Biological fermentation type premix for beef cattle as well as preparation method and application thereof

Abstract

The invention relates to the technical field of livestock breeding, in particular to a biological fermentation type premix for beef cattle as well as a preparation method and application thereof. The invention provides a biological fermentation type premix. The biological fermentation type premix comprises a basic component, a fermentation component, an essential oil component and a trace element premix. The basic components comprise conventional feed raw materials such as corn flour, soybean meal and wheat bran, and unconventional feed raw materials such as grape seed extract, wheat germs and distiller's grains; the fermentation components comprise beneficial bacteria such as bacillus subtilis, lactic acid bacteria, saccharomycetes and streptomyces fusiformis and fermentation products of the beneficial bacteria; the essential oil component comprises parsley essential oil and eucommia ulmoides essential oil; and the trace element premix contains a plurality of mineral substances. The biological fermentation type premix provided by the invention is balanced in nutrition, can meet the nutritional requirements of beef cattle in different growth stages, and is beneficial to promoting the rapid growth of the beef cattle and improving the meat quality; the aims of relieving rumen acidosis of the beef cattle and improving the immunity of the beef cattle can also be fulfilled.

Description

Technical Field

[0001] The invention relates to the technical field of animal husbandry, and in particular to a biological fermentation premix for beef cattle, and a preparation method and application thereof. Background Art

[0002] In recent years, with the rapid development of cattle breeding, large-scale beef cattle breeding has become more and more large-scale, scientific and economical. The amount of beef cattle raised accounts for a considerable proportion, especially in provinces and regions with more developed agriculture, most breeders adopt centralized feeding. Various beef cattle feeds have also developed rapidly. At present, there are many varieties of cattle feeds on the market with different functions. Among them, cattle have different nutritional requirements in different growth and physiological periods, and the feed they feed cannot be the same, and needs to be specially formulated.

[0003] Beef cattle rely on humans for their food throughout their lives, so feed composition and feeding methods are particularly important. At the same time, they should be adjusted in time according to their different growth stages in order to breed better quality and healthier beef cattle. At present, the feed formula for beef cattle breeding includes: (1) Configuration of roughage: Generally, roughage is mainly composed of raw materials such as silage corn stalks, brewer's grains and white wine grains, and the requirements for fine-grained proportion adjustment are not high. At the same time, it can meet the goal of feeding throughout the year and break the time limit. (2) Formulate the feed according to the weight of the beef cattle: When the weight of the beef cattle is 400 kg, the proportion of corn in the feed is 65%, cottonseed cake is 15%, bran is 11.5%, bean cake is 5%, bone meal, salt and baking soda are 1%, and flavoring agent is 0.5%; when the beef cattle belong to the young herd and the actual weight is more than 250 kg, it is necessary to make reasonable adjustments to the corresponding feed formula: corn is 60%, cottonseed cake is 20%, bean cake is 10%, bran is 5.5%, bone meal is 1.5%, and salt, baking soda and flavoring agent are 1% each.

[0004] With the rapid development of animal husbandry, the quality and efficiency requirements of beef cattle feed are increasing. Although traditional premixes meet the nutritional needs of beef cattle to a certain extent, they still need to be improved in promoting rapid growth of beef cattle, alleviating rumen acidosis, improving immunity and improving meat quality. Therefore, the development of a new type of bio-fermentation premix is ​​of great significance to improving the comprehensive performance and use effect of beef cattle compound premix feed. Summary of the invention

[0005] The object of the present invention is to provide a bio-fermented premix for beef cattle, its preparation method and application, so as to solve the problems existing in the above-mentioned prior art. The present invention provides a bio-fermented premix, the composition of which includes a basic component, a fermentation component, an essential oil component and a trace element premix. This bio-fermented premix can effectively promote the rapid growth of beef cattle, relieve rumen acidosis, improve immunity and improve meat quality.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The present invention provides a bio-fermented premix for beef cattle, including the following components in parts by mass:

[0008] 20-40 parts of corn flour, 10-20 parts of soybean meal, 5-10 parts of wheat bran, 8-20 parts of grape seed extract, 3-10 parts of wheat germ, 2-10 parts of distillers' grains, 3-4 parts of parsley essential oil, 2-3 parts of eucommia essential oil, 5-10 parts of fermentation product and 1-5 parts of trace element premix;

[0009] The preparation method of the fermentation product includes the step of fermenting and culturing fermentation strains to obtain the fermentation product; the fermentation strains include Bacillus subtilis, lactic acid bacteria, yeast and Streptomyces fusantomycin.

[0010] Preferably, the bio-fermented premix includes the following components in parts by mass:

[0011] 30 parts of corn flour, 15 parts of soybean meal, 8 parts of wheat bran, 15 parts of grape seed extract, 7 parts of wheat germ, 6 parts of distillers' grains, 3 parts of parsley essential oil, 3 parts of eucommia essential oil, 8 parts of fermentation product and 3 parts of trace element premix.

[0012] Preferably, the temperature of the fermentation culture is 28-42 °C, the time is 64-72 h, the initial pH value is 5.7-6.8, the water content is 37%-40%, and the inoculation amount is 1%-2%.

[0013] Preferably, the trace element premix includes the following components:

[0014] 50-200 mg / kg of copper, 100-2000 mg / kg of iron, 500-3000 mg / kg of zinc and 50-300 mg / kg of selenium.

[0015] The present invention provides a preparation method of the above bio-fermented premix, including the following steps:

[0016] Mix the corn flour, the soybean meal, the wheat bran, the grape seed extract, the wheat germ and the distillers' grains evenly to obtain a basic feed;

[0017] Mix the parsley essential oil and the eucommia ulmoides essential oil evenly to obtain an essential oil combination;

[0018] Ferment the fermentation strain to obtain the fermentation product; the fermentation strain includes Bacillus subtilis, lactic acid bacteria, yeast, and Streptomyces fusiformis;

[0019] Mix the basal feed, the essential oil combination, the fermentation product, and the trace element premix to obtain the biological fermentation type premix.

[0020] Preferably, the temperature of the fermentation culture is 28 - 42 °C, the time is 64 - 72 h, the initial pH value is 5.7 - 6.8, the water content is 37% - 40%, and the inoculation amount is 1% - 2%.

[0021] More preferably, the trace element premix includes the following components:

[0022] Copper 50 - 200 mg / kg, iron 100 - 2000 mg / kg, zinc 500 - 3000 mg / kg, and 50 - 300 mg / kg.

[0023] The present invention provides the application of the above biological fermentation type premix in the preparation of a feed additive or feed for beef cattle.

[0024] The present invention provides a feed additive for beef cattle, and the feed additive includes the above biological fermentation type premix.

[0025] The present invention provides a feed for beef cattle, and the feed includes the above biological fermentation type premix.

[0026] The present invention provides the application of the above biological fermentation type premix, the above feed additive, or the above feed in any one or several of the following:

[0027] (1) Application in improving the growth of beef cattle;

[0028] (2) Application in alleviating rumen acidosis in beef cattle;

[0029] (3) Application in improving the immunity of beef cattle;

[0030] (4) Application in improving the feed nutrient digestibility of beef cattle.

[0031] As an additional aspect, the present invention provides the application of the above biological fermentation type premix in the preparation of a drug for alleviating rumen acidosis in beef cattle.

[0032] As an additional aspect, the present invention provides a drug for alleviating rumen acidosis in beef cattle, and the drug includes the above biological fermentation type premix.

[0033] The present invention discloses the following technical effects:

[0034] The present invention provides a bio-fermented premix for beef cattle, which comprises a basic component, a fermentation component, an essential oil component and a trace element premix. The basic component includes conventional feed raw materials such as corn flour, soybean meal, wheat bran, etc. and unconventional feed raw materials such as grape seed extract, wheat germ and distillers grains; the fermentation component includes beneficial bacteria such as Bacillus subtilis, Lactobacillus, Saccharomyces cerevisiae and Streptomyces fusiformis and their fermentation products; the essential oil component includes parsley essential oil and eucommia ulmoides essential oil; and the trace element premix contains various minerals to meet the comprehensive nutritional needs of beef cattle. Thus, the bio-fermented premix provided by the present invention has a balanced nutrition, can meet the nutritional needs of beef cattle at different growth stages, helps to promote the rapid growth of beef cattle and improve the meat quality; it can also relieve rumen acidosis and improve the immunity.

[0035] The present invention also provides a preparation method of the bio-fermented premix. By means of the bio-fermentation technology, the present invention improves the content of strain metabolites in the premix, especially the content and activity of prebiotics such as mannan, organic acids, oligopeptides, etc. and beneficial bacteria, which helps to promote the intestinal health of beef cattle and improve the immunity. And the preparation method is simple and easy to operate, with low cost, and is suitable for large-scale production and application. Detailed Embodiments

[0036] Now, various exemplary embodiments of the present invention will be described in detail. This detailed description should not be construed as a limitation on the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention.

[0037] It should be understood that the terms described in the present invention are only for describing particular embodiments and are not used to limit the present invention. In addition, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded within the range.

[0038] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes the preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0039] Without departing from the scope or spirit of the present invention, various modifications and variations can be made to the specific embodiments of the description of the present invention, which will be apparent to those skilled in the art. Other embodiments obtained from the description of the present invention will be apparent to those skilled in the art. The description and examples of the present invention are merely exemplary.

[0040] Regarding "comprising", "including", "having", "containing", etc. used herein, they are all open-ended terms, meaning including but not limited to.

[0041] Unless otherwise specified, the raw materials used in the present invention are all routinely purchased by those skilled in the art, and the methods used are all methods well-known to those skilled in the art.

[0042] Example 1 Basic Components and Preparation Method of Bio-Fermented Premix

[0043] A bio-fermented premix is composed of the following components in parts by mass:

[0044] Basic components: 20 - 40 parts of corn flour, 10 - 20 parts of soybean meal, 5 - 10 parts of wheat bran, 8 - 20 parts of grape seed extract, 3 - 10 parts of wheat germ, and 2 - 10 parts of distillers' grains; among them, the distillers' grains are provided by Hebei Huaren Biotechnology Co., Ltd.

[0045] Plant essential oil components: 3 - 4 parts of parsley essential oil and 2 - 3 parts of eucommia ulmoides essential oil;

[0046] Fermentation components: 5 - 10 parts of fermentation products; the parts by mass of the strains used in the preparation of the fermentation components are as follows: 0.2 - 0.6 parts of Bacillus subtilis (bacterial activity is 10 7 CFU / g or bacterial activity is 10 7 CFU / g), 0.1 - 0.2 parts of lactic acid bacteria (bacterial activity is 10 7 CFU / g or bacterial activity is 10 7 CFU / g), 0.1 - 0.3 parts of yeast (bacterial activity is 10 7 CFU / g or bacterial activity is 10 7 CFU / g) and 0.2 - 0.5 parts of Streptomyces spondylomycini (purchased from Youlike (Shanghai) Life Sciences Co., Ltd., product number YLK-J3264h, bacterial activity is 10 7 CFU / g or bacterial activity is 10 7 CFU / g);

[0047] Trace element components: 1 - 5 parts of trace element premix, and the trace element premix contains 50 - 200 mg / kg of copper, 100 - 2000 mg / kg of iron, 500 - 3000 mg / kg of zinc, and 50 - 300 mg / kg of selenium.

[0048] The preparation method of this biological fermentation type premix is as follows:

[0049] Mix the basic components and the plant essential oil components respectively to obtain a basic feed and an essential oil combination;

[0050] Inoculate the fermentation strain into a self-developed culture medium (the components consist of a carbon source, a nitrogen source, inorganic elements, trace elements, growth factors, a complex enzyme preparation and water. Among them, the carbon source: corn flour / wheat bran 5-20 wt.%, molasses 2-10 wt.%, glucose 1-5 wt.%, straw 10-30 wt.%; the nitrogen source: urea 0.5-2 wt.%, ammonium nitrate / ammonium sulfate 0.5-1.5 wt.%, soybean meal 5-15 wt.%, fish meal / yeast extract powder 1-5 wt.%; inorganic elements: potassium dihydrogen phosphate 0.1-0.5 wt.%, magnesium sulfate 0.05-0.2 wt.%, sodium chloride 0.1-0.3 wt.%, calcium salt 0.5-2 wt.%; trace elements: zinc 0.01-0.1 wt.%, iron 0.01-0.1 wt.%, manganese 0.01-0.1 wt.%; growth factors: yeast extract / yeast powder 0.5-2 wt.%; complex enzyme preparation: cellulase (1000 U / mL) 0.1-0.5 wt.%, amylase (1000 U / mL) 0.1-0.5 wt.%; the balance is water) for fermentation culture to obtain a fermentation product, and the particle size of the fermentation product is 0.8 mm (ensuring uniform particle size of the material and avoiding too large or too small particles from affecting the fermentation efficiency of yeast. The crushed material should be able to pass through a sieve hole of a certain size (such as 20 mesh or 40 mesh) to ensure that its particle size meets the requirements); during the fermentation culture process, the condition parameters that need to be controlled are as follows:

[0051] (1) Control the moisture content of the fermentation environment between 37% and 40%. The moisture content has an important impact on the activity of the fermentation strain (especially yeast). Within this range of moisture, the growth and metabolism of the fermentation strain (especially yeast) can reach the best state. During the fermentation process, the moisture content should be regularly detected and adjusted as needed.

[0052] (2) Control the initial pH value between 5.7 and 6.8. The initial pH value has an important impact on the growth of the fermentation strain (especially yeast) and the accumulation of the fermentation product. Within this range, the fermentation strain (especially yeast) can adapt and grow better. However, different types of fermentation strains (especially yeast) may have different adaptabilities to the pH value, so it should be adjusted according to the specific type of fermentation strain (especially yeast). In this embodiment, a buffer (such as potassium dihydrogen phosphate and dipotassium hydrogen phosphate) can be used to stabilize the pH value during the fermentation process. According to the change of the pH value during the fermentation process, acidic or alkaline substances are added in a timely manner for adjustment.

[0053] (3) The temperature is controlled between 28 - 42 °C, and more preferably between 32 - 38 °C. Temperature is one of the key factors affecting the activity of fermentation strains (especially yeast). Within this range, the fermentation strains (especially yeast) can maintain a relatively high activity. However, if the temperature is too high (exceeding 42 °C), it will cause the inactivation of fermentation strains (especially yeast), affecting the fermentation effect. Therefore, the temperature during the fermentation process should be strictly controlled. When the temperature is within the range of 32 - 38 °C, the metabolic activities of fermentation strains (especially yeast) are the most vigorous, which is conducive to the accumulation of metabolites. A heating or cooling system should be used to maintain the temperature in the fermentation tank within the optimal range. At the same time, the temperature should be regularly detected and adjusted as needed.

[0054] (4) The anaerobic fermentation cycle is controlled between 64 - 72 h. Anaerobic fermentation is the main stage for fermentation strains (especially yeast) to produce metabolites. Within this time range, the fermentation strains (especially yeast) can fully carry out anaerobic respiration and accumulate metabolites. However, the length of the fermentation cycle is also affected by factors such as the type of fermentation strain, fermentation substrate, and fermentation conditions. Therefore, it should be adjusted according to specific circumstances.

[0055] (5) During the fermentation process, key indicators (such as pH value, oxygen concentration, metabolite concentration, etc.) should be regularly detected to determine whether the fermentation is completed. When these indicators reach the preset values, it can be considered that the fermentation has ended.

[0056] (6) Other parameters

[0057] (6.1) Nutritional components: The fermentation medium should contain the nutritional components required for yeast growth, such as carbon sources (glucose, brown sugar, etc.), nitrogen sources (peptone, yeast powder, etc.), inorganic salts (potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate crystals, etc.), and trace elements. According to the type of fermentation strain (especially yeast) and the requirements of fermentation products, the formula of the medium can be adjusted to obtain the best fermentation effect.

[0058] (6.2) An appropriate inoculation amount can ensure that the yeast in the fermentation strain grows rapidly and occupies a dominant position in the initial stage of fermentation. The inoculation amount is usually controlled between 1% - 2% of the substrate (i.e., 0.2 - 0.6 parts of Bacillus subtilis, 0.1 - 0.2 parts of lactic acid bacteria, 0.1 - 0.3 parts of yeast, and 0.2 - 0.5 parts of Streptomyces fusantomycin). However, the specific inoculation amount is also affected by factors such as yeast type, fermentation substrate, and fermentation conditions.

[0059] (6.3) Stirring and aeration: In the initial stage of fermentation, appropriate stirring can promote the growth and reproduction of fermentation strains (especially yeast) and the uniform distribution of nutrients. However, in the anaerobic fermentation stage, the aeration volume should be reduced to avoid adverse effects on fermentation strains (especially yeast). The stirring speed and time should be adjusted according to specific circumstances to ensure the growth and fermentation effect of fermentation strains (especially yeast). In this example, stirring is carried out once every 4 - 12 h, for 5 - 15 min each time, and the rotation speed is 10 - 30 rpm; the condition parameters of aeration are: aerobic fermentation 0.5 - 2 vvm, microaerobic / facultative anaerobic fermentation 0.05 - 0.2 vvm, air composition: 21% oxygen + 79% nitrogen, and some fungal fermentations require carbon dioxide induction for metabolism, with a need for 2 - 10% carbon dioxide; in the aeration mode, surface aeration is 0.05 - 0.1 vvm, and the deep aeration pressure requirement is 0.1 - 0.3 M pa.

[0060] (6.4) Fermentation substrate: The fermentation substrate should not exceed one - quarter of the total filling volume to ensure sufficient growth space and nutrients for yeast. At the same time, the type and concentration of the substrate will also affect the growth and fermentation effect of yeast. Therefore, appropriate substrates should be selected and optimized according to specific circumstances.

[0061] (6.5) Sterilization and aseptic operation: During the fermentation process, the aseptic state of all equipment and culture media should be ensured to avoid contamination by miscellaneous bacteria. Methods such as high - pressure steam sterilization can be used to sterilize equipment and culture media. At the same time, attention should be paid to aseptic operation specifications during the operation process to prevent contamination.

[0062] (6.6) Monitoring and control during fermentation: During the fermentation process, key indicators (such as temperature, pH value, oxygen concentration, metabolite concentration, etc.) should be monitored regularly to understand the fermentation process and effect. And the fermentation parameters should be adjusted in a timely manner according to the monitoring results to ensure the best fermentation effect. At the same time, the data during the fermentation process should be recorded and analyzed to summarize experience and optimize the fermentation process.

[0063] Mix the basic feed, essential oil combination, fermentation components and trace element components to obtain a bio - fermented premix.

[0064] Example 2

[0065] A bio - fermented premix is composed of the following components in parts by mass:

[0066] Basic components: 30 parts of corn flour, 15 parts of soybean meal, 8 parts of wheat bran, 15 parts of grape seed extract, 7 parts of wheat germ and 6 parts of distiller's grains;

[0067] Plant essential oil components: 3 parts of parsley essential oil and 3 parts of eucommia essential oil;

[0068] Fermentation components: 8 parts of fermentation products; the mass parts of the strains used in the preparation of the fermentation components are as follows: 0.4 parts of Bacillus subtilis, 0.1 part of lactic acid bacteria, 0.2 part of yeast, and 0.4 part of Streptomyces spondylomycini;

[0069] Trace element components: 3 parts of trace element premix, and the trace element premix contains 200 mg / kg of copper, 100 mg / kg of iron, 3000 mg / kg of zinc, and 200 mg / kg of selenium.

[0070] The preparation method of the biological fermentation type premix is as follows:

[0071] Mix the basic components and the plant essential oil components respectively to obtain the basic feed and the essential oil combination;

[0072] Inoculate the fermentation strain into a self-developed culture medium (the components are composed of carbon source, nitrogen source, inorganic elements, trace elements, growth factors, complex enzyme preparations and water. Among them, carbon source: 15 wt.% of corn flour, 7 wt.% of molasses, 3 wt.% of glucose, 20 wt.% of straw; nitrogen source: 1 wt.% of urea, 1 wt.% of ammonium nitrate / ammonium sulfate, 10 wt.% of soybean meal, 3 wt.% of yeast extract powder; inorganic elements: 0.3 wt.% of potassium dihydrogen phosphate, 0.1 wt.% of magnesium sulfate, 0.2 wt.% of sodium chloride, 1 wt.% of calcium salt; trace elements: 0.03 wt.% of zinc, 0.03 wt.% of iron, 0.03 wt.% of manganese; growth factors: 1 wt.% of yeast extract; complex enzyme preparations: 0.2 wt.% of cellulase (1000 U / mL), 0.2 wt.% of amylase (1000 U / mL); the balance is water) for fermentation culture to obtain the fermentation product, and the particle size of the fermentation product is 0.8 mm; during the fermentation culture process, the following condition parameters need to be controlled:

[0073] (1) The moisture content of the fermentation environment is controlled between 38% and 39%.

[0074] (2) The initial pH value is controlled between 6.0 and 6.5.

[0075] (3) The temperature is controlled between 32 and 38 °C.

[0076] (4) The anaerobic fermentation period is controlled between 64 and 72 h.

[0077] (5) During the fermentation process, key indicators (such as pH value, oxygen concentration, metabolite concentration, etc.) should be regularly detected to judge whether the fermentation is over. When these indicators reach the preset values, it can be considered that the fermentation is over.

[0078] (6) Other parameters

[0079] (6.1) The inoculum size is 1% (i.e., 0.4 parts of Bacillus subtilis, 0.1 part of lactic acid bacteria, 0.2 parts of yeast, and 0.4 parts of Streptomyces fusantomycin).

[0080] (6.2) Stirring and aeration:

[0081] (6.3) Stirring and aeration: Stir once every 8 h, each time for 10 min, and the rotation speed is 20 rpm; the condition parameters of aeration are as follows: for aerobic fermentation, 1 vvm; for microaerobic / facultative anaerobic fermentation, 0.1 vvm; air composition: 21% oxygen + 79% nitrogen. For the fermentation of some fungi, carbon dioxide induction metabolism is required, and 8% carbon dioxide is needed; in the aeration mode, surface aeration is 0.05 - 0.1 vvm, and the deep aeration pressure requirement is 0.2 MPa.

[0082] (6.4) Fermentation substrate: The fermentation substrate should not exceed one - quarter of the total loading volume.

[0083] (6.5) Sterilization and aseptic operation: During the fermentation process, the aseptic state of all equipment and culture media should be ensured to avoid contamination by miscellaneous bacteria. Methods such as high - pressure steam sterilization can be used to sterilize the equipment and culture media. At the same time, aseptic operation specifications should be noted during the operation to prevent contamination.

[0084] (6.6) Monitoring and control during the fermentation process: During the fermentation process, key indicators (such as temperature, pH value, oxygen concentration, metabolite concentration, etc.) should be regularly monitored to understand the fermentation process and effect. And the fermentation parameters should be adjusted in a timely manner according to the monitoring results to ensure the best fermentation effect.

[0085] Mix the basal feed, essential oil combination, fermentation components, and trace element components to obtain a bio - fermented premix.

[0086] Example 3

[0087] A bio - fermented premix is composed of the following components in parts by mass:

[0088] Basal components: 40 parts of corn flour, 10 parts of soybean meal, 10 parts of wheat bran, 8 parts of grape seed extract, 10 parts of wheat germ, and 2 parts of distillers' grains;

[0089] Plant essential oil components: 3 parts of parsley essential oil and 2 parts of eucommia essential oil;

[0090] Fermentation components: 10 parts of fermentation products; the parts by mass of the strains used in the preparation of the fermentation components are as follows: 0.6 part of Bacillus subtilis, 0.2 part of lactic acid bacteria, 0.1 part of yeast, and 0.5 part of Streptomyces fusantomycin;

[0091] Trace element components: 5 parts of trace element premix, which contains 50 mg / kg of copper, 2000 mg / kg of iron, 500 mg / kg of zinc and 200 mg / kg of selenium.

[0092] The preparation method of this bio-fermented premix is the same as that of Example 2.

[0093] Example 4

[0094] A bio-fermented premix, which is composed of the following components in parts by mass:

[0095] Base components: 20 parts of corn flour, 20 parts of soybean meal, 5 parts of wheat bran, 20 parts of grape seed extract, 3 parts of wheat germ and 10 parts of distillers grains;

[0096] Plant essential oil components: 4 parts of parsley essential oil and 3 parts of eucommia essential oil;

[0097] Fermentation components: 5 parts of fermentation products; The parts by mass of the strains used in the preparation of the fermentation components are as follows: 0.2 parts of Bacillus subtilis, 0.1 part of lactic acid bacteria, 0.3 part of yeast and 0.2 part of Streptomyces fusiformis.

[0098] Trace element components: 1 part of trace element premix, which contains 200 mg / kg of copper, 100 mg / kg of iron, 3000 mg / kg of zinc and 200 mg / kg of selenium.

[0099] The preparation method of this bio-fermented premix is the same as that of Example 2.

[0100] Control 1

[0101] A bio-fermented premix, which is composed of the following components in parts by mass:

[0102] Base components: 30 parts of corn flour, 15 parts of soybean meal, 8 parts of wheat bran, 15 parts of grape seed extract, 7 parts of wheat germ and 6 parts of distillers grains;

[0103] Fermentation components: 10 parts of fermentation products; The parts by mass of the strains used in the preparation of the fermentation components are as follows: 0.4 part of Bacillus subtilis, 0.1 part of lactic acid bacteria and 0.2 part of yeast;

[0104] Trace element premix: 3 parts of trace element premix, which contains 200 mg / kg of copper, 100 mg / kg of iron, 3000 mg / kg of zinc and 200 mg / kg of selenium.

[0105] The preparation method of this bio-fermented premix is the same as that of Example 2.

[0106] Control 2

[0107] Base components: 30 parts of corn flour, 15 parts of soybean meal, 8 parts of wheat bran, 15 parts of grape seed extract, 7 parts of wheat germ, and 6 parts of distillers' grains;

[0108] Plant essential oil components: 3 parts of parsley essential oil and 3 parts of eucommia essential oil;

[0109] Fermentation components: 10 parts of fermentation product; The mass parts of the strains used in the preparation of the fermentation components are as follows: 0.4 parts of Bacillus subtilis, 0.1 part of lactic acid bacteria, 0.2 part of yeast, and 0.4 part of Streptomyces spondylomyceticus;

[0110] Trace element premix: 3 parts of trace element premix, and this trace element premix contains 200 mg / kg of copper, 100 mg / kg of iron, 3000 mg / kg of zinc, and 200 mg / kg of selenium.

[0111] The preparation method of this bio-fermented premix is the same as that of Example 2.

[0112] Comparative Example 3

[0113] Base components: 30 parts of corn flour, 15 parts of soybean meal, 8 parts of wheat bran, 15 parts of grape seed extract, 7 parts of wheat germ, and 6 parts of distillers' grains;

[0114] Plant essential oil components: 3 parts of parsley essential oil and 3 parts of eucommia essential oil;

[0115] Fermentation components: 10 parts of fermentation product; The mass parts of the strains used in the preparation of the fermentation components are as follows: 0.4 parts of Bacillus subtilis, 0.1 part of lactic acid bacteria, and 0.2 part of yeast;

[0116] Trace element premix: 3 parts of trace element premix, and this trace element premix contains 200 mg / kg of copper, 100 mg / kg of iron, 3000 mg / kg of zinc, and 200 mg / kg of selenium.

[0117] The preparation method of this bio-fermented premix is the same as that of Example 2.

[0118] Effect Example 1

[0119] 1. Methods of test materials

[0120] 1.1. Bio-fermented premix: The bio-fermented premixes prepared in Examples 2-4 and Comparative Examples 1-3.

[0121] 1.2. Test grouping

[0122] The experiment was conducted in 2023. Seventy beef cattle with similar body weights and disease-free health were selected and evenly divided into 7 treatment groups according to a single-factor experimental design, with 10 beef cattle in each group. The beef cattle in each group were fed the bio-fermented premixes prepared in Examples 2-4 and Comparative Examples 1-3 for 60 days with free access to food; at the same time, the treatment of feeding the basal diet (the total mixed ration formula in Table 1 without the bio-fermented premix) was used as the control group. The total mixed ration of the beef cattle in this experiment was formulated with reference to the "Feeding Standard for Beef Cattle" (NY / T 815-2004) in China. The concentrate-to-forage ratio of the ration was 5:5, and its raw material composition and nutrient content are shown in Table 1.

[0123] Table 1 Total mixed ration formula for experimental beef cattle

[0124]

[0125]

[0126] Note: The nutritional level is a theoretical calculated value.

[0127] 1.3 Beef cattle breeding management

[0128] Before the start of the experiment, the breeding site and breeding equipment were disinfected. The experimental beef cattle were raised in the same breeding shed and tied up in separate pens. They were fed twice a day at 7:30 in the morning and 17:30 in the afternoon, and the beef cattle had free access to food and water. Immunization and deworming were carried out according to the daily norms, and the cattle shed was cleaned every day.

[0129] 1.4 Growth performance detection method

[0130] On the first day and the last day of the experiment, before the morning feeding of the beef cattle, the body weights of the beef cattle were measured on an empty stomach, and the feed intakes of the beef cattle in each group during the experimental period were accurately counted, which were used for statistical analysis of the average daily gain and feed-to-gain ratio data. Average daily gain = (final weight - initial weight) / experimental time; average daily feed intake = total feed intake of each group / experimental time × number of experimental beef cattle; feed-to-gain ratio = average daily feed intake / average daily gain.

[0131] 2. Test results

[0132] The data on the effect of the bio-fermented premix on the growth performance of beef cattle are shown in Table 2. From the data recorded in Table 2, it can be seen that the effects of Examples 1-3 are the best. The effect of Comparative Example 1 (omitting the essential oil component and one fermentation strain) is lower than that of Examples 1-3. In particular, it can be seen that adding the essential oil component and the new fermentation strain increases the average daily feed intake of beef cattle and reduces the feed-to-weight ratio. In Comparative Example 2 and Comparative Example 3, the essential oil component or the new fermentation strain was omitted respectively, which may lead to poor taste of the obtained bio-fermented premix, resulting in a decrease in the average daily feed intake, and ultimately leading to a decrease in the weight gain and feed-to-weight ratio of beef cattle. After comparison, it can be seen that the bio-fermented premix prepared in Example 2 has the best effect on promoting the growth of beef cattle. Therefore, the bio-fermented premix prepared in Example 2 was used for subsequent experiments.

[0133] Table 2 Effect of Bio-Fermented Premix on Growth Performance of Beef Cattle

[0134]

[0135] Effect Example 2

[0136] 1. Test Materials and Methods

[0137] 1.1 Bio-Fermented Premix

[0138] The bio-fermented premix used in this experiment was the bio-fermented premix prepared in Example 2.

[0139] 1.2 Test Grouping

[0140] In the experiment conducted in 2024, 60 beef cattle with similar body weights and disease-free health were selected and evenly divided into 4 treatment groups according to the single-factor experimental design, with 15 beef cattle in each group. The beef cattle in each group were fed a total mixed ration formula containing 0 g / (head·d) (control group), 20 g / (head·d) (Group 1), 40 g / (head·d) (Group 2), and 80 g / (head·d) (Group 3) of the bio-fermented premix for 60 days. The total mixed ration for the beef cattle in this experiment was formulated with reference to the "Feeding Standard for Beef Cattle" (NY / T 815-2004) in China. The concentrate-to-roughage ratio of the ration was 5:5, and its raw material composition and nutrient content are shown in Table 1.

[0141] 1.3 Beef Cattle Breeding Management

[0142] Same as Effect Example 1.

[0143] 1.4 Index Detection Method

[0144] Growth Performance:

[0145] The detection method for growth performance was the same as that in Effect Example 1.

[0146] Rumen fermentation parameters:

[0147] On the last day of the experiment, rumen fluid of beef cattle was collected through the oral cavity to detect rumen fermentation parameters. The pH value was measured using a pH meter; the NH3-N content was determined according to the reference "Effects of Different Addition Amounts of Lactic Acid Bacteria and Yeast and Fermentation Days on the Fermentation Quality and Nutritional Value of Sugarcane Tail Silage" (Peng Lijuan, Li Mengwei, Yang Chengjian, et al. Effects of Different Addition Amounts of Lactic Acid Bacteria and Yeast and Fermentation Days on the Fermentation Quality and Nutritional Value of Sugarcane Tail Silage [J]. Animal Husbandry & Veterinary Medicine, 2022, 54(3): 21-27.);

[0148] The MCP content was determined according to the reference "Potential benefits of a blend of essential oils on metabolism, digestibility, organ development and gene expression of dairy calves" (CAMPOLINA J P, COELHO S G, BELL I A L, et al. Potential benefits of a blend of essential oils on metabolism, digestibility, organ development and gene expression of dairy calves [J]. Scientific Reports, 2023, 13(1): 3378.); the VFA content was determined according to the reference "Silage review: interpretation of chemical, microbial, and organoleptic components of silage" (KUNG L, SHAVER R D, GRANT R J, et al. Silage review: interpretation of chemical, microbial, and organoleptic components of silages [J]. Journal of Dairy Science, 2018, 101(5): 4020-4033.).

[0149] Immune indexes:

[0150] Before the morning feeding of beef cattle on the last day of the experiment, blood samples were collected from the tail vein of beef cattle. After standing for 3 - 4 hours, the samples were centrifuged at 4000 r / min for 6 minutes, and the upper serum was taken for detecting immunoglobulin and cytokine indexes, mainly including IgA, IgG, IgM, IL-2, IL-6, IL-10, etc. The detection kits were provided by Nanjing Jiancheng Bioengineering Institute.

[0151] Nutrient digestibility:

[0152] On the 57th - 59th day of the experiment, a digestion experiment of beef cattle was carried out using the acid-insoluble ash (AIA) method. Feed samples and fecal samples of beef cattle were continuously collected for 3 days. Each feed sample was collected 200 g each time. Five beef cattle were selected from each group to collect their feces, 100 g each time. The feces corresponding to the 3 days were mixed evenly, 10 mL of dilute sulfuric acid was added for nitrogen fixation, and they were stored in a refrigerator at 4℃ for later use. Referring to the experimental method in the reference "Effects of Saccharomyces cerevisiae and Lactobacillus on the Digestive Metabolism and Growth Performance of Tan Sheep Fed with Corn Stover" (Ma Bucang. Effects of Saccharomyces cerevisiae and Lactobacillus on the Digestive Metabolism and Growth Performance of Tan Sheep Fed with Corn Stover [D]. Master's Thesis. Yinchuan: Ningxia University, 2023.), the digestibility of nutrients such as dry matter, crude protein, crude fat, neutral detergent fiber, acid detergent fiber, calcium, and phosphorus in feed and feces was detected. The calculation formula is as follows: Apparent digestibility of a certain nutrient (%) = 100 - 100×(AIA content in the diet / AIA content in the fecal sample)×(content of this nutrient in the fecal sample / content of this nutrient in the diet).

[0153] 2. Experimental results

[0154] 2.1 Effects of bio-fermented premix on the growth performance of beef cattle

[0155] From the data in Table 3, it can be seen that the average daily gain of beef cattle in Group 2 and Group 3 was significantly increased by 20.93% and 25.58% compared with the control group (P < 0.05); the feed-to-gain ratio was decreased by 10.45% and 11.85% compared with the control group (P < 0.05).

[0156] Table 3 Effects of bio-fermented premix on the growth performance of beef cattle

[0157]

[0158] Note: Different letters marked for the data indicate significant differences.

[0159] 2.2 Effects of bio-fermented premix on the rumen fermentation parameters of beef cattle

[0160] As can be seen from the data in Table 4, the microbial protein content in the rumen of beef cattle in Group 2 and Group 3 was significantly increased by 56.92% and 42.31% respectively compared with the control group (P<0.05); the ammonia nitrogen content in the rumen of beef cattle in Group 2 and Group 3 was reduced by 11.29% and 9.83% respectively compared with the control group (P<0.05); the propionate content in the rumen of beef cattle in Group 2 and Group 3 was significantly increased by 18.58% and 19.07% respectively compared with the control group (P<0.05); the acetic acid / propionate value in the rumen of beef cattle in Group 2 and Group 3 was reduced by 13.08% and 13.08% respectively compared with the control group (P<0.05). It can be seen that the bio-fermented premix is superior to the control group and Group 1 in promoting the growth of rumen microbial protein, and the reduction of rumen ammonia nitrogen content indicates that the bio-fermented premix is more conducive to the utilization of ammonia nitrogen. At the same time, the bio-fermented premix also changes the fermentation type, promotes the growth of propionate-type fermentation strains, and is conducive to the weight gain of beef cattle.

[0161] Table 4 Effects of bio-fermented premix on rumen fermentation parameters of beef cattle

[0162]

[0163]

[0164] 2.3. Effects of bio-fermented premix on immune indexes of beef cattle

[0165] 2.3.1. Effects on immunoglobulin content

[0166] As can be seen from Table 5, the levels of immunoglobulin A in the serum of beef cattle in Group 2 and Group 3 were increased by 39.19% and 31.08% respectively compared with the control group (P<0.05); the levels of immunoglobulin G were significantly increased by 16.69% and 15.18% respectively compared with the control group (P<0.05).

[0167] Table 5 Effects of bio-fermented premix on immunoglobulin content of beef cattle (g / L)

[0168] Index Control group Group 1 Group 2 Group 3 Standard error P value Immunoglobulin A <![CDATA[0.74 b > <![CDATA[0.89 ab > <![CDATA[1.03 a > <![CDATA[0.97 a > 0.08 0.039 Immunoglobulin M 2.18 2.45 2.62 2.53 0.14 0.472 Immunoglobulin G <![CDATA[7.31 b > <![CDATA[7.92 ab > <![CDATA[8.53 a > <![CDATA[8.42 a > 0.46 0.048

[0169] 2.3.2. Effects on immune cytokine levels

[0170] The data in Table 6 show that the levels of interleukin-1 in the serum of beef cattle in Group 2 and Group 3 were significantly increased by 21.36% and 31.07% respectively (P<0.05), and the level of interleukin-4 in the serum of beef cattle in Group 2 was increased by 34.69% compared with the control group (P<0.05). It can be seen that the levels of interleukin-1 in the serum of Group 2 and Group 3 were increased, and at the same time, the content of interleukin-4 in the serum of beef cattle in Group 2 was also increased, indicating that the bio-fermented premix not only has a significant effect on changing the rumen fermentation type and improving feed digestibility, but also plays a good role in enhancing the immunity of beef cattle.

[0171] Table 6 Effects of Bio-Fermented Premix on the Levels of Immune Cytokines in Beef Cattle (ng / L)

[0172] Index Control group Group 1 Group 2 Group 3 Standard error P value Interleukin-1 <![CDATA[103.32 a > <![CDATA[116.06 ab > <![CDATA[125.17 b > <![CDATA[135.58 b > 5.06 0.041 Interleukin-2 118.34 125.04 131.36 127.52 4.42 0.153 Interleukin-4 <![CDATA[46.32 b > <![CDATA[50.58 ab > <![CDATA[62.39 a > <![CDATA[53.37 ab > 2.95 0.038 Interleukin-6 83.18 81.76 80.58 79.94 4.05 0.183 Interleukin-10 164.28 175.14 186.09 176.59 6.94 0.088 Tumor necrosis factor-α 326.45 318.74 314.05 306.82 14.39 0.418

[0173] 2.4. Effects of Bio-Fermented Premix on the Nutrient Digestibility of Beef Cattle

[0174] As can be seen from the data in Table 7, compared with the control group, the digestibility of dry matter in beef cattle in groups 2 and 3 increased significantly by 7.51% and 8.22% (P < 0.05); the digestibility of crude protein in beef cattle in all bio-fermented premix groups increased significantly by 5.01%, 6.21%, and 9.51% (P < 0.05); the digestibility of neutral detergent fiber in beef cattle in groups 2 and 3 increased by 5.93% and 7.38% (P < 0.05); the digestibility of acid detergent fiber in beef cattle in groups 2 and 3 increased by 9.58% and 8.53% (P < 0.05).

[0175] Table 7 Effects of Bio-Fermented Premix on the Nutrient Digestibility of Beef Cattle (%)

[0176] Index Control group Group 1 Group 2 Group 3 Standard error P value Dry matter <![CDATA[64.87 b > <![CDATA[67.63 ab > <![CDATA[69.74 a > <![CDATA[70.20 a > 4.13 0.018 Crude protein <![CDATA[56.65 b > <![CDATA[59.49 a > <![CDATA[60.17 a > <![CDATA[62.04 a > 3.84 0.014 Crude fat 72.31 72.83 73.52 74.27 5.05 0.241 Neutral detergent fiber <![CDATA[52.32 b > <![CDATA[53.85 ab > <![CDATA[55.42 a > <![CDATA[56.18 a > 3.58 0.046 Acid detergent fiber <![CDATA[50.53 b > <![CDATA[52.05 ab > <![CDATA[55.37 a > <![CDATA[54.84 a > 1.95 0.028 Calcium 48.47 49.05 49.62 50.41 3.04 0.527 Phosphorus 55.32 56.07 56.37 56.21 4.05 0.419

[0177] The embodiments described above are only descriptions of the preferred modes of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A bio-fermented premix for beef cattle, characterized in that, It comprises the following components in parts by mass: 20 - 40 parts of corn flour, 10 - 20 parts of soybean meal, 5 - 10 parts of wheat bran, 8 - 20 parts of grape seed extract, 3 - 10 parts of wheat germ, 2 - 10 parts of distillers' grains, 3 - 4 parts of parsley essential oil, 2 - 3 parts of eucommia ulmoides essential oil, 5 - 10 parts of fermentation product and 1 - 5 parts of trace element premix; The preparation method of the fermentation product includes the step of fermenting and culturing fermentation strains to obtain the fermentation product; the fermentation strains include Bacillus subtilis, lactic acid bacteria, yeast and Streptomyces fusidicogenes.

2. The bio-fermented premix according to claim 1, wherein The bio - fermented premix comprises the following components in parts by mass: 30 parts of corn flour, 15 parts of soybean meal, 8 parts of wheat bran, 15 parts of grape seed extract, 7 parts of wheat germ, 6 parts of distillers' grains, 3 parts of parsley essential oil, 3 parts of eucommia ulmoides essential oil, 8 parts of fermentation product and 3 parts of trace element premix.

3. The bio-fermented premix according to claim 1, characterized in that, The temperature of the fermentation culture is 28 - 42 °C, the time is 64 - 72 h, the initial pH value is 5.7 - 6.8, the moisture content is 37% - 40%, and the inoculation amount is 1% - 2%.

4. The bio-fermented premix according to claim 1, wherein The trace element premix comprises the following components: 50 - 200 mg / kg of copper, 100 - 2000 mg / kg of iron, 500 - 3000 mg / kg of zinc and 50 - 300 mg / kg of selenium.

5. The preparation method of the bio-fermented premix according to claim 1, characterized in that, It includes the following steps: Mix the corn flour, the soybean meal, the wheat bran, the grape seed extract, the wheat germ and the distillers' grains evenly to obtain a basic feed; Mix the parsley essential oil and the eucommia ulmoides essential oil evenly to obtain an essential oil combination; Ferment and culture the fermentation strains to obtain the fermentation product; the fermentation strains include Bacillus subtilis, lactic acid bacteria, yeast and Streptomyces fusidicogenes; Mix the basic feed, the essential oil combination, the fermentation product and the trace element premix to obtain the bio - fermented premix.

6. The preparation method according to claim 5, characterized in that, The temperature of the fermentation culture is 28 - 42 °C, the time is 64 - 72 h, the initial pH value is 5.7 - 6.8, the moisture content is 37% - 40%, and the inoculation amount is 1% - 2%.

7. Use of the bio - fermented premix according to any one of claims 1 - 4 in the preparation of a feed additive or feed for beef cattle.

8. A feed additive for beef cattle, characterized in that, The feed additive comprises the bio - fermented premix according to any one of claims 1 - 4.

9. A feed for beef cattle, characterized in that, The feed comprises the bio - fermented premix according to any one of claims 1 - 4.

10. Use of the bio - fermented premix according to any one of claims 1 - 4, the feed additive according to claim 8 or the feed according to claim 9 in any one or more of the following: (1) Application in improving the growth of beef cattle; (2) Application in alleviating rumen acidosis in beef cattle; (3) Application in enhancing the immunity of beef cattle; (4) Application in improving the digestibility of feed nutrients by beef cattle.

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