5, characterized in that, The process includes the following steps: According to the specified ratio, leucine, isoleucine, valine, *Prickly ash* extract, and *Subtilis lipopeptide sodium* are stirred and mixed evenly to obtain the final product.

Images