Fermentation medicine residue as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of waste treatment and resource utilization, and particularly relates to fermentation medicine residues and a preparation method and application thereof. According to the preparation method of the fermented decoction dregs, two wastes, namely yeast fermentation residual liquid and radix isatidis decoction dregs, are used as main substrates, and lactic acid bacteria and saccharomycetes are used for two-step fermentation to obtain the fermented decoction dregs. The fermented medicine residue prepared by the method is good in smell and also contains more viable probiotics, the crude protein content, the acid-soluble protein content and the free amino acid content are all increased, and the fermented medicine residue can be used as a feed additive. The fermented decoction dregs are added into the basal feed of the mutton sheep, so that the negative influence caused by transportation stress can be relieved, the diarrhea rate can be reduced, and the growth performance of the mutton sheep can be improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of waste treatment and resource utilization, and particularly relates to a fermented medicinal residue, a preparation method thereof, and an application thereof. Background Art

[0002] A large amount of fermentation residue liquid is produced during the yeast production process. The fermentation residue liquid is the fermented concentrated liquid remaining after yeast centrifugation extraction. It not only contains a large amount of non-sugar substances that cannot be metabolized and utilized by yeast, but also contains nutrients added during the fermentation process, yeast metabolites, and residual yeast cells, including organic substances such as sugars, yeast proteins, amino acids, vitamins, colloidal substances, and organic acids, as well as inorganic substances such as nitrogen, phosphorus, potassium, calcium, and magnesium. These organic and inorganic substances are non-toxic and harmless, belong to the essential nutrient elements for the growth of plants and microorganisms, are beneficial to the intestinal digestion and individual growth of animals, are recyclable resources, and have the potential to be applied in feeds. However, the yeast fermentation residue liquid has a relatively dark color, a viscous texture, and poor palatability. The direct feeding effect is not good and it is not easy to be used as a feed raw material. The current utilization methods mainly focus on preparing fertilizers after evaporation and drying, and the economic benefits of recycling are not high.

[0003] Traditional Chinese medicines also face the situation of low recycling rate. Banlangen is a traditional Chinese medicine in China, which has the effects of clearing heat and detoxifying, cooling blood and relieving sore throat. It is one of the commonly used medicinal materials for respiratory diseases and is widely used. Its annual output is more than 1 million tons. At the same time, the output of Banlangen medicinal residue is also high. After Banlangen is processed and extracted, a large amount of available components still remain in the medicinal residue, such as crude fiber, crude protein, reducing sugar, starch, minerals, flavonoids, organic acids, etc., which have certain feeding value. However, the medicinal residue is difficult to digest for livestock, resulting in the ineffective components being not easily absorbed and utilized, and the unique smell of the medicinal residue will also affect the feeding of animals. Therefore, the usage amount in feed raw materials is usually small. Therefore, the current treatment methods of Banlangen medicinal residue mainly include incineration, burial, and stacking, which cannot recycle these effective components and will also cause secondary pollution.

[0004] In China, the common fattening method for meat sheep is off-site fattening, and the purchase, sale and transportation of sheep are very frequent. During transportation, stress in sheep can lead to disorders of body metabolism and functions, easily causing various respiratory and digestive diseases and resulting in economic losses to varying degrees. Banlangen (Isatis indigotica Fort.) has a certain effect on respiratory diseases due to its functions of clearing heat and detoxifying, cooling blood and relieving sore throat. The anti-inflammatory and antibacterial components it is rich in can also play a role when the immunity of sheep is low during transportation stress. Then, the Banlangen residue containing the effective components of Banlangen may also play the same role in the transportation stress of sheep. In addition, improving the feed intake of meat sheep and promoting their growth are important links in livestock production. Increasing the weight gain of meat sheep can reduce the fattening time, lower costs such as feed, labor and site, and increase the income of farmers. The method of making feed additives by fermenting Banlangen residue may solve the problem of resource utilization of Banlangen residue and relieve the stress of meat sheep, and improve the growth performance of meat sheep. However, Banlangen residue is bitter, and the method of fermenting Banlangen residue has limited effect on improving the taste of Banlangen residue. After adding it to the feed, it will reduce the feed intake of sheep and is not easily accepted by sheep. Moreover, the content of effective components in Banlangen residue is small, and the active components in the fermented product may not be sufficient to relieve the stress of meat sheep and improve the growth performance of meat sheep. Therefore, existing research often uses multiple traditional Chinese medicines or residues for fermentation, and the proportion of Banlangen residue is small. Using this method to solve the problem of resource utilization of Banlangen residue has certain limitations. Summary of the Invention

[0005] In view of the above technical problems, the present invention provides a fermented residue and its preparation method and application. The fermented residue provided by the present invention is obtained by solid-state fermentation using yeast fermentation residue and Banlangen residue as the main substrates. It not only has a good smell, but also is rich in live probiotics, crude protein, acid-soluble protein and free amino acids, and can be used as a feed additive, which not only solves the problem of recycling of two kinds of waste, namely yeast fermentation residue and Banlangen residue, but also can relieve the stress of meat sheep and improve the growth performance of meat sheep.

[0006] To achieve the above invention purpose, the present invention adopts the following technical solutions: The first aspect of the present invention provides a preparation method of a fermented residue, which specifically includes the following steps: S1. Activate lactic acid bacteria and yeast and culture them to the logarithmic growth phase, and use them as lactic acid bacteria and yeast seed solutions respectively; dilute the yeast fermentation residue 5-8 times to obtain a diluted yeast fermentation residue solution; mix the diluted yeast fermentation residue solution, Banlangen residue and corncob powder according to the mass ratio of (1.5-9):2:(0.8-1.2), add urea accounting for 0.5%-3.0% of the mass of the obtained mixture to the mixture, and stir evenly to obtain a solid-state fermentation medium; S2. Inoculate the solid fermentation medium with the lactic acid bacteria seed liquid at an inoculation amount of 1% - 5%, stir evenly, ferment at 25 - 38°C for 1 - 5 days, and then inoculate a mixed seed liquid with a volume ratio of the lactic acid bacteria seed liquid to the yeast seed liquid of (1 - 3):(1 - 4) at an inoculation amount of 2% - 10%, stir evenly, and ferment at 25 - 38°C for 3 - 15 days.

[0007] The fermented medicinal residues prepared by this preparation method have a good smell, and at the same time also have a high viable count of probiotics, crude protein content, acid-soluble protein content and free amino acid content, as well as good antioxidant activity, and can be used as a feed additive. Experiments have proved that adding this fermented medicinal residue to the basic feed of meat sheep can significantly relieve the transportation stress of meat sheep, reduce the diarrhea rate, and improve the growth performance of meat sheep.

[0008] This preparation method uses yeast fermentation residue liquid and Isatis root medicinal residues as substrates, which can not only make full use of the residual nutrients in the waste, but also reduce environmental protection and production costs. The high-value products produced can also bring certain economic benefits to the enterprise. It not only provides a practical solution for the reuse of yeast fermentation residue liquid and Isatis root medicinal residues, but also provides a new idea for solving the transportation stress problem of meat sheep.

[0009] Preferably, the lactic acid bacteria in S1 are Enterococcus faecium.

[0010] Preferably, the yeast in S1 is Saccharomyces cerevisiae.

[0011] Preferably, the lactic acid bacteria and yeast in S1 are selected from strains that can grow well in a medium containing yeast fermentation residue liquid.

[0012] Further preferably, the screening method for the lactic acid bacteria and yeast is as follows: After activating the lactic acid bacteria and yeast to be screened, inoculate them into the yeast fermentation residue liquid medium at an inoculation amount of 3% for cultivation, select the strains with a high viable count for subculture, continue to cultivate the subcultured strains in the yeast fermentation residue liquid medium, repeat the operation until subcultured for 10 - 15 generations, and then select the strain with the highest viable count, which is the lactic acid bacteria and yeast in S1; the composition of the yeast fermentation residue liquid medium is: yeast fermentation residue liquid and water with a volume ratio of 1:4, and 10 g / L urea.

[0013] Preferably, the mass ratio of the yeast fermentation residue liquid dilution, Isatis root medicinal residues, and corncob powder in S1 is 6:2:1.

[0014] Preferably, the addition amount of urea in S1 is 1.0% - 2.0%. The further preferably addition amount is 1.5% - 2.0%. The even more preferably addition amount is 1.5%.

[0015] Preferably, the inoculation amount of the lactic acid bacteria seed liquid in S2 is 1% - 3%. The further preferably inoculation amount is 2%.

[0016] Preferably, the fermentation time after inoculating the lactic acid bacteria seed liquid in S2 is 2 - 4 days. The further preferably fermentation time is 3 days.

[0017] Preferably, the volume ratio of the lactic acid bacteria seed liquid to the yeast seed liquid in the mixed seed liquid in S2 is 1:(1 - 3). The further preferably volume ratio is 1:1.

[0018] Preferably, the inoculation amount of the mixed seed liquid in S2 is 6% - 8%. The further preferably inoculation amount is 8%.

[0019] Preferably, the fermentation time after inoculating the mixed seed liquid in S2 is 6 - 9 days. The further preferably fermentation time is 6 days.

[0020] Preferably, the temperature of the fermentation in S2 is 25 - 32 °C. The further preferably temperature is 30 °C.

[0021] The second aspect of the present invention provides a fermented medicinal residue prepared by the above preparation method.

[0022] The third aspect of the present invention provides the application of the above fermented medicinal residue as a feed additive.

[0023] Preferably, the feed additive is a sheep feed additive.

[0024] More preferably, the feed additive is a meat sheep feed additive.

[0025] The above fermented medicinal residue has the functions of relieving the transportation stress of meat sheep, reducing the diarrhea rate, and can also improve the growth performance of healthy meat sheep, ensuring the smooth progress of the fattening process, thereby effectively reducing economic losses and improving economic benefits.

[0026] The fourth aspect of the present invention provides a method for relieving the transportation stress of meat sheep and / or reducing the diarrhea rate: adding 5 - 10%wt of the above fermented medicinal residue to the meat sheep diet.

[0027] The fifth aspect of the present invention provides a method for improving the growth performance of meat sheep: adding 3% - 10%wt of the above fermented medicinal residue to the meat sheep diet.

[0028] The present invention does not limit the preparation method and source of yeast fermentation residue and radix isatidis medicinal residue. The yeast fermentation residue and radix isatidis medicinal residue obtained in conventional production are applicable to the present invention.

[0029] Exemplarily, the yeast fermentation residue can be prepared by the following method: using molasses as the main raw material, adding other nutrients as the culture medium, inoculating Saccharomyces cerevisiae for fermentation; after the fermentation is completed, the fermentation broth is separated by a centrifuge to obtain yeast mud and fermentation supernatant, and this supernatant is the yeast fermentation residue.

[0030] Exemplarily, the Isatis indigotica residue can be prepared by the following method: after the Isatis indigotica medicinal materials are washed, dried, and coarsely pulverized, they are mixed with deionized water in a ratio of 1:10, heated and decocted, and kept boiling for 3 hours. After that, the extract and the residue are separated. The residue is washed with an appropriate amount of deionized water to remove the residual liquid and then dried at 80 °C. After drying, it is pulverized again and passed through a 40-mesh sieve to obtain the product.

[0031] The beneficial effects of the present invention are as follows: The present invention establishes a method for solid-state fermentation with yeast fermentation residue and Isatis indigotica residue as the main substrates, adding probiotics to obtain a fermented residue that can relieve the transportation stress of meat sheep, reduce the diarrhea rate, and promote the growth of meat sheep, as well as its preparation method. It provides a practical solution for the reuse of yeast fermentation residue and Isatis indigotica residue, and at the same time provides a feasible product for relieving the transportation stress of meat sheep, reducing the diarrhea rate, promoting the growth of meat sheep, and reducing economic losses.

[0032] The preparation method provided by the present invention enables both lactic acid bacteria and yeast to reproduce and grow better through a two-stage inoculation method, and the residual nutrients in the yeast fermentation residue and Isatis indigotica residue can also be fully utilized, so that the fermented residue has richer metabolites and bioactive substances, thereby improving the effect of the residue. Experiments have proved that compared with other fermentation methods, the viable count of probiotics, the content of crude protein, acid-soluble protein, and free amino acids in the fermented residue prepared by this preparation method are significantly increased, and the smell is good and it is easily accepted by livestock such as meat sheep; compared with the solid medium before fermentation, the fermented residue obtained by fermenting with this preparation method also has higher antioxidant activity. Adding this fermented residue to the feed to feed meat sheep with transportation stress can relieve the negative effects brought by stress and reduce the diarrhea rate; feeding healthy meat sheep can increase the daily weight gain of meat sheep. Description of the Drawings Figure 1 It is the viable count of yeast in the fermented residue obtained by fermentation with different urea addition amounts in Test Example 3 of the present invention; Figure 2 It is the viable count of yeast in the fermented residue obtained by fermentation with different solid-liquid ratios and fermentation days in Test Example 4 of the present invention; Figure 3 It is the viable count of yeast in the fermented residue obtained by fermentation with different inoculation amounts of Enterococcus faecium seed liquid in Test Example 5 of the present invention; Figure 4It is the viable yeast count in the fermented medicinal residues obtained by fermentation at different fermentation days after inoculating Enterococcus faecium seed liquid in Test Example 6 of the present invention; Figure 5 It is the viable yeast count in the fermented medicinal residues obtained by fermentation using different mixed seed liquids in Test Example 7 of the present invention; Figure 6 It is the viable yeast count in the fermented medicinal residues obtained by fermentation with different inoculation amounts of mixed seed liquids in Test Example 8 of the present invention; Figure 7 It is the viable yeast count in the fermented medicinal residues obtained by fermentation at different fermentation temperatures in Test Example 9 of the present invention; Figure 8 It is the viable yeast count in the fermented medicinal residues obtained by fermentation at different fermentation days after inoculating the mixed seed liquid in Test Example 10 of the present invention. Detailed implementation manners

[0033] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts fall within the protection scope of the present invention.

[0034] The yeast fermentation residue liquid and the radix isatidis medicinal residues are rich in nutrients, and the total annual output is relatively large, but the actual proportion applied to animal production is not high. The method of fermenting traditional Chinese medicine medicinal residues and then making them into feed additives is expected to solve the problem of resource utilization of radix isatidis medicinal residues. However, due to the bitter taste of radix isatidis, the fermented product often has an unpleasant odor and is not easily accepted by animals. For example, sheep prefer sweet tastes, have an average preference for sour tastes, and although they have a stronger tolerance for bitter tastes than cattle, the bitter taste in the feed will still reduce the feed intake of sheep.

[0035] Based on the research on the fermentation method, the embodiment of the present invention provides a preparation method of fermented medicinal residues mainly using yeast fermentation residue liquid and radix isatidis medicinal residues, which specifically includes the following steps: S1. Activate lactic acid bacteria and yeast, and culture them to the logarithmic growth phase to obtain lactic acid bacteria and yeast seed liquids respectively; dilute the yeast fermentation residue liquid by 5 to 8 times to obtain a diluted yeast fermentation residue liquid; mix the diluted yeast fermentation residue liquid, radix isatidis medicinal residues, and corncob powder according to a mass ratio of (1.5 to 9):2:(0.8 to 1.2), add urea accounting for 0.5% to 3.0% of the mass of the obtained mixture, and stir evenly to obtain a solid-state fermentation medium; S2. Inoculate the solid fermentation medium with the lactic acid bacteria seed liquid at an inoculation amount of 1% - 5%, stir evenly, and ferment at 25 - 38°C for 1 - 5 days. Then, inoculate a mixed seed liquid with a volume ratio of the lactic acid bacteria seed liquid to the yeast seed liquid of (1 - 3):(1 - 4) at an inoculation amount of 2% - 10%, stir evenly, and ferment at 25 - 38°C for 3 - 15 days.

[0036] The fermented medicinal residues prepared by this preparation method have a good smell, and at the same time, also have a high viable yeast count, crude protein content, acid-soluble protein content, free amino acid content, and good antioxidant activity.

[0037] The embodiments of the present invention also provide fermented medicinal residues prepared according to the above preparation method.

[0038] The embodiments of the present invention also provide the application of the above fermented medicinal residues as feed additives.

[0039] The following will be further illustrated with specific examples.

[0040] The original Enterococcus faecium and the original Saccharomyces cerevisiae used in the following examples were provided by Beijing Xindayang Technology Development Co., Ltd.

[0041] The preparation process of the yeast fermentation residue liquid used in the following examples is as follows: inoculate a medium containing 20 g / L molasses, 20 g / L peptone, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium chloride, 0.5 g / L magnesium sulfate, and 5 g / L sodium chloride with Saccharomyces cerevisiae (provided by Beijing Xindayang Technology Development Co., Ltd.) and ferment for 72 hours. After the fermentation is completed, separate the fermentation broth by a centrifuge to obtain yeast mud and fermentation supernatant, and this supernatant is the yeast fermentation residue liquid.

[0042] The preparation process of the Isatis root medicinal residues used in the following examples is as follows: after the Isatis root medicinal materials are washed, dried, and coarsely pulverized, mix them with deionized water in a ratio of 1:10, heat and decoct, and keep boiling for 3 hours. After completion, separate the extract and the medicinal residues, wash the medicinal residues with an appropriate amount of deionized water to remove the residual liquid, and then dry them at 80°C. After drying, pulverize them again and pass through a 40-mesh sieve to obtain the product.

[0043] The components of the yeast fermentation residue liquid medium used in the following examples are: 200 mL of yeast fermentation residue liquid, 10 g of urea, and 800 mL of water.

[0044] Unless otherwise specified, other raw materials, reagents, drugs, or instruments used in the following examples are all conventional commercially available products obtained through commercial channels. Unless otherwise specified, the methods used in the following examples are all conventional methods in the art.

[0045] Example 1 This embodiment provides a fermented medicinal residue. The preparation method is as follows: 1. Domestication and activation of fermentation strains (1) Domestication of fermentation strains Domestication: After activating the original Enterococcus faecium and original Saccharomyces cerevisiae strains, inoculate them into the yeast fermentation residue medium at an inoculation amount of 3% for liquid shake flask culture. Each batch has 40 parallel flasks. When sampling and detecting the viable cell count at 48 h, select the shake flasks with a high viable cell count for subculture, and continue culturing and detecting the viable cell count. After subculture and domestication to the 10th generation, select the domesticated strain with the highest viable cell count and freeze it.

[0046] (2) Activation of fermentation strains Activation: Inoculate the frozen Enterococcus faecium and Saccharomyces cerevisiae strains into the yeast fermentation residue medium, and culture them at 30 °C and 150 r / min for 24 h until the strains reach the logarithmic growth phase. At this time, the OD of Enterococcus faecium 560 = 0.7, and the OD of yeast 600 = 1.2. Use them as the Enterococcus faecium seed liquid and yeast seed liquid respectively.

[0047] 2. Preparation of solid fermentation medium Dilute the yeast fermentation residue 5 times to obtain a diluted yeast fermentation residue solution. Mix the diluted yeast fermentation residue solution, Isatis root medicinal residue, and corncob powder according to a mass ratio of 6:2:1. Add 1.5% of the mass of the mixture of urea as a nitrogen source to the obtained mixture, and stir all the materials evenly to obtain the solid fermentation medium.

[0048] 3. Fermentation First, inoculate the Enterococcus faecium seed liquid into the solid fermentation medium at an inoculation amount of 2%. After stirring evenly, load it into a fermentation bag with a one-way air release valve and ferment at 30 °C for 3 days. Then, inoculate a mixed seed liquid with a volume ratio of 1:1 of the Enterococcus faecium seed liquid and yeast seed liquid at an inoculation amount of 8%. After stirring evenly, load it into a fermentation bag with a one-way air release valve and ferment at 30 °C for 6 days.

[0049] During the fermentation process, the initial pH value of the solid fermentation medium is 5.3. After fermenting with the Enterococcus faecium seed liquid for 3 days, the pH value drops to 4.8. After fermenting with the mixed seed liquid for 6 days, the pH value drops to 4.2.

[0050] Example 2 This embodiment provides a fermented medicinal residue. Its preparation method is basically the same as that of Example 1, except that the addition amount of urea in step 2 is 1.0%.

[0051] Example 3 This embodiment provides a kind of fermented medicinal residues. Its preparation method is basically the same as that of Embodiment 2, the difference being that in step 3, the Enterococcus faecium seed liquid is inoculated into the solid fermentation medium at an inoculation amount of 2% first, and after stirring evenly, it is filled into a fermentation bag and fermented at 30°C for 1 day. Then, a mixed seed liquid with a volume ratio of Enterococcus faecium seed liquid to yeast seed liquid of 1:1 is inoculated at an inoculation amount of 8%, and after stirring evenly, it is filled into a fermentation bag and fermented at 30°C for 8 days.

[0052] Examples 4 to 7 Examples 4 to 7 respectively provide a kind of fermented medicinal residues. Its preparation method is basically the same as that of Embodiment 1, the difference being that the addition amounts of urea in step 2 are 0.5%, 2.0%, 2.5%, and 3.0% respectively.

[0053] Examples 8 to 11 Examples 8 to 11 respectively provide a kind of fermented medicinal residues. Its preparation method is basically the same as that of Embodiment 1, the differences being that in step 2, the yeast fermentation residue diluent, the radix isatidis medicinal residues, and the corncob powder are mixed according to the mass ratios of 1.5:2:1, 3:2:1, 6:2:1, and 9:2:1 respectively, that is, the solid-liquid ratios are 2:1, 1:1, 1:2, and 1:3 respectively; in step 3, the fermentation time after inoculating the mixed seed liquid is 8 days.

[0054] Examples 12 to 15 Examples 12 to 15 respectively provide a kind of fermented medicinal residues. Its preparation method is basically the same as that of Embodiment 1, the difference being that the inoculation amounts of the Enterococcus faecium seed liquid in step 3 are 1%, 3%, 4%, and 5% respectively.

[0055] Examples 16 to 19 Examples 16 to 19 respectively provide a kind of fermented medicinal residues. Its preparation method is basically the same as that of Embodiment 1, the difference being that the fermentation times after inoculating the Enterococcus faecium seed liquid in step 3 are 1, 2, 4, and 5 days respectively.

[0056] Examples 20 to 24 Examples 20 to 24 respectively provide a kind of fermented medicinal residues. Its preparation method is basically the same as that of Embodiment 1, the difference being that the volume ratios of the Enterococcus faecium seed liquid to the yeast seed liquid in the mixed seed liquid of step 3 are 3:1, 2:1, 1:2, 1:3, and 1:4 respectively.

[0057] Examples 25 to 28 Examples 25 to 28 respectively provide a kind of fermented medicinal residues. Its preparation method is basically the same as that of Embodiment 1, the difference being that the inoculation amounts of the mixed seed liquid in step 3 are 2%, 4%, 6%, and 10% respectively.

[0058] Examples 29 to 33 Examples 29 to 33 respectively provide a kind of fermented medicinal residues. The preparation method is basically the same as that of Example 1, except that the fermentation temperatures in step 3 are 25, 27, 32, 35, and 38 °C respectively.

[0059] Examples 34 to 37 Examples 34 to 37 respectively provide a kind of fermented medicinal residues. The preparation method is basically the same as that of Example 1, except that the fermentation times after adding the mixed seed liquid in step 3 are 3, 9, 12, and 15 days respectively.

[0060] Example 38 This example provides a kind of fermented medicinal residues. The preparation method is basically the same as that of Example 1, except that the dilution multiple of the yeast fermentation residue liquid in step 2 is 8 times.

[0061] Example 39 This example provides a kind of fermented medicinal residues. The preparation method is basically the same as that of Example 1, except that the mass ratio of the yeast fermentation residue liquid dilution, Isatis root medicinal residues, and corncob powder in step 2 is 6:2:0.8.

[0062] Example 40 This example provides a kind of fermented medicinal residues. The preparation method is basically the same as that of Example 1, except that the mass ratio of the yeast fermentation residue liquid dilution, Isatis root medicinal residues, and corncob powder in step 2 is 6:2:1.2.

[0063] Comparative Example 1 This comparative example provides a kind of fermented medicinal residues. The preparation method is basically the same as that of Example 2, except that in step 2, an equal volume of water is used to replace the seed liquid and inoculated onto the solid fermentation medium.

[0064] Comparative Example 2 This comparative example provides a kind of fermented medicinal residues. The preparation method is the same as that of Example 2, except that in step 3, the Enterococcus faecium seed liquid is inoculated onto the solid fermentation medium at an inoculation amount of 10%, evenly stirred and then filled into a fermentation bag and fermented at 30 °C for 9 d.

[0065] Comparative Example 3 This comparative example provides a kind of fermented medicinal residues. The preparation method is the same as that of Example 2, except that in step 3, the yeast seed liquid is inoculated onto the solid fermentation medium at an inoculation amount of 10%, evenly stirred and then filled into a fermentation bag and fermented at 30 °C for 9 d.

[0066] Comparative Example 4 This comparative example provides a fermented medicinal residue. The preparation method is the same as that of Example 2, except that in step 3, the yeast seed liquid is inoculated onto the solid fermentation medium at an inoculation amount of 2% first, evenly stirred and then filled into a fermentation bag for fermentation at 30 °C for 1 d. Then, a mixed seed liquid with a volume ratio of 1:1 of the Enterococcus faecium seed liquid and the yeast seed liquid is inoculated at an inoculation amount of 8%, evenly stirred and then filled into a fermentation bag for fermentation at 30 °C for 8 d.

[0067] Comparative Example 5 The method is the same as that of Example 1, except that the solid fermentation medium in step 2 is a 2:1 mixture of radix isatidis medicinal residue and corncob powder.

[0068] Inspection Example 1 Prepare the Enterococcus faecium seed liquid and the yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2). Use the same batch of Enterococcus faecium seed liquid and yeast seed liquid to prepare the fermented medicinal residue according to steps 2 and 3 of Example 2, Example 3 and Comparative Examples 1-4 respectively. After the fermentation in step 3 is completed, measure the pH, viable yeast count, acid-soluble protein content, free amino acid content of the fermented medicinal residue, and evaluate the odor. The results are shown in Table 1.

[0069] Table 1 Fermentation results of Example 2, Example 3 and Comparative Examples 1-4

[0070] It can be seen from the results in Table 1 that the fermented medicinal residue prepared by the preparation method of Example 2 has a better odor, with a sweet taste and a faint wine smell, and the highest viable bacteria count, acid-soluble protein content and free amino acid content; the fermented medicinal residue prepared by Example 3 has a sour taste and a faint wine smell, which is not as good as the odor of Example 2, but the viable bacteria count, acid-soluble protein content and total free amino acid content are higher than those of each comparative example. The odor, taste, viable bacteria count, acid-soluble protein content and free amino acid content of the fermented medicinal residue prepared by the preparation methods of Comparative Examples 1-4 are all inferior to those of Example 2 and Example 3.

[0071] Inspection Example 2 Prepare the Enterococcus faecium seed liquid and the yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2). Use the same batch of Enterococcus faecium seed liquid and yeast seed liquid to prepare the fermented medicinal residue according to steps 2 and 3 of Example 1 and Comparative Example 5 respectively, and measure the viable yeast count of the obtained fermented medicinal residue and evaluate the odor. The results are shown in Table 2.

[0072] Table 2 Fermentation results of Example 1 and Comparative Example 5

[0073] Test Example 3 Prepare Enterococcus faecium seed liquid and yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2). Use the same batch of Enterococcus faecium seed liquid and yeast seed liquid to prepare fermented medicinal residues respectively according to Steps 2 and 3 in Example 1, Example 2, Example 4 - 7, and measure the viable yeast count of the fermented medicinal residues obtained by fermentation.

[0074] The results are as Figure 1 shown. The viable count increases with the increase of the urea addition amount. When the addition amount reaches 1.5%, the viable count reaches the peak value of 1.2×10 8 CFU / g. When a higher urea addition amount is used, the viable count decreases instead.

[0075] Test Example 4 Prepare Enterococcus faecium seed liquid and yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2). Use the same batch of Enterococcus faecium seed liquid and yeast seed liquid to prepare fermented medicinal residues respectively according to Steps 2 and 3 in Example 1, Example 8 - 11, and measure the viable yeast count on the 2nd, 4th, 6th, and 8th days after inoculating the mixed seed liquid and starting fermentation.

[0076] The results are as Figure 2 shown. When the solid - liquid mass ratio is 1:2 (Example 1), the change in the viable count is the most significant, and it gradually increases with the extension of the fermentation time. When fermenting with yeast for 6 days, the viable count reaches the peak value of 11.8×10 7 CFU / g. Continuing to ferment until the 8th day, the viable count decreases. The change in the viable count under other solid - liquid ratios is not significant, and it even shows a gradually decreasing trend when the solid - liquid ratio is 2:1.

[0077] Test Example 5 Prepare Enterococcus faecium seed liquid and yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2). Use the same batch of Enterococcus faecium seed liquid and yeast seed liquid to prepare fermented medicinal residues respectively according to Steps 2 and 3 in Example 1, Example 12 - 15, and measure the viable yeast count of the fermented medicinal residues obtained by fermentation.

[0078] The results are as Figure 3 shown. When the inoculation amount of Enterococcus faecium seed liquid is 1% - 3%, the viable yeast count is relatively high. However, when the inoculation amount increases, the viable yeast count is lower.

[0079] Test Example 6 Prepare Enterococcus faecium seed liquid and yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2), and prepare fermented medicinal residues according to Steps 2 and 3 in Example 1 and Examples 16 - 19 using the same batch of Enterococcus faecium seed liquid and yeast seed liquid, and measure the viable yeast count in the fermented medicinal residues obtained by fermentation.

[0080] The results are as Figure 4 shown. When the fermentation days are 2, 3, and 4, there is no statistical difference in the viable yeast count. Among them, when the fermentation days are 3, the viable yeast count is the highest. When the fermentation days increase to 5, the viable yeast count decreases. When the fermentation days are only 1, the viable yeast count is the lowest.

[0081] Test Example 7 Prepare Enterococcus faecium seed liquid and yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2), and prepare fermented medicinal residues according to Steps 2 and 3 in Example 1 and Examples 20 - 24 respectively using the same batch of Enterococcus faecium seed liquid and yeast seed liquid, and measure the viable yeast count in the fermented medicinal residues obtained by fermentation.

[0082] The results are as Figure 5 shown. As the proportion of yeast in the mixed seed liquid increases, the viable yeast count in the fermented medicinal residues shows a trend of first increasing and then decreasing. Under the conditions of a mixing ratio of 1:1 - 1:3, the viable yeast count in the fermented medicinal residues is relatively high and there is no statistical difference. When the mixing ratio is 1:1, the viable yeast count is slightly higher than that of other groups. When the mixing ratio is 1:4, the viable yeast count decreases.

[0083] Test Example 8 Prepare Enterococcus faecium seed liquid and yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2), and prepare fermented medicinal residues according to Steps 2 and 3 in Example 1 and Examples 25 - 28 respectively using the same batch of Enterococcus faecium seed liquid and yeast seed liquid, and measure the viable yeast count in the fermented medicinal residues obtained by fermentation.

[0084] The results are as Figure 6 shown. The viable yeast count in the fermented medicinal residues first increases and then decreases as the inoculation amount of the mixed seed liquid increases. When the inoculation amount of the mixed seed liquid is 8%, the viable yeast count in the fermented medicinal residues is the largest. When the inoculation amount of the mixed seed liquid continues to increase to 10%, the viable yeast count decreases.

[0085] Test Example 9 Prepare Enterococcus faecium seed liquid and yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2), and use the same batch of Enterococcus faecium seed liquid and yeast seed liquid to prepare fermented medicinal residues according to Steps 2 and 3 in Example 1 and Examples 29 to 33 respectively, and measure the viable yeast count of the fermented medicinal residues obtained by fermentation.

[0086] The results are as Figure 7 shown. The influence of the fermentation temperature within 25°C to 32°C on the viable yeast count is not significant, with small differences. At 30°C, the viable yeast count is the highest. When the fermentation temperature continues to rise to 35 - 38°C, the viable cell count decreases significantly.

[0087] Test Example 10 Prepare Enterococcus faecium seed liquid and yeast seed liquid according to the method of Example 1 (Enterococcus faecium OD 560 = 0.7, yeast OD 600 = 1.2), and use the same batch of Enterococcus faecium seed liquid and yeast seed liquid to prepare fermented medicinal residues according to Steps 2 and 3 in Example 37, and measure the viable yeast count of the fermented medicinal residues on the 3rd, 6th, 9th, 12th, and 15th days after inoculating the mixed seed liquid and starting fermentation.

[0088] The results are as Figure 8 shown. After inoculating the mixed seed liquid, under the condition of fermentation for 6 - 9 days, the viable yeast count is relatively high and there is no significant difference. Among them, when the fermentation days are 6 days, the viable yeast count is the highest. As the fermentation time extends, when fermenting to 12 - 15 days, the viable yeast count decreases.

[0089] Example 41 This example provides a kind of fermented medicinal residue.

[0090] The preparation method is the same as that of Example 1.

[0091] Take the solid fermentation medium at 0 day after inoculation and at the end of fermentation, that is, the 9th day, to measure the viable yeast count, crude protein content, acid-soluble protein content, free amino acid content, total phosphorus, total potassium content, total phenol content, DPPH scavenging rate, and hydroxyl radical scavenging rate, and evaluate the nutritional value of the fermented medicinal residue.

[0092] The results are shown in Table 3. The viable cell count after fermentation reaches 1.28×10 8CFU / g, the crude protein content, acid-soluble protein content, and free amino acid content were significantly increased after fermentation, with increases of 25.45%, 57.03%, and 22.57% respectively compared to before fermentation. The total phosphorus and total potassium contents both decreased slightly. In terms of in vitro antioxidant activity, the total phenols, DPPH scavenging rate, and hydroxyl radical scavenging rate were significantly increased after fermentation, which helped to scavenge free radicals, inhibit pathogens, and thus promote the growth of livestock.

[0093] Table 3 Analysis of nutritional components before and after fermentation

[0094] Example 42 This example investigated the effects of the fermented medicinal residues prepared in Example 1 on the growth performance and diarrhea rate of transport-stressed meat sheep.

[0095] Eighteen healthy meat sheep (male, 3 months old) with similar body weights and body conditions were randomly divided into two groups: a control group (without adding fermented medicinal residues) and an experimental group (additionally adding 10% fermented medicinal residues). The body weight on the day before transportation was recorded as the initial weight, and the body weight 15 days after transportation was recorded as the end weight. All experimental sheep were transported on a 5-hour road journey on the day of transportation. The transportation conditions, breeding conditions, and feeding management were carried out according to the daily management regulations. The control group was fed a basal diet before and after transportation; the experimental group was fed a basal diet supplemented with 10% fermented medicinal residues from 3 days before transportation to 7 days after transportation.

[0096] The results are shown in Table 4. Feeding meat sheep with fermented medicinal residues added to the basal diet can alleviate the negative impacts of transport stress on the slow weight gain and reduced feed intake of meat sheep, reduce the damage caused by transportation to meat sheep, lower the diarrhea rate of meat sheep after transport stress, and improve the breeding efficiency.

[0097] Table 4 Effects of adding fermented medicinal residues on the growth performance of transport-stressed meat sheep

[0098] Example 43 This example investigated the effects of the fermented medicinal residues prepared in Example 1 on the growth performance of healthy meat sheep.

[0099] Forty-eight healthy meat sheep (male, 3 months old) with similar body weights and body conditions were randomly divided into 4 groups, with 12 replicates in each group and 1 sheep in each replicate. Each group was fed a basal diet supplemented with 0%, 3%, 5%, and 10% fermented medicinal residues respectively. The preliminary trial period was 1 week, and the formal trial period was 30 days. Before the start of the trial, the breeding house was disinfected, and deworming was carried out uniformly. Feeding was carried out twice a day at 7:30 and 17:30 respectively, and drinking water was provided ad libitum. The sheep house, water trough, and feeding trough were cleaned and disinfected every week. The immunization and deworming procedures for the experimental sheep were carried out according to the daily management regulations.

[0100] Before the morning feeding on the 1st day and the last day of the formal trial, the fasting body weights of each group of meat sheep were weighed as the initial weight and the ending weight. During the formal trial, the feed intake of each meat sheep was accurately recorded, and the average daily gain, average daily feed intake, and feed-to-gain ratio were calculated. Average daily feed intake = total feed intake / number of trial days; average daily gain = total weight gain / number of trial days; feed-to-gain ratio = average daily feed intake / average daily gain.

[0101] As shown in Table 5, when 0%, 3%, 5%, and 10% of fermented medicinal residues were added to the basal diet to feed the meat sheep, the average daily gain and average feed intake of the meat sheep were significantly increased, the feed-to-gain ratio was reduced, and the growth performance of the meat sheep was improved.

[0102] Table 5 Effects of adding fermented medicinal residues on the growth performance of meat sheep

[0103] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, and improvements made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A preparation method of fermented medicinal residues, characterized in that, Specifically, it includes the following steps: S1. Activate lactic acid bacteria and yeast, and culture them until the logarithmic growth phase to obtain lactic acid bacteria and yeast seed solutions respectively; dilute the yeast fermentation residue liquid by 5 - 8 times to obtain a diluted yeast fermentation residue liquid; mix the diluted yeast fermentation residue liquid, radix isatidis residue, and corncob powder according to the mass ratio of (1.5 - 9):2:(0.8 - 1.2), add urea accounting for 0.5% - 3.0% of the mass of the obtained mixture, and stir evenly to obtain a solid fermentation medium; S2. Inoculate the solid fermentation medium with the lactic acid bacteria seed solution at an inoculation amount of 1% - 5%, stir evenly and ferment at 25 - 38°C for 1 - 5 days, then inoculate a mixed seed solution with a volume ratio of (1 - 3):(1 - 4) of the lactic acid bacteria seed solution and the yeast seed solution at an inoculation amount of 2% - 10%, stir evenly and ferment at 25 - 38°C for 3 - 15 days.

2. The preparation method according to claim 1, characterized in that, The lactic acid bacteria in S1 are Enterococcus faecium; and / or The yeast in S1 is Saccharomyces cerevisiae; and / or The lactic acid bacteria and yeast in S1 are selected from strains that can grow well in a medium containing yeast fermentation residue liquid; and / or The mass ratio of the diluted yeast fermentation residue liquid, radix isatidis residue, and corncob powder in S1 is 6:2:1; and / or The addition amount of urea in S1 is 1.0% - 2.0%; and / or The inoculation amount of the lactic acid bacteria seed solution in S2 is 1% - 3%; and / or The fermentation time after inoculating the lactic acid bacteria seed solution in S2 is 2 - 4 days; and / or The volume ratio of the lactic acid bacteria seed solution to the yeast seed solution in the mixed seed solution in S2 is 1:(1 - 3); and / or The inoculation amount of the mixed seed solution in S2 is 6% - 8%; and / or The fermentation time after inoculating the mixed seed solution in S2 is 6 - 9 days; and / or The fermentation temperature in S2 is 25 - 32°C.

3. The preparation method according to claim 1 or 2, characterized in that, The screening method for the lactic acid bacteria and yeast is: Activate the lactic acid bacteria and yeast to be screened, inoculate them into the yeast fermentation residue liquid medium at an inoculation amount of 3% for culture, select the strain with a high viable cell count for subculture, continue to culture the subcultured strain in the yeast fermentation residue liquid medium, repeat the operation until 10 - 15 generations of subculture, and then select the strain with the highest viable cell count, which is the lactic acid bacteria and yeast in S1; the composition of the yeast fermentation residue liquid medium is: yeast fermentation residue liquid and water with a volume ratio of 1:4, and 10 g / L urea; and / or The addition amount of urea in S1 is 1.5% - 2.0%; and / or The inoculation amount of the lactic acid bacteria seed solution in S2 is 2%; and / or The fermentation time after inoculating the lactic acid bacteria seed solution in S2 is 3 days; and / or The volume ratio of the lactic acid bacteria seed solution to the yeast seed solution in the mixed seed solution in S2 is 1:1; and / or The inoculation amount of the mixed seed solution in S2 is 8%; and / or The fermentation time after inoculating the mixed seed solution in S2 is 6 days; and / or The fermentation temperature in S2 is 30°C.

4. The fermented residue prepared by the preparation method according to any one of claims 1 - 3.

5. Use of the fermented medicinal residues according to claim 4 as a feed additive.

6. The application according to claim 5, wherein The feed additive is a sheep feed additive.

7. The application according to claim 6, wherein The feed additive is a meat sheep feed additive.

8. A method for alleviating transportation stress and / or reducing diarrhea rate in meat sheep, characterized in that, Add 5% - 10% wt of the fermented medicinal residues according to claim 4 to the meat sheep diet.

9. A method for improving the growth performance of meat sheep, characterized in that, Add 3% - 10% wt of the fermented medicinal residues according to claim 4 to the meat sheep diet.

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