A feed additive for preventing diarrhea of piglets and its preparation method and application

Abstract

The present application provides a preparation method of a feed additive for preventing piglet diarrhea, comprising the following steps: preparing a fermentation base; fermenting the fermentation base to obtain a pre-fermentation material; preparing a trace element fermentation liquid, which is added to the pre-fermentation material, and a mixed bacteria liquid and an enzyme are added for culture to obtain the feed additive. The feed additive prepared by the present application uses traditional Chinese medicine as raw material, and through the synergistic fermentation of traditional Chinese medicine, bacteria, trace elements and enzymes, a new formula is prepared, and the feed additive for preventing piglet diarrhea has innovation and huge market space.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pig feeding feed additive, and particularly relates to a feed additive for preventing diarrhea of piglets and a preparation method and application thereof. BACKGROUND

[0002] China is a big country of pig raising, and pork consumption is huge. Chinese medicine is a competitive resource in China, and developing new formula is an important direction of application of Chinese medicine in the breeding industry. Chinese medicine has the advantages of safety, greenness, high efficiency, non-toxicity and the like, and Chinese medicine is a characteristic and advantageous resource in China. Chinese medicine resources are various in types and good in quality.

[0003] Scientific use of trace elements is a scientific way to improve breeding benefits. Trace elements are essential components for livestock and poultry, and have multiple functions. Trace elements are not only essential elements for the activity of various enzymes in the animal body, but also essential elements for growth metabolism, development metabolism, immune metabolism and disease resistance metabolism of animals. There are various probiotics, and the role of probiotics in livestock and poultry breeding is becoming more and more significant. Probiotics have the effect of Chinese medicine in maintaining animal intestinal health, improving disease resistance and immune capacity, and improving the efficacy of Chinese medicine.

[0004] Scientific use of enzymes can achieve the purpose of improving breeding benefits from multiple aspects. Enzymes can not only promote digestion and absorption, but also promote the release of effective components of Chinese medicine, the exertion of health or nutritional effects of trace elements, and the like. The production and survival status of piglets are directly related to the level of breeding benefits.

[0005] There are various diseases of piglets, and the incidence of diarrhea is high, which is harmful to piglet breeding. Piglets are caused by nutritional changes such as weaning, and factors such as microbial diseases and environmental stimulation, which will cause diarrhea of piglets in the rearing stage. Diarrhea reduces the growth benefit of piglets, and directly or indirectly increases the breeding cost.

[0006] Chinese patent CN102919607B provides a Chinese herbal medicine compound pig feed additive for preventing and treating pig diarrhea and improving pork quality, which is composed of the following components: wormwood, hawthorn leaf, purslane, poria cocos, dried tangerine or orange peel, sophora flavescens, polyporus, copper sulfate, manganese sulfate, sodium bicarbonate, sodium selenate, lysine, methionine, salt and zeolite powder. The Chinese herbal medicine components are treated by simple extraction.

[0007] The treatment of Chinese medicine components in the above-mentioned prior art only simply utilizes the efficacy of Chinese medicine, and the effect of preventing piglet diarrhea needs to be further improved. Therefore, through scientific formula, by using Chinese medicine as a raw material for fermentation, by using the compound fermentation of bacteria, Chinese medicine and enzymes, a new formula is produced to produce a feed additive for preventing diarrhea of piglets, which has innovation and huge market space. SUMMARY

[0008] Based on the technical problems existing in the background art, the present application provides a feed additive for preventing diarrhea of piglets and a preparation method and application thereof, adopts traditional Chinese medicine as a fermentation raw material, utilizes multiple strains and enzymes for compound fermentation, produces the green feed additive for preventing diarrhea of piglets through a new traditional Chinese medicine prescription and a new process, and improves the green breeding benefit of piglets.

[0009] In order to achieve the above-mentioned purpose, the technical scheme of the present application is as follows:

[0010] A preparation method of a feed additive for preventing diarrhea of piglets, comprising the following steps:

[0011] (1) Preparation of fermentation base material: mix mulberry leaves, radix puerariae, gastrodia tuber, flaxseed, medlar, jujube, tuckahoe, konjac powder and Chinese yam, crush and sieve to obtain traditional Chinese medicine mixed powder; add cellulase, hemicellulase, pectinase and sterilized water to the traditional Chinese medicine mixed powder, and incubate at 45-55 DEG C for 10-14 h to obtain enzyme hydrolysis primary wet powder;

[0012] Add proteose peptone and glucose to the enzyme hydrolysis primary wet powder to obtain fermentation base material;

[0013] (2) Preparation of pre-fermentation material: inoculate the first mixed bacteria liquid into the fermentation base material, and ferment at 25-30 DEG C for 45-50 h to obtain pre-fermentation material;

[0014] (3) Preparation of trace element fermentation material liquid: prepare trace element fermentation medium, inoculate the second mixed bacteria liquid into the trace element fermentation medium, and then add manganese lactate and copper methionine, mix uniformly, and then perform primary culture; after the culture is completed, the trace element primary fermentation material liquid is obtained;

[0015] Add keratinase, glucose and proteose peptone to the obtained trace element primary fermentation material liquid, mix uniformly, and then perform secondary culture; after the culture is completed, the trace element fermentation material liquid is obtained;

[0016] (4) Preparation of feed additive: add the trace element fermentation material liquid, lactococcus lactis bacteria liquid, saccharomyces cerevisiae bacteria liquid, cellulase, hemicellulase, pectinase and galactosidase to the pre-fermentation material obtained in step (2), and incubate at 25-30 DEG C for 70-75 h to obtain fermentation enzyme hydrolysis whole material;

[0017] Inoculate laccase into the fermentation enzyme hydrolysis whole material, mix uniformly, incubate at 40-50 DEG C for 34-38 h, then control the temperature to 72-78 DEG C for 2-4 h, and obtain the feed additive.

[0018] Preferably, in step (1), the mass ratio of mulberry leaves, radix puerariae, rhizoma gastrodiae, linum usitatissimum, medlar, jujube, tuckahoe, konjac powder, dioscorea is 4-8:0.1-2:0.5-4:0.1-2:0.01-1:7-11:0.1-2:0.5-4:0.5-4.

[0019] Preferably, in step (1), the amount of cellulase, hemicellulase, pectinase and sterilized water is 5%, 2%, 1% and 58% of the mass of the traditional Chinese medicine mixed powder, respectively.

[0020] Preferably, in step (1), the amount of proteose peptone and glucose is 2% and 4% of the mass of the primary wet powder after enzymolysis, respectively.

[0021] Preferably, in step (2), the composition of the first mixed bacteria solution is: Saccharomyces cerevisiae bacteria solution, Bacillus subtilis bacteria solution, Bacillus licheniformis bacteria solution, Bacillus coagulans bacteria solution, Lactobacillus delbrueckii bacteria solution, Lactobacillus plantarum bacteria solution, Clostridium butyricum bacteria solution and Pediococcus acidilactici bacteria solution, and the amount is 1%, 1%, 1%, 2%, 1%, 1%, 1% and 3% of the mass of the fermentation base material, respectively.

[0022] Preferably, in step (3), the composition of the second mixed bacteria solution is: Saccharomyces cerevisiae bacteria solution, Bacillus coagulans bacteria solution, Clostridium butyricum bacteria solution and Pediococcus acidilactici bacteria solution, and the amount is 1%, 2%, 1% and 3% of the mass of the trace element fermentation medium, respectively.

[0023] The amount of manganese lactate and copper methionine is 0.2% and 0.15% of the mass of the trace element fermentation medium, respectively.

[0024] The initial culture temperature is 29-31°C, and the culture time is 24-32h.

[0025] Preferably, in step (3), the amount of keratinase, glucose and proteose peptone is 0.1%, 1% and 0.2% of the mass of the trace element primary fermentation broth, respectively.

[0026] The secondary culture temperature is 29-31°C, and the culture time is 24-32h.

[0027] Preferably, in step (4), the amount of trace element fermentation broth is 10% of the mass of the pre-fermentation material.

[0028] The amount of Pediococcus acidilactici bacteria solution and Saccharomyces cerevisiae bacteria solution is 1% and 2% of the mass of the pre-fermentation material, respectively.

[0029] The amount of cellulase, hemicellulase, pectinase and galactoside enzyme is 1%, 2%, 0.5% and 1% of the mass of the pre-fermentation material, respectively.

[0030] The amount of laccase is 3% of the mass of the fermentation enzyme hydrolysis whole material.

[0031] In a second aspect, the present application also claims a feed additive prepared by the above method.

[0032] In a third aspect, the present application also claims the use of the above feed additive in preventing diarrhea of piglets.

[0033] Compared with the prior art, the present application has the following beneficial effects:

[0034] The feed additive provided by the present application is prepared from nine Chinese herbal medicines, i.e., mulberry leaves, radix puerariae, rhizoma gastrodiae, linseed, medlar, red dates, poria cocos, konjac powder and Chinese yam, which can synergistically exert the maximum effect of preventing diarrhea of piglets. In addition, the synergistic effect of Chinese herbal medicines, trace elements, probiotics and enzymes in the preparation process can achieve a synergistic effect, thereby enhancing the resistance, promoting the absorption of nutrients and improving the intestinal flora, so that the prepared feed additive can exert the best pharmacodynamic effect of preventing diarrhea of piglets. DETAILED DESCRIPTION

[0035] The technical solutions in the embodiments of the present application will be clearly and completely described in combination with the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0036] Unless otherwise specified, all goods or reagents in the present application are purchased through market channels.

[0037] The Saccharomyces cerevisiae, Clostridium butyricum, Lactobacillus plantarum and Bacillus subtilis are purchased from Qingdao Blue Bio-technology Co., Ltd.

[0038] The Bacillus coagulans is purchased from Shandong Chenzhan Bio-technology Co., Ltd.

[0039] The Lactobacillus delbrueckii is purchased from Shanghai Guyan Industry Co., Ltd.

[0040] The Pediococcus acidilactici is purchased from Weipei (Guangdong) Bio-technology Co., Ltd.

[0041] The Bacillus licheniformis is purchased from Hunan Longkai Pharmaceutical Co., Ltd.

[0042] The cellulase is purchased from Ningxia Xiasen Industry Group Co., Ltd., 11000 U / g.

[0043] The hemicellulase is purchased from Ningxia Xiasen Industry Group Co., Ltd., 50000 U / g.

[0044] The pectinase is purchased from Ningxia Xiasen Industry Group Co., Ltd., H type / viscosity reduction, 25000 U / g

[0045] Galactosidase was purchased from Sheng Group, 500u / g;

[0046] Laccase was purchased from Sheng Group, 1000U / g;

[0047] Mulberry leaves were purchased from Yunnan Nuolin Biological Technology Co., Ltd.;

[0048] Kudzu was purchased from Bozhou Tiaoyangtang Pharmaceutical Co., Ltd.;

[0049] Gastrodia was purchased from Huamei Medicinal Materials;

[0050] Flax seeds were purchased from Anguo Jianhao Medicinal Materials;

[0051] Medlar was purchased from Ningxia Qicai Hometown Specialty Development Co., Ltd.;

[0052] Red dates were purchased from Cangzhou Babaochi Food Co., Ltd.;

[0053] Poria cocos was purchased from Bozhou Anfutang Pharmaceutical Co., Ltd.;

[0054] Konjac powder was purchased from Hunan Century Huaxing Biological Engineering Co., Ltd.;

[0055] Chinese yam was purchased from Bozhou Baohuatao Biological Technology Co., Ltd.;

[0056] Proteose peptone was purchased from Jinan Lanjue Business and Trade Co., Ltd.;

[0057] Glucose was purchased from Shandong Xujie Chemical Industry Co., Ltd.;

[0058] Yeast extract was purchased from Jinan Xinsenyuan Chemical Industry Co., Ltd.;

[0059] Manganese lactate was purchased from Henan Qiaoshou Food Additive Co., Ltd.;

[0060] Copper methionine was purchased from Hebei Hongtao Biological Engineering Co., Ltd.

[0061] In the embodiments and comparative examples of the present application, the activation preparation of the bacterial liquid is as follows:

[0062] The six rings of bacteria were scraped with an inoculation ring (ring diameter 0.3mm) from the slant preserved Saccharomyces cerevisiae, inoculated into 300mL of culture medium, and loaded into a 500mL flask, which was cultured at 33℃ with a rotation speed of 180r / m for 18h to obtain the Saccharomyces cerevisiae bacterial liquid.

[0063] The six rings of bacteria were scraped with an inoculation ring (ring diameter 0.3mm) from the slant preserved Bacillus subtilis, inoculated into 300mL of culture medium, and loaded into a 500mL flask, which was cultured at 30℃ with a rotation speed of 180r / m for 18h to obtain the Bacillus subtilis bacterial liquid.

[0064] The bacillus licheniformis preserved on the slope is scraped with an inoculation ring (ring diameter 0.3 mm) to obtain six ring bacteria, which are inoculated into 300 mL of culture medium and loaded into a 500 mL flask, and then cultured at 30°C with a rotation speed of 180 r / m for 18 h to obtain bacillus licheniformis liquid.

[0065] The lactobacillus plantarum strain preserved on the slope is scraped with an inoculation ring (ring diameter 0.3 mm) to obtain six ring bacteria, which are inoculated into 300 mL of culture medium and loaded into a 500 mL flask, and then cultured at 30°C for 28 h to obtain lactobacillus plantarum liquid.

[0066] The clostridium butyricum strain preserved on the slope is scraped with an inoculation ring (ring diameter 0.3 mm) to obtain six ring bacteria, which are inoculated into 300 mL of culture medium and loaded into a 500 mL flask, and then cultured at 30°C for 28 h to obtain clostridium butyricum liquid.

[0067] The bacillus coagulans preserved on the slope is scraped with an inoculation ring (ring diameter 0.3 mm) to obtain six ring bacteria, which are inoculated into 300 mL of culture medium and loaded into a 500 mL flask, and then cultured at 30°C for 28 h to obtain bacillus coagulans liquid.

[0068] The lactobacillus delbrueckii preserved on the slope is scraped with an inoculation ring (ring diameter 0.3 mm) to obtain six ring bacteria, which are inoculated into 300 mL of culture medium and loaded into a 500 mL flask, and then cultured at 30°C for 28 h to obtain lactobacillus delbrueckii liquid.

[0069] The pediococcus acidilactici preserved on the slope is scraped with an inoculation ring (ring diameter 0.3 mm) to obtain six ring bacteria, which are inoculated into 300 mL of culture medium and loaded into a 500 mL flask, and then cultured at 30°C for 28 h to obtain pediococcus acidilactici liquid.

[0070] In the embodiments and comparative examples of the present application, the preparation steps of the culture medium include: 1% (w / w) of yeast extract, 1% (w / w) of protein peptone and 4% (w / w) of glucose; after preparation, sterilization is performed at 115°C for 30 min, and then the medium is cooled to room temperature.

[0071] The preparation steps of the trace element fermentation medium include: 2% (w / w) of yeast extract, 1% (w / w) of protein peptone and 6% (w / w) of glucose; after preparation, sterilization is performed at 115°C for 30 min, and then the medium is cooled to room temperature.

[0072] Example 1

[0073] A preparation method of a feed additive for preventing piglet diarrhea includes the following steps:

[0074] (1) Preparation of fermentation base material: mulberry leaves, radix puerariae, gastrodia, flaxseed, medlar, jujube, tuckahoe, konjac powder, yam were mixed according to the mass ratio of 6:1:2:1:0.5:9:1:2:2, crushed through a 100 mesh sieve, and a traditional Chinese medicine mixed powder was obtained; cellulase, hemicellulase, pectinase and sterilized water were added into the traditional Chinese medicine mixed powder respectively, and the amount of cellulase, hemicellulase, pectinase and sterilized water was 5%, 2%, 1% and 58% of the mass of the traditional Chinese medicine mixed powder respectively, and the mixture was incubated at 50℃ for 12h, and an enzymolysis primary wet powder was obtained;

[0075] Proteose peptone and glucose were added into the enzymolysis primary wet powder, and a fermentation base material was obtained; the amount of proteose peptone and glucose was 2% and 4% of the mass of the enzymolysis primary wet powder respectively;

[0076] (2) Preparation of pre-fermentation material: the first mixed bacteria liquid (Saccharomyces cerevisiae bacteria liquid, Bacillus subtilis bacteria liquid, Bacillus licheniformis bacteria liquid, Bacillus coagulans bacteria liquid, Lactobacillus delbrueckii bacteria liquid, Lactobacillus plantarum bacteria liquid, Clostridium butyricum bacteria liquid and Pediococcus acidilactici bacteria liquid) was inoculated into the fermentation base material, and the amount of the first mixed bacteria liquid was 1%, 1%, 1%, 2%, 1%, 1%, 1% and 3% of the mass of the fermentation base material respectively, and the mixture was fermented at 28℃ for 48h, and a pre-fermentation material was obtained;

[0077] (3) Preparation of trace element fermentation material liquid: the second mixed bacteria liquid (Saccharomyces cerevisiae bacteria liquid, Bacillus coagulans bacteria liquid, Clostridium butyricum bacteria liquid and Pediococcus acidilactici bacteria liquid) was inoculated into the prepared trace element fermentation medium, and the amount of the second mixed bacteria liquid was 1%, 2%, 1% and 3% of the mass of the trace element fermentation medium respectively, and then manganese lactate and copper methionine with the amount of 0.2% and 0.15% of the mass of the trace element fermentation medium were added, and the mixture was uniformly mixed and then cultured at 30℃ for 30h, and a trace element primary fermentation material liquid was obtained;

[0078] Keratinase, glucose and proteose peptone with the amount of 0.1%, 1% and 0.2% of the mass of the trace element primary fermentation material liquid were added into the obtained trace element primary fermentation material liquid respectively, and the mixture was uniformly mixed and then cultured again, and the mixture was cultured at 30℃ for 30h, and a trace element fermentation material liquid was obtained;

[0079] (4) Preparation of feed additive: the trace element fermentation material liquid with the amount of 10%, Pediococcus acidilactici bacteria liquid with the amount of 1%, Saccharomyces cerevisiae bacteria liquid with the amount of 2%, cellulase with the amount of 1%, hemicellulase with the amount of 2%, pectinase with the amount of 0.5% and galactoside enzyme with the amount of 1% of the mass of the pre-fermentation material obtained in step (2) were added into the pre-fermentation material, and the mixture was incubated at 28℃ for 72h, and a fermentation enzyme hydrolysis whole material was obtained;

[0080] Laccase with the amount of 3% of the mass of the fermentation enzyme hydrolysis whole material was inoculated into the fermentation enzyme hydrolysis whole material, and the mixture was uniformly mixed, incubated at 45℃ for 36h, and then incubated at 75℃ for 3h, and a feed additive was obtained.

[0081] Example 2

[0082] A method for preparing a feed additive for preventing diarrhea of piglets, comprising the following steps:

[0083] (1) Preparation of fermentation base material: mulberry leaves, radix puerariae, gastrodia, flaxseed, medlar, jujube, tuckahoe, konjac powder, yam are mixed in a mass ratio of 4.5:0.1:0.5:0.1:0.03:7:0.3:2.5:1.5, crushed to 100 mesh, and a traditional Chinese medicine mixed powder is obtained; cellulase, hemicellulase, pectinase and sterilized water are added to the traditional Chinese medicine mixed powder respectively, with the amount of cellulase being 5% of the mass of the traditional Chinese medicine mixed powder, the amount of hemicellulase being 2%, the amount of pectinase being 1%, and the amount of sterilized water being 58%, and the mixture is incubated at 50°C for 12 hours to obtain an enzymolysis primary wet powder;

[0084] Proteose peptone and glucose are added to the enzymolysis primary wet powder to obtain a fermentation base material; the amount of proteose peptone and glucose is 2% and 4% of the mass of the enzymolysis primary wet powder respectively;

[0085] (2) Preparation of pre-fermentation material: the fermentation base material is inoculated with a first mixed bacteria solution (Saccharomyces cerevisiae bacteria solution, Bacillus subtilis bacteria solution, Bacillus licheniformis bacteria solution, Bacillus coagulans bacteria solution, Lactobacillus delbrueckii bacteria solution, Lactobacillus plantarum bacteria solution, Clostridium butyricum bacteria solution and Pediococcus acidilactici bacteria solution, with the amount being 1%, 1%, 1%, 2%, 1%, 1%, 1% and 3% of the mass of the fermentation base material respectively), and the mixture is fermented at 28°C for 48 hours to obtain a pre-fermentation material;

[0086] (3) Preparation of trace element fermentation material solution: a second mixed bacteria solution (Saccharomyces cerevisiae bacteria solution, Bacillus coagulans bacteria solution, Clostridium butyricum bacteria solution and Pediococcus acidilactici bacteria solution, with the amount being 1%, 2%, 1% and 3% of the mass of the trace element fermentation medium respectively) is inoculated into the prepared trace element fermentation medium, and then manganese lactate and copper methionine with the amount of 0.2% and 0.15% of the mass of the trace element fermentation medium are added, and the mixture is uniformly mixed and then cultured at 30°C for 30 hours to obtain a trace element primary fermentation material solution;

[0087] Keratinase, glucose and proteose peptone with the amount of 0.1%, 1% and 0.2% of the mass of the trace element primary fermentation material solution are added into the obtained trace element primary fermentation material solution, and the mixture is uniformly mixed and then cultured again at 30°C for 30 hours to obtain a trace element fermentation material solution;

[0088] (4) Preparation of feed additive: the pre-fermentation material obtained in step (2) is inoculated with a trace element fermentation material solution, Pediococcus acidilactici bacteria solution, Saccharomyces cerevisiae bacteria solution, cellulase, hemicellulase, pectinase and galactosidase with the amount of 10%, 1%, 2%, 1%, 2%, 0.5% and 1% of the mass of the pre-fermentation material respectively, and the mixture is incubated at 28°C for 72 hours to obtain a fermentation enzyme hydrolysis whole material;

[0089] The laccase with a mass of 3% is inoculated into the fermentation enzymolysis whole material, mixed uniformly, and incubated at 45℃ for 36h, and then incubated at 75℃ for 3h, to obtain the feed additive.

[0090] Example 3

[0091] A preparation method of a feed additive for preventing piglet diarrhea, comprising the following steps:

[0092] (1) Preparation of fermentation base material: mix mulberry leaves, radix puerariae, gastrodia, flaxseed, medlar, jujube, tuckahoe, konjac powder, and Chinese yam according to a mass ratio of 8:2:4:2:1:11:2:4:4, crush and pass through an 80-mesh sieve to obtain a traditional Chinese medicine mixed powder; add cellulase, hemicellulase, pectinase, and sterilized water to the traditional Chinese medicine mixed powder, with the amounts of the cellulase, hemicellulase, pectinase, and sterilized water being 5%, 2%, 1%, and 58% of the mass of the traditional Chinese medicine mixed powder respectively, and incubate at 50℃ for 12h to obtain an enzymolysis primary wet powder;

[0093] Add protein peptone and glucose to the enzymolysis primary wet powder to obtain a fermentation base material; the amounts of the protein peptone and glucose are 2% and 4% of the mass of the enzymolysis primary wet powder respectively;

[0094] (2) Preparation of pre-fermentation material: inoculate the fermentation base material with a first mixed bacteria solution (Saccharomyces cerevisiae bacteria solution, Bacillus subtilis bacteria solution, Bacillus licheniformis bacteria solution, Bacillus coagulans bacteria solution, Lactobacillus delbrueckii bacteria solution, Lactobacillus plantarum bacteria solution, Clostridium butyricum bacteria solution, and Pediococcus acidilactici bacteria solution, with the amounts being 1%, 1%, 1%, 2%, 1%, 1%, 1%, and 3% of the mass of the fermentation base material respectively), and ferment at 28℃ for 48h to obtain a pre-fermentation material;

[0095] (3) Preparation of trace element fermentation material solution: inoculate the prepared trace element fermentation medium with a second mixed bacteria solution (Saccharomyces cerevisiae bacteria solution, Bacillus coagulans bacteria solution, Clostridium butyricum bacteria solution, and Pediococcus acidilactici bacteria solution, with the amounts being 1%, 2%, 1%, and 3% of the mass of the trace element fermentation medium respectively), and then add manganese lactate and copper methionine with the amounts being 0.2% and 0.15% of the mass of the trace element fermentation medium respectively, mix uniformly, and then culture at 30℃ for 30h to obtain a trace element primary fermentation material solution;

[0096] Add keratinase, glucose, and protein peptone to the obtained trace element primary fermentation material solution with the amounts being 0.1%, 1%, and 0.2% of the mass of the trace element primary fermentation material solution respectively, mix uniformly, and then culture again at 30℃ for 30h to obtain the trace element fermentation material solution;

[0097] (4) Preparation of the feed additive: 10% of the trace element fermentation material, 1% of the Pediococcus acidilactici bacterial liquid, 2% of the Saccharomyces cerevisiae bacterial liquid, 1% of the cellulase, 2% of the hemicellulase, 0.5% of the pectinase and 1% of the galactoside enzyme are added into the pre-fermentation material obtained in step (2), and then the mixture is incubated at 28°C for 72 hours to obtain the fermentation and enzymolysis whole material;

[0098] Then, 3% of the laccase is inoculated into the fermentation and enzymolysis whole material, and the mixture is uniformly mixed and incubated at 45°C for 36 hours, and then the temperature is controlled at 75°C for 3 hours to obtain the feed additive.

[0099] Comparative Example 1

[0100] A preparation method of a feed additive for preventing piglet diarrhea, comprising the following steps:

[0101] (1) Preparation of the fermentation base material: the mulberry leaves, the radix puerariae, the Gastrodia elata Blume, the flaxseed, the medlar, the jujube, the Poria cocos, the konjac powder and the Chinese yam are mixed according to the mass ratio of 4.5:0.1:0.5:0.1:0.03:7:0.3:2.5:1.5, and then the mixture is crushed and passed through an 80-mesh sieve to obtain a traditional Chinese medicine mixed powder; the traditional Chinese medicine mixed powder is added with 5% of the cellulase, 2% of the hemicellulase, 1% of the pectinase and 58% of the sterilized water, and then the mixture is incubated at 50°C for 12 hours to obtain an enzymolysis primary wet powder;

[0102] Then, the protein peptone and the glucose are added into the enzymolysis primary wet powder to obtain the fermentation base material; the added amounts of the protein peptone and the glucose are 2% and 4% of the mass of the enzymolysis primary wet powder, respectively;

[0103] (2) Preparation of the pre-fermentation material: the fermentation base material is inoculated with the first mixed bacterial liquid (the Saccharomyces cerevisiae bacterial liquid, the Bacillus subtilis bacterial liquid, the Bacillus licheniformis bacterial liquid, the Bacillus coagulans bacterial liquid, the Lactobacillus delbrueckii bacterial liquid, the Lactobacillus plantarum bacterial liquid, the Clostridium butyricum bacterial liquid and the Pediococcus acidilactici bacterial liquid, and the amounts of the bacterial liquids are 1%, 1%, 1%, 2%, 1%, 1%, 1% and 3% of the mass of the fermentation base material, respectively), and then the mixture is fermented at 28°C for 48 hours to obtain the pre-fermentation material;

[0104] (3) Preparation of the trace element fermentation material liquid: the second mixed bacterial liquid (the Saccharomyces cerevisiae bacterial liquid, the Bacillus coagulans bacterial liquid, the Clostridium butyricum bacterial liquid and the Pediococcus acidilactici bacterial liquid, and the amounts of the bacterial liquids are 1%, 2%, 1% and 3% of the mass of the trace element fermentation medium, respectively) is inoculated into the prepared trace element fermentation medium, and then the manganese lactate and the copper methionine in an amount of 0.2% and 0.15% of the mass of the trace element fermentation medium, respectively, are added, and the mixture is uniformly mixed and then cultured at 30°C for 30 hours to obtain the trace element primary fermentation material liquid;

[0105] To the obtained trace element primary fermentation broth, keratinase, glucose and proteose peptone were added respectively, the mass of the trace element primary fermentation broth was 0.1%, 1% and 0.2% respectively, and then the mixture was uniformly mixed and cultured again, and the trace element fermentation broth was obtained after 30℃ culture for 30h;

[0106] (4) Preparation of the feed additive: to the pre-fermentation broth obtained in step (2), the trace element fermentation broth, lactococcus lactis, saccharomyces cerevisiae, cellulase, hemicellulase, pectinase and galactoside enzyme were added respectively, the mass of the trace element fermentation broth was 10%, the mass of the lactococcus lactis was 1%, the mass of the saccharomyces cerevisiae was 2%, the mass of the cellulase was 1%, the mass of the hemicellulase was 2%, the mass of the pectinase was 0.5% and the mass of the galactoside enzyme was 1%, and then the mixture was incubated at 28℃ for 72h, and the feed additive was obtained.

[0107] Comparative Example 2

[0108] Compared with Example 1, other than not adding konjac powder, other steps were the same as Example 1.

[0109] Comparative Example 3

[0110] Compared with Example 1, other than not adding mulberry leaves, other steps were the same as Example 1.

[0111] Comparative Example 4

[0112] Compared with Example 1, other than not adding manganese lactate, other steps were the same as Example 1.

[0113] Comparative Example 5

[0114] Compared with Example 1, other than not adding pueraria, other steps were the same as Example 1.

[0115] Comparative Example 6

[0116] Compared with Example 1, other than not adding gastrodia elata, other steps were the same as Example 1.

[0117] The feed additives prepared in Example 1 and Comparative Examples 1-6 were tested to study the influence of the feed additives on diarrhea of piglets, and the specific test method was as follows:

[0118] The 28-day weaned, similar parity, genetically similar, similar weight Duodachang three-way crossbred pigs are used as the test pigs, 400 piglets are prepared, the preparation period is 10 days, and the formal test period is 30 days. The piglets are divided into 8 groups, 5 replicates in each group, and 10 piglets in each group. The piglets in each group are respectively provided with a numbered ring on the neck. The piglets are fed with a trough feed (Haoyunlai good baby piglet trough feed, Hunan Jinhaoyuan Biological Science and Technology Co., Ltd.) in the preparation period, and the control group is fed with a basic diet (corn 70.15%, soybean meal 22.0%, fish meal (Tianjin Dingsheng Feed Co., Ltd.) 5.0%, calcium hydrogen phosphate 1%, stone powder 0.55%, salt 0.3%, piglet premix (Anhui Yiyuan Farming and Stockbreeding Technology Co., Ltd.) 1%) in the formal test period, and the experimental group is fed with the basic diet + 2% of the additive (i.e., the feed additive prepared in Example 1 and Comparative Examples 1-6).

[0119] The test piglets are in the same pig house but different pig pens. The piglets are uniformly raised, and the ventilation, cooling, and sanitary management are the same. The piglets are allowed to freely eat and drink, the trough is cleaned every morning, and the remaining feed is weighed. The feed is thrown in the trough, and the remaining feed is slightly more than the feed. The pig house is cleaned and disinfected every 7 days. The piglets are observed every 30 minutes a day, and the diarrhea and the number of piglets with diarrhea are recorded. The piglets with diarrhea are not repeated.

[0120] The number of piglets that die in the formal test period is counted. The results are shown in Table 1.

[0121] Table 1

[0122] Group Total number of heads Number of heads of diarrhea Number of heads of death Control group 50 28 7 Example 1 50 5 3 Comparative Example 1 50 10 5 Comparative Example 2 50 5 4 Comparative Example 3 50 7 4 Comparative Example 4 50 6 4 Comparative Example 5 50 7 5 Comparative Example 6 50 8 4

[0123] It can be seen that the feed additive obtained by the combination of the traditional Chinese medicine composition and the fermentation process can effectively prevent the diarrhea of piglets and reduce the mortality rate.

[0124] The above detailed disclosure is only used as an example for those skilled in the art, and does not limit the present specification. Although it is not explicitly stated here, those skilled in the art can make various modifications, improvements and deviations to the present specification. Such modifications, improvements and deviations are suggested in the present specification, so such modifications, improvements and deviations still belong to the spirit and scope of the exemplary embodiments of the present specification.

[0125] It should be noted that if the description, definition and / or term used in the attached materials of the present specification are inconsistent or conflicting with the description, definition and / or term used in the present specification, the description, definition and / or term used in the present specification shall prevail.

[0126] Finally, it should be understood that the embodiments described herein are only given by way of example and that other modifications can occur to persons skilled in the art. Therefore, the scope of the present disclosure is not intended to be limited to the embodiments described herein but is only limited by the claims that follow.

Claims

1. A method for preparing a feed additive to prevent diarrhea in piglets, characterized in that: Includes the following steps: (1) Preparation of fermentation substrate: Mulberry leaves, kudzu root, gastrodia elata, flax seeds, wolfberry, red dates, poria cocos, konjac powder, and yam are mixed, pulverized and sieved to obtain mixed Chinese medicine powder; cellulase, hemicellulase, pectinase and sterilized water are added to the mixed Chinese medicine powder, and the mixture is kept at 45-55℃ for 10-14 hours to obtain primary wet powder of enzymatic hydrolysis; peptone and glucose are added to the primary wet powder of enzymatic hydrolysis to obtain fermentation substrate; (2) Preparation of pre-fermented material: Inoculate the first mixed bacterial solution into the fermentation substrate and ferment at 25-30℃ for 45-50h to obtain pre-fermented material; (3) Preparation of micronutrient fermentation broth: Prepare micronutrient fermentation medium, inoculate it with a second mixed bacterial solution, then add manganese lactate and copper methionine, mix well and carry out the first culture; after the culture is completed, the micronutrient primary fermentation broth is obtained; add keratinase, glucose and peptone to the obtained micronutrient primary fermentation broth, mix well and carry out the second culture; after the culture is completed, the micronutrient fermentation broth is obtained. (4) Preparation of feed additives: Add trace element fermentation liquid, Pediococcus lactis liquid, Saccharomyces cerevisiae liquid, cellulase, hemicellulase, pectinase and galactosidase to the pre-fermented material obtained in step (2), keep warm at 25-30℃ for 70-75h to obtain fermented enzymatic hydrolysate; add laccase to the fermented enzymatic hydrolysate, mix evenly, keep warm at 40-50℃ for 34-38h, and then keep warm at 72-78℃ for 2-4h to obtain feed additives; In step (1), the mass ratio of mulberry leaves, kudzu root, gastrodia elata, flax seeds, wolfberry, red dates, poria cocos, konjac powder, and yam is 4-8:0.1-2:0.5-4:0.1-2:0.01-1:7-11:0.1-2:0.5-4:0.5-4; The first mixed bacterial solution consists of: Saccharomyces cerevisiae bacterial solution, Bacillus subtilis bacterial solution, Bacillus licheniformis bacterial solution, Bacillus coagulans bacterial solution, Lactobacillus delbrueckii bacterial solution, Lactobacillus plantarum bacterial solution, Clostridium butyricum bacterial solution and Pediococcus lactis bacterial solution; The second mixed bacterial solution consists of: Saccharomyces cerevisiae bacterial solution, Bacillus coagulans bacterial solution, Clostridium butyricum bacterial solution, and Pediococcus lactis bacterial solution.

2. The method for preparing a feed additive for preventing diarrhea in piglets according to claim 1, characterized in that: In step (1), the amounts of cellulase, hemicellulase, pectinase, and sterilized water used are 5%, 2%, 1%, and 58% of the mass of the mixed Chinese medicine powder, respectively.

3. The method for preparing a feed additive for preventing diarrhea in piglets according to claim 1, characterized in that: In step (1), the amounts of peptone and glucose used are 2% and 4% of the mass of the primary wet powder, respectively.

4. The method for preparing a feed additive for preventing diarrhea in piglets according to claim 1, characterized in that: In step (2), the amounts of Saccharomyces cerevisiae, Bacillus subtilis, Bacillus licheniformis, Bacillus coagulans, Lactobacillus delbrueckii, Lactobacillus plantarum, Clostridium butyricum, and Pediococcus lactis in the first mixed bacterial solution are 1%, 1%, 1%, 2%, 1%, 1%, 1%, and 3% of the fermentation substrate mass, respectively.

5. The method for preparing a feed additive for preventing diarrhea in piglets according to claim 1, characterized in that: In step (3), the amounts of Saccharomyces cerevisiae, Bacillus coagulans, Clostridium butyricum, and Pediococcus lactis in the second mixed bacterial solution are 1%, 2%, 1%, and 3% of the mass of the micronutrient fermentation medium, respectively; the amounts of manganese lactate and copper methionine are 0.2% and 0.15% of the mass of the micronutrient fermentation medium, respectively; the initial culture temperature is 29–31℃, and the culture time is 24–32 h.

6. The method for preparing a feed additive for preventing diarrhea in piglets according to claim 1, characterized in that: In step (3), the amounts of keratinase, glucose, and peptone used are 0.1%, 1%, and 0.2% of the mass of the initial fermentation solution of trace elements, respectively; the re-culturing temperature is 29-31℃, and the culturing time is 24-32h.

7. The method for preparing a feed additive for preventing diarrhea in piglets according to claim 1, characterized in that: In step (4), the amount of micronutrient fermentation liquid used is 10% of the mass of the pre-fermented material; the amounts of *Pediococcus lactis* inoculum and *Saccharomyces cerevisiae* inoculum are 1% and 2% of the mass of the pre-fermented material, respectively; the amounts of cellulase, hemicellulase, pectinase, and galactosidase are 1%, 2%, 0.5%, and 1% of the mass of the pre-fermented material, respectively; and the amount of laccase used is 3% of the mass of the fermented enzymatic hydrolysate.

8. A feed additive prepared by the method of any one of claims 1 to 7.