CN111000033A - Bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicine, preparation process and application - Google Patents
Bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicine, preparation process and application Download PDFInfo
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Abstract
The invention discloses a bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicines, a preparation process and application, wherein the preparation comprises charred triplet, bean pulp and thyme oil; the preparation is prepared by fermenting a mixture consisting of charred triplet and bean pulp through domesticated bacillus subtilis and then uniformly mixing the fermented mixture with thyme oil. The preparation process comprises the following steps: b, domesticating the bacillus subtilis: subculturing the bacillus subtilis by using a bacillus subtilis culture medium containing thyme oil to obtain a domesticated bacterial liquid; fermentation: sterilizing a mixture of charred triplet and bean pulp, uniformly mixing a sterilized bacillus subtilis culture medium containing thyme oil with the mixture, adding domesticated bacteria liquid, and performing closed culture to obtain a fermentation product; and (3) uniformly mixing the fermented product with thyme oil to obtain the oil-bacterium compound. The prepared oleobacter composite not only can fully exert the health-care and epidemic-prevention functions, but also is particularly suitable for raising poultry and livestock.
Description
Technical Field
The invention belongs to the technical field of feed additives, and particularly relates to a bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicines, and a preparation process and application thereof.
Background
Thyme oil has antibacterial, antiviral, antiparasitic, antispasmodic, and antioxidant effects. Research shows that the thyme oil has obvious inhibition effect on staphylococcus aureus, bacillus subtilis, escherichia coli, salmonella, listeria monocytogenes, aspergillus parasiticus and the like. Research shows that thyme oil has strong oxidation resistance and higher oxidation resistance than vitamin E. Through the effects of the functions of resisting oxidation, regulating immunity, improving intestinal flora and the like on animals, the thyme oil can effectively improve the production performance of the animals; however, when the growth promoter is used as the growth promoter, the growth promoter has no statistical significance on the influence difference of daily gain, daily feed intake and feed conversion ratio of livestock and poultry.
The bacillus subtilis is widely applied to aspects of livestock production and the like as a safe and effective probiotic, and is recorded in 'feed additive variety catalog' in 2013, and practice proves that the bacillus subtilis has definite functions of immune health care and promotion of animal growth, and is mainly reflected in that the bacillus subtilis plays a role in resisting bacteria and infection through space occupation competition, generation of antibacterial substances, bacteriolysis, biological oxygen deprivation and the like, and meanwhile, the bacillus subtilis also can enhance the immunity of livestock and poultry, promote the growth of other probiotics in the digestive tract, promote digestive absorption, supplement various vitamins and the like.
Thyme oil and bacillus subtilis are two different feed additive varieties, but have similar or complementary places in view of the functions in feed. With the release and implementation of the policy of limiting resistance and reducing resistance, antibacterial drugs are gradually quitting the team of feed additives, and the health care requirements under the condition of highly concentrated feeding are not reduced at all. In practical application, thyme oil and bacillus subtilis can partially meet the requirements of high intensification of animal husbandry, namely, the yield is improved, health care is realized, and the requirements of nature and no pollution are met, but the disadvantages exist, and the growth promotion and health care effects are difficult to achieve and the antibacterial drug level is difficult. Therefore, it is necessary to search for a composition of corresponding varieties or related products from natural products that have been primarily put into use at present.
In the prior art, thyme oil, as a component with an antibacterial function, has an obvious inhibiting effect on bacillus subtilis because of adverse effects on the survival of live bacillus subtilis. However, as for the functional direction of promoting animal production, the two have excellent effects on animal growth and production respectively, but because of the conflict between the two, no feed additive has the functions and effects of thyme oil and bacillus subtilis at present.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicines, a preparation process and application.
The technical scheme adopted by the invention is as follows:
a Bacillus subtilis viable bacteria fermented Chinese medicinal preparation containing thyme oil and Chinese medicinal materials comprises charred triplet, soybean meal and thyme oil; the preparation is prepared by fermenting a mixture consisting of charred triplet and bean pulp through domesticated bacillus subtilis and then uniformly mixing the fermented mixture with thyme oil.
Further, the weight ratio of the charred triplet to the bean pulp to the thyme oil is 90-99: 1-10: 2-20.
Furthermore, the domesticated bacillus subtilis is obtained by subculturing a culture medium added with essential oil inclusion powder, the essential oil inclusion powder is prepared from thyme oil and β -cyclodextrin, and the domesticated bacillus subtilis is domesticated bacillus subtilis I-X obtained by subculturing.
A preparation process of a bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicines comprises the following steps:
b, domesticating the bacillus subtilis: subculturing the bacillus subtilis by using a bacillus subtilis culture medium containing thyme oil to obtain a domesticated bacterial liquid;
fermentation: sterilizing a mixture of charred triplet and bean pulp, uniformly mixing a sterilized bacillus subtilis culture medium containing thyme oil with the mixture, adding domesticated bacteria liquid, and performing closed culture to obtain a fermentation product; and (3) uniformly mixing the fermented product with thyme oil to obtain the oil-bacterium compound.
Further, the domesticated bacterial liquid is an I-X generation domesticated bacterial liquid.
The preparation method of the thyme oil-containing bacillus subtilis culture medium comprises the following steps of dissolving β -cyclodextrin in water, adjusting the pH value to 8-11, adding thyme oil at 40-100 ℃ while stirring until oil drops appear on the liquid surface, adjusting the pH value to 4-7, filtering, and drying to obtain the thyme oil inclusion powder.
Further, the specific steps for obtaining any generation of domesticated bacterial liquid in the I generation to the X generation are as follows: dissolving the bacillus subtilis culture medium and the essential oil inclusion powder in water, adjusting the pH value to 6.5, sterilizing, cooling to room temperature, adding the previous generation bacillus subtilis liquid, shaking, and culturing at 28-37 ℃ to obtain the next generation domesticated bacteria liquid.
Further, the fermentation step comprises the following specific steps: mixing charred triplet and soybean meal, and sterilizing to obtain a first mixture; adding water into the bacillus subtilis culture medium and the essential oil inclusion powder, uniformly mixing and sterilizing to obtain a second mixture; and mixing the first mixture and the second mixture, adding domesticated bacteria liquid at room temperature, sealing, culturing at 28-37 ℃, drying in the shade, adding thyme oil, and uniformly mixing to obtain the oil-bacterium compound.
Further, when the previous generation of bacillus subtilis solution is the bacillus subtilis solution, the next generation of bacillus subtilis solution is the first generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium in the domestication step to the essential oil inclusion powder is 60.5: 0.5; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 99:1:2:30: 0.2;
when the previous generation of bacillus subtilis liquid is the first generation of bacillus subtilis liquid, the next generation of bacillus subtilis liquid is the second generation of bacillus subtilis liquid, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.4; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, bacillus subtilis culture medium and the essential oil inclusion powder is 98:2:4:30: 0.4;
when the previous generation of bacillus subtilis liquid is the second generation of bacillus subtilis liquid, the next generation of bacillus subtilis liquid is the third generation of bacillus subtilis liquid, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.6; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 97:3:6:30: 0.6;
when the previous generation of bacillus subtilis solution is the third generation of bacillus subtilis solution, the next generation of bacillus subtilis solution is the fourth generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.5; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 96:4:8:30: 0.8;
when the previous generation of bacillus subtilis solution is the IV generation of bacillus subtilis solution, the next generation of bacillus subtilis solution is the V generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.8; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 95:5:10:30: 1.0;
when the previous generation of bacillus subtilis solution is the V-th generation of bacillus subtilis solution, the next generation of bacillus subtilis solution is the VI-th generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.7; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 94:6:12:30: 1.2;
when the previous generation of bacillus subtilis solution is the bacillus subtilis solution of the VI generation, the next generation of bacillus subtilis solution is the bacillus subtilis solution of the VII generation, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 1; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 93:7:14:30: 1.4;
when the previous generation of bacillus subtilis solution is the VII-generation bacillus subtilis solution, the next generation of bacillus subtilis solution is the VIII-generation bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.9; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 92:8:16:30: 1.6;
when the previous generation of bacillus subtilis solution is the VIII generation of bacillus subtilis solution, the next generation of bacillus subtilis solution is the IX generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the acclimation step is 60.5: 1.2; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 91:9:18:30: 1.8;
when the previous generation of bacillus subtilis solution is the IX-generation bacillus subtilis solution, the next generation of bacillus subtilis solution is the X-generation bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 1.1; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 90:10:20:30: 2.0.
Use of a live bacterial preparation in the preparation of a feed additive.
The invention has the beneficial effects that: the method comprises the steps of adding trace amount of thyme oil into a bacillus subtilis culture medium, carrying out passage acclimation on bacillus subtilis, fermenting a mixture of scorched malt, scorched medicated leaven, scorched hawthorn fruit (the three substances are collectively called scorched triplet) and soybean meal by using the acclimated bacillus subtilis, and mixing the fermented scorched triplet and the soybean meal with a proper amount of thyme oil to obtain the thyme oil. The composition does not show adverse effects on each other to reduce the functions of the two parties, but shows high functional synergy; meanwhile, the preservation period of the bacillus subtilis is greatly prolonged. Is used for promoting the growth and production of animals, and the performance of the feed is far higher than that of thyme oil or bacillus subtilis which is used independently; the feed additive prepared by the viable bacteria preparation can obviously improve the immunity level of livestock, and particularly for pigs, the feed additive has better effects on the percentage of neutrophils in the blood of piglets, the antibody level of piglet plague and the lymphocyte transformation rate of piglets and promotes the growth of the piglets than astragalus polysaccharide. Therefore, the prepared oleobacter composite not only can fully exert the health-care and epidemic-prevention functions, but also is particularly suitable for feeding poultry (such as chickens, ducks and geese) and livestock (such as pigs, cows, sheep, horses and rabbits).
Detailed Description
The present invention is further illustrated below with reference to specific examples.
The next generation and the previous generation in the invention are used for representing two adjacent generations of domesticated bacillus subtilis liquid; if the previous generation of the third generation acclimated bacillus subtilis liquid is the second generation acclimated bacillus subtilis liquid, and so on.
Example 1
Materials β -Cyclodextrin (Zhiyuan Biotech, Inc. of Binzhou, Shandong), thyme oil (Jiangxin Xinsen, Inc. of Natural vegetable oils), and sodium citrate (commercially available).
β -cyclodextrin (1-30 parts) is taken, water (100 parts) is added and heated to be dissolved, sodium citrate is added to adjust the pH value to (8-11), the temperature is kept to (40-100 ℃), thyme oil (0.5-30 parts/min) is slowly added while stirring (60-600 revolutions/min) until oil drops appear on the liquid surface, hydrochloric acid is added to adjust the pH value to (4-7), the mixture is cooled to room temperature, filtering is carried out, drying is carried out at 40-60 ℃, and thyme oil β -cyclodextrin inclusion powder (essential oil inclusion powder for short) is obtained, wherein the content of thyme oil is 55%.
Example 2
Materials: standard bacillus subtilis strains (China general microbiological culture Collection center); the inclusion powder of essential oil (prepared in example 1), charred triplet (prepared by mixing equal parts of charred malt, charred medicated leaven and charred hawthorn, and the products of charred malt, charred medicated leaven and charred hawthorn, all of which are from Sichuan Koran Chinese herbal pieces of medicine, Inc.), and others are all commercially available products.
The bacillus subtilis culture medium: the beef extract consists of 20 parts of glucose, 15 parts of peptone, 5 parts of sodium chloride, 0.5 part of beef extract and 20 parts of agar.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 1000 parts of water to dissolve the bacillus subtilis culture medium, adjusting the pH value to 6.5, sterilizing for 15min at 121 ℃, cooling to room temperature, adding one bacillus subtilis frozen strain, shaking at 60-100 r/min, and culturing for 24-48 h at 28-37 ℃ to obtain an activated bacterial liquid. Taking out and storing at 4-6 ℃ for later use.
Taking 94 parts of charred triplet and 4 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium, 0.2 part of essential oil inclusion powder and 50 parts of water are taken and boiled for 30 minutes; mixing the two substances while the two substances are hot, cooling the two substances to room temperature, adding 20 parts of activated bacterium liquid, sealing, culturing for 24-48 h at 28-37 ℃, drying in the air at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 meshes of powder, measuring that the content of bacillus subtilis is more than or equal to 200 hundred million/g, adding 10 parts of essential oil blending powder, and uniformly mixing to obtain the oil bacterium composite reference substance.
Example 3
Materials: same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 0.5 part of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding one bacillus subtilis frozen strain, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain the first generation domesticated bacterial liquid. Taking out and storing at 4-6 ℃ for later use.
Taking 99 parts of charred triplet and 1 part of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium, 0.2 part of essential oil inclusion powder and 50 parts of water are taken and boiled for 30 minutes; mixing the two materials while the materials are hot, cooling the two materials to room temperature, adding 20 parts of the first generation domesticated bacteria liquid, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, measuring that the content of bacillus subtilis is more than or equal to 200 hundred million/g, adding 2 parts of thyme oil, and uniformly mixing to obtain the oil bacteria compound I.
Example 4
Materials: the first generation of acclimatized bacterial liquid (prepared in example 3); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 0.4 part of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of first generation domesticated bacterium liquid, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain second generation domesticated bacterium liquid. Taking out and storing at 4-6 ℃ for later use.
Taking 98 parts of charred triplet and 2 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 0.4 part of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two substances while the two substances are hot, cooling the two substances to room temperature, adding 20 parts of second generation domesticated bacteria liquid, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, measuring that the content of bacillus subtilis is more than or equal to 200 hundred million/g, adding 4 parts of thyme oil, and uniformly mixing to obtain an oil bacteria compound II.
Example 5
Materials: generation ii of acclimatized bacteria solution (prepared in example 4); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 0.6 part of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of the second generation domesticated bacterium liquid, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain the third generation domesticated bacterium liquid. Taking out and storing at 4-6 ℃ for later use.
97 parts of charred triplet and 3 parts of soybean meal are taken, crushed into 10-mesh powder, mixed evenly and sterilized by steam for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 0.6 part of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two materials while the materials are hot, cooling the two materials to room temperature, adding 20 parts of third generation domesticated bacteria liquid, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, measuring that the content of bacillus subtilis is more than or equal to 200 hundred million/g, adding 6 parts of thyme oil, and uniformly mixing to obtain an oil bacteria compound III.
Example 6
Materials: third generation acclimatized bacterial suspension (prepared in example 5); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 0.5 part of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of third generation domesticated bacterium liquid, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain fourth generation domesticated bacterium liquid. Taking out and storing at 4-6 ℃ for later use.
Taking 96 parts of charred triplet and 4 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 0.8 part of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two materials while the materials are hot, cooling the two materials to room temperature, adding 20 parts of IV generation domesticated bacteria liquid, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, measuring that the content of bacillus subtilis is more than or equal to 200 hundred million/g, adding 8 parts of thyme oil, and uniformly mixing to obtain an oil bacteria compound IV.
Example 7
Materials: the fourth generation of acclimatized bacterial liquid (prepared in example 6); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 0.8 part of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of an IV generation domesticated bacterium solution, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain a V generation domesticated bacterium solution. Taking out and storing at 4-6 ℃ for later use.
Taking 95 parts of charred triplet and 5 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 1.0 part of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two materials when the materials are hot, cooling the two materials to room temperature, adding 20 parts of a V-generation domesticated bacterium solution, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, measuring that the content of bacillus subtilis is more than or equal to 200 hundred million/g, adding 10 parts of thyme oil, and uniformly mixing to obtain an oil bacterium compound V.
Example 8
Materials: generation v of acclimatized bacterial suspension (prepared in example 7); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 0.7 part of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of a V-th generation domesticated bacterium solution, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain the VI-th generation domesticated bacterium solution. Taking out and storing at 4-6 ℃ for later use.
Taking 94 parts of charred triplet and 6 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 1.2 parts of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two materials when the materials are hot, cooling the two materials to room temperature, adding 20 parts of VI-generation domesticated bacteria liquid, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 meshes of powder, measuring that the bacillus subtilis is more than or equal to 200 hundred million/g, adding 12 parts of essential oil blending powder, and uniformly mixing to obtain an oil bacteria compound VI.
Example 9
Materials: the VI generation of domesticated bacteria (example 8 preparation); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 1 part of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of VI-generation domesticated bacteria liquid, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain VII-generation domesticated bacteria liquid. Taking out and storing at 4-6 ℃ for later use.
Taking 93 parts of charred triplet and 7 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 1.4 parts of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two materials when the mixture is hot, cooling the mixture to room temperature, adding 20 parts of VII-generation domesticated bacteria liquid, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature is not higher than 55 ℃) to 40-60 mesh powder, determining that the bacillus subtilis is more than or equal to 200 hundred million/g, adding 14 parts of essential oil composite powder, and uniformly mixing to obtain the oily bacteria composite VII.
Example 10
Materials: a VII-th generation of acclimatized bacterial liquid (prepared in example 9); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 0.9 part of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of a VII-generation domesticated bacterium solution, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain the VIII-generation domesticated bacterium solution. Taking out and storing at 4-6 ℃ for later use.
Taking 92 parts of charred triplet and 8 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 1.6 parts of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two substances when the mixture is hot, cooling the mixture to room temperature, adding 20 parts of the domesticated bacteria liquid of the eighth generation, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, measuring that the bacillus subtilis is more than or equal to 200 hundred million/g, adding 16 parts of essential oil binding powder, and uniformly mixing to obtain the oil bacteria compound VIII.
Example 11
Materials: acclimatized bacteria solution of the eighth generation (prepared in example 10); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 1.2 parts of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of a VIII generation domesticated bacterium solution, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain an IX generation domesticated bacterium solution. Taking out and storing at 4-6 ℃ for later use.
Taking 91 parts of charred triplet and 9 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 1.8 parts of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two substances while the mixture is hot, cooling the mixture to room temperature, adding 20 parts of the IX generation domesticated bacteria liquid, sealing, culturing at 28-37 ℃ for 24-48 h, drying at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, determining that the bacillus subtilis is more than or equal to 200 hundred million/g, adding 18 parts of essential oil composite powder, and uniformly mixing to obtain the oily bacteria composite IX.
Example 12
Materials: the IX generation of acclimatized bacterial suspension (prepared in example 11); the rest is the same as in example 2.
Taking 60.5 parts of a bacillus subtilis culture medium, adding 1.1 parts of essential oil inclusion powder, adding 1000 parts of water to dissolve, adjusting the pH value to 6.5, sterilizing at 121 ℃ for 15min, cooling to room temperature, adding 10 parts of IX generation domesticated bacterium liquid, shaking at 60-100 r/min, and culturing at 28-37 ℃ for 24-48 h to obtain the X generation domesticated bacterium liquid. Taking out and storing at 4-6 ℃ for later use.
Taking 90 parts of charred triplet and 10 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium and 2.0 parts of essential oil inclusion powder are taken, 50 parts of water is added, and the mixture is boiled for 30 minutes; mixing the two materials while the materials are hot, cooling the two materials to room temperature, adding 20 parts of the X-generation domesticated bacteria liquid, sealing, culturing for 24-48 h at 28-37 ℃, airing at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, measuring that the bacillus subtilis is more than or equal to 200 hundred million/g, adding 20 parts of essential oil binding powder, and uniformly mixing to obtain the oil bacteria compound X.
TABLE 1 variation of the principal materials and their amounts used in examples 1-12
Example 13
Materials: 130 rats (Sichuan academy of medical sciences) were tested, half female and half male; cyclophosphamide (commercially available); composite control of oil bacteria (prepared in example 1); oil bacteria complex I-X (oil bacteria complex of 5-9 in example 13).
Rats were randomly divided into 13 groups (10 per group, half male and half female) in turn, blank, negative, positive, and test i-x groups. Wherein, the blank group is conventionally raised; cyclophosphamide was added to the negative control group; the positive control group is added with an oil-adding bacterium compound reference substance, and the test groups I to X are sequentially added with oil-adding bacterium compound numbers I to X.
In addition to the blank group, each group of rats was injected with cyclophosphamide 0.1ml/d per day in the abdominal cavity during the test period; after three days, 1% of the corresponding experimental products were added to the drinking water of each group of rats in sequence every day except for the blank group and the negative control group. After 7 days, spleen index, thymus index and immunocyte CD4+/CD8+ of each group of rats were measured, respectively, and the results are shown in Table 2.
TABLE 2 comparison of weights of spleen and thymus and their indices for rats in different experimental groups
| Group of | CD4+/CD8+ | Spleen index | Index of thymus |
| Blank group | 2.57 | 0.0027 | 0.0016 |
| Negative control group | 1.57 | 0.0024 | 0.0011 |
| Positive control group | 2.24 | 0.0025 | 0.0019 |
| Test group I | 2.24 | 0.0030 | 0.0020 |
| Test II group | 2.26 | 0.0029 | 0.0020 |
| Test group III | 2.40 | 0.0029 | 0.0023 |
| Test group IV | 2.58 | 0.0030 | 0.0021 |
| Test group V | 2.38 | 0.0027 | 0.0019 |
| Test VI group | 2.48 | 0.0027 | 0.0019 |
| Test group VII | 2.58 | 0.0030 | 0.0021 |
| Experiment group VIII | 2.34 | 0.0028 | 0.0021 |
| Test group IX | 2.20 | 0.0025 | 0.0018 |
| Test group X | 2.23 | 0.0026 | 0.0016 |
From the results in Table 2, the data of the groups III to VIII, whether spleen index, thymus index, or immunoregulatory effect, are significantly better than those of the positive control group and the negative control group.
Example 14
Materials: 130 test mice (Sichuan academy of medical sciences), male and female; composite control of oil bacteria (prepared in example 1); oil bacteria complex I-X (oil bacteria complex of 5-9 in example 13).
Rats were randomly divided into 13 groups (10 per group, half male and half female) in turn, blank, negative, positive, and test i-x groups. Wherein, the blank group is conventionally raised; cyclophosphamide was added to the negative control group; the positive control group is added with an oil-adding bacterium compound reference substance, and the test groups I to X are sequentially added with oil-adding bacterium compound numbers I to X.
Fasting body weights of mice in all groups were determined on the first day. From the next day, 1% of the corresponding experimental products were added to the mice drinking water of each group in sequence every day for 7 consecutive days, except for the blank group. The fasting body weights of the mice in the experimental group were measured on the morning of day 8, and the average daily gain was calculated in grams, and the results are shown in Table 3.
TABLE 3 average daily growth of mice
| Group of | Mean initial weight | Average final weight | Average daily gain | Average daily intake | Material to weight ratio |
| Blank group | 20.58 | 23.75 | 0.45 | 5.98 | 13.21 |
| Control group | 20.2 | 23.41 | 0.46 | 5.91 | 12.89 |
| Test group I | 20.23 | 23.84 | 0.52 | 6.57 | 12.74 |
| Test II group | 20.19 | 23.98 | 0.54 | 6.43 | 11.88 |
| Test group III | 19.8 | 24.06 | 0.61 | 5.66 | 9.30 |
| Test group IV | 20.42 | 24.61 | 0.60 | 5.72 | 9.56 |
| Test group V | 20.07 | 24.52 | 0.64 | 6.05 | 9.52 |
| Test VI group | 20.29 | 24.7 | 0.63 | 6.16 | 9.78 |
| Test group VII | 20.68 | 24.75 | 0.58 | 5.4 | 9.29 |
| Experiment group VIII | 20.02 | 24.04 | 0.57 | 5.38 | 9.37 |
| Test group IX | 20.45 | 24.12 | 0.52 | 6.24 | 11.90 |
| Test group X | 20.45 | 24.02 | 0.51 | 5.96 | 11.69 |
From the results in table 3, the average daily gain of the test group v is the highest, the material-to-weight ratio of the test group vii is the lowest, and the tests group iii to viii are obviously superior to the blank group and the control group by combining the results of the two aspects.
Example 15
Materials: 130 rats (Sichuan academy of medical sciences) were tested, half female and half male; cyclophosphamide (commercially available); composite control of oil bacteria (prepared in example 1; closed, stored at room temperature in dry and ventilated place for 6 months); oil bacteria complex I-X (oil bacteria complex of 5-9 in example 13; sealed and stored at room temperature in dry and ventilated place for 6 months).
Rats were randomly divided into 13 groups (10 per group, half male and half female) in turn, blank, negative, positive, and test i-x groups. Wherein, the blank group is conventionally raised; cyclophosphamide was added to the negative control group; the positive control group is added with an oil-adding bacterium compound reference substance, and the test groups I to X are sequentially added with oil-adding bacterium compound numbers I to X.
In addition to the blank group, each group of rats was injected with cyclophosphamide 0.1ml/d per day in the abdominal cavity during the test period; after three days, 1% of the corresponding experimental products were added to the drinking water of each group of rats in sequence every day except for the blank group and the negative control group. After 7 days, spleen index, thymus index and immunocyte CD4+/CD8+ of each group of rats were measured, respectively, and the results are shown in Table 4.
TABLE 4 comparison of weights of spleen and thymus and their indices for rats in different experimental groups
| Group of | CD4+/CD8+ | Spleen index | Index of thymus |
| Blank group | 2.33 | 0.0027 | 0.0021 |
| Negative control group | 1.28 | 0.0022 | 0.0015 |
| Positive control group | 2.16 | 0.002 | 0.0018 |
| Test group I | 2.08 | 0.0024 | 0.0019 |
| Test II group | 1.99 | 0.0023 | 0.0016 |
| Test group III | 2.13 | 0.0029 | 0.0023 |
| Test group IV | 2.46 | 0.0028 | 0.0025 |
| Test group V | 2.4 | 0.003 | 0.0022 |
| Test VI group | 2.19 | 0.0028 | 0.0021 |
| Test group VII | 2.22 | 0.0027 | 0.0023 |
| Experiment group VIII | 2.37 | 0.0026 | 0.0022 |
| Test group IX | 2.12 | 0.0022 | 0.002 |
| Test group X | 2.12 | 0.002 | 0.0017 |
From the results in table 4, after 3 months of normal temperature storage, the data of the experimental groups iii to viii, whether spleen index, thymus index, or immunoregulatory effect, are significantly better than those of the positive control group and the negative control group; the data for the tests I to II and the tests IX to X are similar to those for the negative control group. Therefore, the invention objects obtained by domestication and experiments in groups III to VIII have higher stability, and are beneficial to long-term storage of the invention products.
Example 16
Materials: 130 test mice (Sichuan academy of medical sciences), male and female; composite control of oil bacteria (prepared in example 1; closed, stored at room temperature in dry and ventilated place for 6 months); oil bacteria complex I-X (oil bacteria complex of 5-9 in example 13; sealed and stored at room temperature in dry and ventilated place for 6 months).
Rats were randomly divided into 13 groups (10 per group, half male and half female) in turn, blank, negative, positive, and test i-x groups. Wherein, the blank group is conventionally raised; cyclophosphamide was added to the negative control group; the positive control group is added with an oil-adding bacterium compound reference substance, and the test groups I to X are sequentially added with oil-adding bacterium compound numbers I to X.
Fasting body weights of mice in all groups were determined on the first day. From the next day, 1% of the corresponding experimental products were added to the mice drinking water of each group in sequence every day for 7 consecutive days, except for the blank group. The fasting body weights of the mice in the experimental group were measured on the morning of day 8, and the average daily gain was calculated in grams, and the results are shown in table 5.
TABLE 5 average daily growth of mice
| Group of | Mean initial weight | Average final weight | Average daily gain | Average daily intake | Material to weight ratio |
| Blank group | 20.4 | 23.71 | 0.47 | 5.88 | 12.44 |
| Control group | 21.74 | 25.17 | 0.49 | 5.88 | 12.00 |
| Test group I | 20.27 | 23.85 | 0.51 | 6.27 | 12.26 |
| Test II group | 20.24 | 24 | 0.54 | 6.43 | 11.97 |
| Test group III | 19.83 | 24.11 | 0.61 | 5.76 | 9.42 |
| Test group IV | 20.44 | 24.64 | 0.60 | 5.81 | 9.68 |
| Test group V | 20.08 | 24.56 | 0.64 | 6.15 | 9.61 |
| Test VI group | 20.33 | 24.71 | 0.63 | 6.21 | 9.92 |
| Test group VII | 20.73 | 24.77 | 0.58 | 5.45 | 9.44 |
| Experiment group VIII | 20.05 | 24.09 | 0.58 | 5.38 | 9.32 |
| Test group IX | 20.47 | 24.15 | 0.53 | 6.24 | 11.87 |
| Test group X | 20.46 | 24.07 | 0.52 | 5.96 | 11.56 |
From the results in table 3, after 3 months of normal temperature storage, the average daily gain and the material weight ratio in the data of the test groups iii to viii are obviously superior to those of the positive control group and the negative control group; the data for the tests I to II and the tests IX to X are similar to those for the negative control group. Therefore, the invention objects obtained by domestication and experiments in groups III to VIII have higher stability, and are beneficial to long-term storage of the invention products.
Example 17
Materials: acclimatized bacteria solution of the eighth generation (prepared in example 10); the rest is the same as in example 2.
Taking 92 parts of charred triplet and 8 parts of soybean meal, crushing into 10-mesh powder, uniformly mixing, and introducing steam for sterilization for 30 minutes; meanwhile, 30 parts of bacillus subtilis culture medium, 0.2 part of essential oil inclusion powder and 50 parts of water are taken and boiled for 30 minutes; mixing the two materials when the materials are hot, cooling the two materials to room temperature, adding 20 parts of the domesticated bacteria liquid of the eighth generation, sealing, culturing for 24-48 h at 28-37 ℃, drying in the air at 40-55 ℃, crushing (the temperature does not exceed 55 ℃) to 40-60 mesh powder, measuring that the bacillus subtilis is more than or equal to 200 hundred million/g, adding 8 parts of essential oil composite powder, and uniformly mixing to obtain the oil-bacteria composite experimental product.
Experimental example-Effect of the Oenomycetes complexes on the immune function and daily gain of piglets
Materials: three-element piglets with the similar weight at the age of 20 days are 3 litters (10 piglets per litter and a certain pig farm of adult city); astragalus polysaccharides (commercially available); an oleaginous bacteria complex (prepared in example 5); hog cholera vaccine (Liaoning Yikang bio-corporation); hog cholera antibody detection kit (Lanzhou veterinary research institute of Chinese academy of agricultural sciences).
The method comprises the following steps: 3 litters of three-element piglets with the age of 20 days are injected with 4 parts of hog cholera vaccine and are randomly divided into a control group, an astragalus polysaccharide group and an oil bacterium compound group, wherein each group has 10 piglets, and the piglets are weaned at the age of 28 days; feeding astragalus polysaccharide (with the addition of 200mg/kg) to the astragalus polysaccharide group, feeding the olea europaea compound (with the addition of 200mg/kg) to the olea europaea compound group and not feeding to the control group from 25 days old; collecting blood at 25, 40 and 50 days of age, and performing immunity index detection; fasting body weights were measured at 28-day and 38-day ages, respectively, and the results are shown in table 6.
TABLE 6 Effect of different supplements on the percentage of neutrophils in the blood of piglets%
| Age of day | Control group | Astragalus polysaccharides group | Oil bacteria complex group |
| 25 | 9.878 | 9.604 | 8.136 |
| 40 | 6.411 | 15.874 | 18.517 |
| 50 | 5.746 | 7.642 | 9.434 |
As can be seen from table 6, at 25 days of age, the difference in percentage of neutrophils between the astragalus polysaccharide group and the olea europaea compound group was not significant; at 40 days of age, the percentage of peripheral blood neutrophils in the astragalus polysaccharide group and the oil bacteria compound group is obviously higher than that in the control group. The oil bacteria compound group is higher than the astragalus polysaccharide group, and the difference is not obvious. At 50 days of age, the astragalus polysaccharide content is higher than that of the control group, and the oil bacteria compound group is obviously higher than that of the astragalus polysaccharide group.
TABLE 7 Effect of different additives on piglet Swine fever antibody levels
| Age of day | Control group | Astragalus polysaccharides group | Oil bacteria complex group |
| 25 | 7.87 | 7.7 | 8.05 |
| 40 | 6.79 | 8.93 | 9.22 |
| 50 | 7.94 | 9.01 | 10.16 |
As can be seen from Table 7, the levels of the antibody against classical swine fever in the Astragalus polysaccharides group and the Oenothera odorata complex group were significantly higher than those in the control group at ages of 40 and 50 days. At 40 days of age, there was no significant difference between the astragalus polysaccharide group and the oleaginous bacteria complex group. At 50 days of age, the oil bacteria complex group is significantly higher than the astragalus polysaccharides group. The result shows that the difference of the antibody levels of the swine fever of the piglets of the astragalus polysaccharide group and the piglets of the oil bacterium compound group is not obvious at the age of 25 days, but the antibody level of the swine fever of the piglet of the oil bacterium compound group is not only obviously higher than that of a control group, but also obviously higher than that of the astragalus polysaccharide group at the age of 40 days and 50 days along with the increase of the feeding days. Therefore, the astragalus polysaccharide and the oil bacterium compound have certain promotion effect on the immune function of piglets, but still have better immune enhancement effect after the oil bacterium compound is applied.
TABLE 8 Effect of different additives on the lymphocyte transformation efficiency of piglets
| Age of day | Control group | Astragalus polysaccharides group | Oil bacteria complex group |
| 25 | 33.863 | 33.55 | 33.018 |
| 40 | 38.137 | 41.689 | 42.867 |
| 50 | 42.86 | 46.239 | 48.373 |
As can be seen from Table 8, the difference between the astragalus polysaccharide group and the oil bacteria compound group and the control group is not significant when the piglet peripheral lymphocyte transformation rate is 25 days old; at the age of 40 days and 50 days, the oil bacteria compound group is remarkably higher than that of the control group, and the difference between the rest groups is not remarkable. The result shows that the astragalus polysaccharide and oil bacteria compound can improve the conversion rate of the lymphocyte of the piglet, thereby improving the immunologic function of the piglet and having the highest conversion rate of the oil bacteria compound group.
TABLE 9 Effect of different supplements on average daily gain of piglets
As can be seen from Table 9, the average daily gain of piglets in the group added with astragalus polysaccharide did not change significantly compared with the control group; the average daily gain of piglets added with the oil bacterium compound group is obviously improved compared with that of the other two groups.
In conclusion, the oil-bacterium compound is superior to astragalus polysaccharide no matter the influence on the neutrophil percentage in the blood of the piglets, the influence on the piglet plague antibody level and the piglet lymphocyte transformation rate, and the promotion of the growth of the piglets. Therefore, the oleobacter composite prepared by the process provided by the invention not only can fully exert the health-care and epidemic-prevention functions, but also can ensure the economic performance of the oleobacter composite when being used as a feed raw material, and is particularly suitable for feeding poultry (such as chickens, ducks and geese) and livestock (such as pigs, cows, sheep, horses and rabbits).
The present invention is not limited to the above-described alternative embodiments, and various other forms of products can be obtained by anyone in light of the present invention. The above detailed description should not be taken as limiting the scope of the invention, which is defined in the claims, and which the description is intended to be interpreted accordingly.
Claims (10)
1. A bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicines is characterized in that: the preparation comprises charred triplet, soybean meal and thyme oil; the preparation is prepared by fermenting a mixture consisting of charred triplet and bean pulp through domesticated bacillus subtilis and then uniformly mixing the fermented mixture with thyme oil.
2. The viable bacillus subtilis fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicines according to claim 1 is characterized in that: the weight ratio of the charred triplet to the bean pulp to the thyme oil is 90-99: 1-10: 2-20.
3. The viable bacillus subtilis fermentation traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicines is characterized in that the domesticated bacillus subtilis is obtained by subculturing in a culture medium added with essential oil inclusion powder, the essential oil inclusion powder is prepared from thyme oil and β -cyclodextrin, and the domesticated bacillus subtilis is domesticated bacillus subtilis in generations I-X after subculturing.
4. A process for preparing a live bacillus subtilis fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicine according to any one of claims 1 to 3, which is characterized in that: the method comprises the following steps:
b, domesticating the bacillus subtilis: subculturing the bacillus subtilis by using a bacillus subtilis culture medium containing thyme oil to obtain a domesticated bacterial liquid;
fermentation: sterilizing a mixture of charred triplet and bean pulp, uniformly mixing a sterilized bacillus subtilis culture medium containing thyme oil with the mixture, adding domesticated bacteria liquid, and performing closed culture to obtain a fermentation product; and (3) uniformly mixing the fermented product with thyme oil to obtain the oil-bacterium compound.
5. The preparation process of the viable bacillus subtilis fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicine according to claim 4 is characterized in that: the domesticated bacterial liquid is an I-X generation domesticated bacterial liquid.
6. The preparation process of the viable bacillus subtilis fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicines is characterized in that the bacillus subtilis culture medium containing thyme oil is prepared by mixing essential oil inclusion powder and a bacillus subtilis culture medium with water, and the essential oil inclusion powder is prepared by the steps of dissolving β -cyclodextrin in water, adjusting the pH value to 8-11, stirring and adding thyme oil at 40-100 ℃ until oil drops appear on the liquid surface, adjusting the pH value to 4-7, filtering and drying to obtain the essential oil inclusion powder.
7. The preparation process of the bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicine according to claim 6, is characterized in that: the specific steps of obtaining any generation of domesticated bacteria liquid in the I generation to the X generation are as follows: dissolving the bacillus subtilis culture medium and the essential oil inclusion powder in water, adjusting the pH value to 6.5, sterilizing, cooling to room temperature, adding the previous generation bacillus subtilis liquid, shaking, and culturing at 28-37 ℃ to obtain the next generation domesticated bacteria liquid.
8. The preparation process of the bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicine according to claim 7 is characterized in that: the fermentation step comprises the following specific steps: mixing charred triplet and soybean meal, and sterilizing to obtain a first mixture; adding water into the bacillus subtilis culture medium and the essential oil inclusion powder, uniformly mixing and sterilizing to obtain a second mixture; and mixing the first mixture and the second mixture, adding domesticated bacteria liquid at room temperature, sealing, culturing at 28-37 ℃, drying in the shade, adding thyme oil, and uniformly mixing to obtain the oil-bacterium compound.
9. The preparation process of the bacillus subtilis viable bacteria fermented traditional Chinese medicine preparation containing thyme oil and traditional Chinese medicine according to claim 8, is characterized in that:
when the previous generation of bacillus subtilis liquid is the bacillus subtilis liquid, the next generation of bacillus subtilis liquid is the first generation of bacillus subtilis liquid, and the weight ratio of the bacillus subtilis culture medium in the domestication step to the essential oil inclusion powder is 60.5: 0.5; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 99:1:2:30: 0.2;
when the previous generation of bacillus subtilis liquid is the first generation of bacillus subtilis liquid, the next generation of bacillus subtilis liquid is the second generation of bacillus subtilis liquid, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.4; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, bacillus subtilis culture medium and the essential oil inclusion powder is 98:2:4:30: 0.4;
when the previous generation of bacillus subtilis liquid is the second generation of bacillus subtilis liquid, the next generation of bacillus subtilis liquid is the third generation of bacillus subtilis liquid, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.6; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 97:3:6:30: 0.6;
when the previous generation of bacillus subtilis solution is the third generation of bacillus subtilis solution, the next generation of bacillus subtilis solution is the fourth generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.5; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 96:4:8:30: 0.8;
when the previous generation of bacillus subtilis solution is the IV generation of bacillus subtilis solution, the next generation of bacillus subtilis solution is the V generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.8; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 95:5:10:30: 1.0;
when the previous generation of bacillus subtilis solution is the V-th generation of bacillus subtilis solution, the next generation of bacillus subtilis solution is the VI-th generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.7; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 94:6:12:30: 1.2;
when the previous generation of bacillus subtilis solution is the bacillus subtilis solution of the VI generation, the next generation of bacillus subtilis solution is the bacillus subtilis solution of the VII generation, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 1; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 93:7:14:30: 1.4;
when the previous generation of bacillus subtilis solution is the VII-generation bacillus subtilis solution, the next generation of bacillus subtilis solution is the VIII-generation bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 0.9; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 92:8:16:30: 1.6;
when the previous generation of bacillus subtilis solution is the VIII generation of bacillus subtilis solution, the next generation of bacillus subtilis solution is the IX generation of bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the acclimation step is 60.5: 1.2; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 91:9:18:30: 1.8;
when the previous generation of bacillus subtilis solution is the IX-generation bacillus subtilis solution, the next generation of bacillus subtilis solution is the X-generation bacillus subtilis solution, and the weight ratio of the bacillus subtilis culture medium to the essential oil inclusion powder in the domesticating step is 60.5: 1.1; in the fermentation step, the weight ratio of charred triplet, bean pulp, thyme oil, the bacillus subtilis culture medium and the essential oil inclusion powder is 90:10:20:30: 2.0.
10. Use of a live bacterial preparation according to any of claims 1-4 for the preparation of a feed additive.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114601018A (en) * | 2022-03-30 | 2022-06-10 | 江西天佳生物工程股份有限公司 | Feed additive, preparation method and application thereof in preventing intestinal infection of livestock and poultry |
| CN114796310A (en) * | 2020-12-31 | 2022-07-29 | 广东丸美生物技术股份有限公司 | Pharmaceutical composition containing ATCC9080 strain fermentation product, and preparation method and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101684450A (en) * | 2008-09-22 | 2010-03-31 | 北京欣博阳科技有限公司 | Fodder premix and special bacillus subtilis thereof |
| CN101690743A (en) * | 2009-09-30 | 2010-04-07 | 北京大北农动物保健科技有限责任公司 | Traditional Chinese medicine biological agent for improving immunity of livestock and preparation method thereof |
| CN105432938A (en) * | 2014-08-29 | 2016-03-30 | 鹤庆县中宝种植养殖场 | Antimicrobial-free laying hen feed |
| CN107981049A (en) * | 2017-12-29 | 2018-05-04 | 广西壮族自治区畜牧研究所 | A kind of feed addictive for improving measurement techniques for quality detection of meat and preparation method thereof |
| CN108125021A (en) * | 2017-11-10 | 2018-06-08 | 湖北华大瑞尔科技有限公司 | A kind of feed additive for controlling diarrhea rate and preparation method thereof |
| US20180344795A1 (en) * | 2015-09-01 | 2018-12-06 | Delacon Biotechnik Gmbh | Poultry feed additive |
| CN110558429A (en) * | 2019-10-12 | 2019-12-13 | 四川生力源生物工程有限公司 | domestication method of astragalus domestication yeast, process for preparing astragalus crude extract, astragalus crude extract and application thereof |
-
2019
- 2019-12-27 CN CN201911380196.0A patent/CN111000033A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101684450A (en) * | 2008-09-22 | 2010-03-31 | 北京欣博阳科技有限公司 | Fodder premix and special bacillus subtilis thereof |
| CN101690743A (en) * | 2009-09-30 | 2010-04-07 | 北京大北农动物保健科技有限责任公司 | Traditional Chinese medicine biological agent for improving immunity of livestock and preparation method thereof |
| CN105432938A (en) * | 2014-08-29 | 2016-03-30 | 鹤庆县中宝种植养殖场 | Antimicrobial-free laying hen feed |
| US20180344795A1 (en) * | 2015-09-01 | 2018-12-06 | Delacon Biotechnik Gmbh | Poultry feed additive |
| CN108125021A (en) * | 2017-11-10 | 2018-06-08 | 湖北华大瑞尔科技有限公司 | A kind of feed additive for controlling diarrhea rate and preparation method thereof |
| CN107981049A (en) * | 2017-12-29 | 2018-05-04 | 广西壮族自治区畜牧研究所 | A kind of feed addictive for improving measurement techniques for quality detection of meat and preparation method thereof |
| CN110558429A (en) * | 2019-10-12 | 2019-12-13 | 四川生力源生物工程有限公司 | domestication method of astragalus domestication yeast, process for preparing astragalus crude extract, astragalus crude extract and application thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114796310A (en) * | 2020-12-31 | 2022-07-29 | 广东丸美生物技术股份有限公司 | Pharmaceutical composition containing ATCC9080 strain fermentation product, and preparation method and application thereof |
| CN114796310B (en) * | 2020-12-31 | 2023-04-28 | 广东丸美生物技术股份有限公司 | Pharmaceutical composition containing ATCC9080 strain fermentation product, and preparation method and application thereof |
| CN114601018A (en) * | 2022-03-30 | 2022-06-10 | 江西天佳生物工程股份有限公司 | Feed additive, preparation method and application thereof in preventing intestinal infection of livestock and poultry |
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