Composite organic acid for preventing and controlling African swine fever as well as preparation method and application of composite organic acid

Abstract

The invention discloses a composite organic acid for preventing and controlling African swine fever as well as a preparation method and application thereof. The composite organic acid is prepared from the following raw materials: water, carbohydrates, powdery grains, traditional Chinese medicine powder and fermentation mother liquor, the traditional Chinese medicine powder is prepared from astragalus membranaceus, coix seeds, fructus forsythiae, bighead atractylodes rhizome, liquorice and folium isatidis. Compared with the prior art, the traditional Chinese medicine powder components are dialectically selected through traditional Chinese medicines, the amino acid content, the energy content, the crude fat content and the total flavone content of the obtained organic acid are improved through microbial fermentation, the appetite of abnormal pigs or / and diseased pigs is promoted, the physique of the pigs is improved, and infection of the abnormal pigs by the diseased pigs is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine combinations, and in particular to a composite organic acid for preventing and controlling African swine fever, and a preparation method and application thereof. Background Art

[0002] African swine fever (ASF) is an acute, hemorrhagic, and highly contagious disease caused by the African swine fever virus (ASFV). Based on clinical manifestations, it is divided into four main types: the most acute type, the acute type, the subacute type, and the chronic type. The natural incubation period of ASFV is 4-9 days, with the longest incubation period being 21 days. At the onset of the disease, the body temperature rises to 42°C and persists for several days until 48 hours before death, when the body temperature begins to drop. In the early stages of the disease, infected pigs show signs of lethargy, depression, anorexia, and dull eyes. In the middle stages of the disease, external manifestations include redness of the skin, a temperature rise of 40-41°C, and even mucus discharge from the eyes and nose, cyanosis of the ear margins, and rapid breathing. In the late stages of the disease, external manifestations include cyanosis of the skin on the ears, limbs, lower abdomen, and buttocks, as well as ataxia, stiff gait, difficulty breathing, and even death.

[0003] In 2018, the first case of ASF was discovered in Liaoning, China, and African swine fever outbreaks were subsequently found in many places across the country. The aggressive African swine fever outbreak severely impacted China's pig farming industry, and caused drastic fluctuations in China's pig market prices from 2019 to 2022. The prevention and control of ASF has also become a key measure in the production process of the pig farming industry.

[0004] At present, there have been some studies on the use of traditional Chinese medicine to treat African swine fever in China. For example, CN202310121912.3 discloses a traditional Chinese medicine composition for the prevention and control of African swine fever, its preparation method and application. It mainly uses 70-120 parts of bupleurum, 70-120 parts of scutellaria, 170-250 parts of peony, 170-220 parts of artemisia annua, 7-12 parts of mint, 70-120 parts of kudzu root, 50-110 parts of immature bitter orange, 80-110 parts of pinellia, 80-120 parts of isatis root, 8-120 parts of raw licorice, and 8-12 parts of mannan oligosaccharide. It has a good effect on the treatment of ASF by using the dialectical compatibility and mutual assistance of traditional Chinese medicine. However, the invention requires many types of Chinese medicine, and the formulated drugs are ground into powder and mixed with mannan oligosaccharides and then mixed with fertilizers. The mixing results in less release of effective ingredients of the medicinal materials. At the same time, the medicinal materials rely entirely on the pig's stomach to digest and release effective ingredients, resulting in large differences in efficacy and therapeutic effects, thereby causing unstable therapeutic effects. Summary of the Invention

[0005] The purpose of the present invention is to provide a composite organic acid for preventing and controlling African swine fever, which is intended to be used for daily feeding, and has the effect of enhancing the physique and immunity of pigs, playing a role in preventing pigs from contracting African swine fever. When fed during the period of African swine fever, it has a good therapeutic effect on sick pigs.

[0006] To achieve the above object, the present invention provides a composite organic acid for preventing and controlling African swine fever, wherein the raw materials of the composite organic acid include: water, sugar substances, powdered grains, Chinese medicine powder and yeast liquid; The traditional Chinese medicine powder is prepared from astragalus root, coix seed, forsythia suspensa, atractylodes macrocephala, liquorice and isatis indigotica leaf.

[0007] According to the above technical solution, the powdered cereals include one or more of rice flour, corn flour and wheat flour.

[0008] Preferably, in the above technical solution, the raw material ratio of the composite organic acid is 5-7:2-5:10-12:1-4:0.5-2 by weight: water: sugar substance: powdered grain: Chinese medicine powder: yeast solution.

[0009] Preferably, in the above technical solution, the Chinese medicine powder is prepared by mixing the following raw materials in the following weight ratios: The ratio of Astragalus: Coix seed: Forsythia: Atractylodes: Licorice: Isatis indigotica is 6-9:4-6:4-6:3-5:2-5:6-9.

[0010] Preferably, in the above technical solution, the Chinese medicine powder further comprises Salvia miltiorrhiza, Corydalis yanhusuo, Ophiopogon japonicus, and Anemarrhena asphodeloides.

[0011] Preferably, in the above technical solution, the Chinese medicine powder is prepared by mixing the following raw materials in a weight ratio: Astragalus: Coix: Forsythia: Atractylodes: Licorice: Isatis: Salvia: Corydalis: Ophiopogon: Anemarrhena: the ratio is 6-9:4-6:4-6:3-5:2-5:6-9:4-6:6-9:4-6:4-6 Preferably, in the above technical solution, the selection of the fermentation liquid is related to the number of fermentations. When the fermentation number N=1, the yeast solution is made from Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus; When the fermentation number N>1, the fermentation liquid is composed of 10-20% of N-1 complex organic acid.

[0012] Preferably, in the above technical solution, when the fermentation number N>1, the fermentation liquid is composed of 10-20% N-1 complex organic acid, and the content of Lactobacillus plantarum in the N-1 complex organic acid is at least 1×10 7 The content of CFU / g and Rhizopus is at least 1×10 8 CFU / g.

[0013] Preferably, in the above technical solution, the ratio of Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus is 1-3:1-3:3-7 by mass.

[0014] According to the above technical solution, the inventors screened out the fermentation bacteria and their ratio of Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus in a ratio of 1-3:1-3:3-7. The principle is: by studying the characteristics of the fermented material and the requirements of the finished product, Rhizopus was selected to account for the largest proportion. Rhizopus secretes free cellulase during the proliferation and fermentation process, which can catalyze the hydrolysis of cellulose in the raw material, combine with Bacillus amyloliquefaciens to produce amylase, and then decompose the exposed large-molecule starch into small-molecule sugars. Lactobacillus plantarum uses some sugars to ferment to form organic acids and lactic acid, thereby improving the taste and flavor of the complex organic acid.

[0015] Preferably, in the above technical solution, the sugar substance is brown sugar or glucose.

[0016] According to the above technical solution, the principle of using brown sugar or glucose as the sugar substance of the present invention is that brown sugar is a warm food that has the effects of soothing the spleen and liver, strengthening the spleen and warming the stomach. The fermentation and decomposition of edible brown sugar into small molecular components such as amino acids and monosaccharides can strengthen the spleen and soothe the liver. The spleen is the foundation of animal biochemistry, and the liver is an important organ for bile secretion, metabolism, and defense. This can improve the pig's transportation, regulation, and detoxification and defense functions. Glucose provides energy supplements for weaned piglets and promotes the spleen's transportation and transformation functions and the liver's detoxification function.

[0017] A method for preparing the composite organic acid for preventing and controlling African swine fever as described above, the method comprising the following steps: S1, yeast solution preparation: when the fermentation number N=1, Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus are mixed in a preset ratio; When the fermentation number N>1, the fermentation liquid is composed of 10-20% N-1 complex organic acid, and the content of plant lactobacillus in the N-1 complex organic acid is at least 1×10 7 The content of CFU / g and Rhizopus is at least 1×10 8 CFU / g; S2, mixing a sugar substance with water to prepare a sugar solution, wherein the ratio of the sugar substance to water is 5-7:2-5; uniformly mixing the sugar aqueous solution and the yeast solution to obtain a sugar aqueous solution containing yeast solution; S3. Weigh astragalus, coix seed, forsythia, atractylodes, liquorice, isatis indigotica, isatis root, houttuynia cordata, and honeysuckle vine and grind them into traditional Chinese medicine powder. Mix the traditional Chinese medicine powder with powdered grains as dry material. Mix the dry material with the sugar aqueous solution containing the fermentation yeast solution in a ratio of 1:2, seal and ferment to obtain complex organic acid. The fermentation container retains 10-20% of the complex organic acid, and 80-90% of the complex organic acid is used to feed sows or piglets.

[0018] Preferably, in the above technical solution, when the fermentation number N>1, the fermentation liquid is composed of 10-20% N-1 composite organic acid, and when the content of Lactobacillus plantarum in the N-1 composite organic acid is less than 1×10 7 The content of CFU / g or / and Rhizopus is less than 1×10 8 CFU / g, add plant lactobacillus agent or / and Rhizopus agent to the N-1 composite organic acid until the content of plant lactobacillus is at least 1×10 7 CFU / g and the content of Rhizopus was at least 1×10 8 CFU / g.

[0019] An application of the composite organic acid for preventing and controlling African swine fever as described above, wherein abnormal pigs or / and sick pigs are fed the composite organic acid to prevent and control African swine fever in sows or piglets. The principle of the present invention using composite organic acids to prevent and control large-scale infection of African swine fever in sows or piglets is as follows: First, the external characteristics of African swine fever in pigs are mostly: red skin, congestion and cyanosis of the perineum or scrotum, swelling and congestion of the distal limbs or joints, ataxia, stiff gait, poor or no appetite, persistent high fever, diarrhea, bloody stools or vomiting; secondly, pathological autopsy of the sick pigs: enlarged, fragile, dark red or black spleen, kidney bleeding or ecchymosis, enlarged and bleeding lymph nodes in the kidneys and liver, lung bleeding or ecchymosis, and endocardial bleeding. According to the course of African swine fever, the external features and anatomical structure of sick pigs, the symptoms of ASF are analyzed from the perspective of traditional Chinese medicine as follows: (1) The symptoms of sick pigs are red skin, swollen and bleeding spleen, and ecchymosis of kidney. The exogenous epidemic causes water accumulation and heat generation, and dampness and heat coexist in the spleen and kidney, causing spleen and kidney qi stagnation and blood stasis. The spleen is responsible for transportation and transformation and controls the circulation of blood in the veins. Disharmony between the spleen and kidney leads to the body's transportation and transformation disorders, obstructing the skin veins and causing poor blood circulation in the skin, causing congestion of blood vessels on the skin surface; (2) The symptoms of bleeding in the kidneys, liver, spleen, endocardium and other internal organs of sick pigs and high fever and bloody stools are characterized by pathogenic heat entering the blood, the extreme heat in the blood forces the blood to flow abnormally, and the hot blood flows abnormally, which consumes yin blood and vital energy, causing high fever and bleeding.

[0020] Based on the pathology and etiology of African swine fever mentioned above, the inventors proposed a method combining traditional Chinese medicine dialectical diagnosis and treatment methods, including prevention and control. The prevention focuses on strengthening the foundation and nourishing the essence, replenishing qi and raising yang, and nourishing the spleen and kidneys, consolidating and improving the fundamental vitality of the pigs, enhancing the transport function of the spleen and kidneys, promoting the smooth flow of qi and blood in the pigs, eliminating accumulated metabolites in the body, and improving the pigs' resistance to epidemics and external infections; the control focuses on clearing the stranguria to remove dampness and heat, reduce inflammation and reduce fire, cool the blood and eliminate blood stasis, strengthen the spleen and replenish qi, and promote the circulation of qi and blood, so that the qi and blood of the internal organs can be restored and the normal physiological functions can be maintained.

[0021] The principle of the traditional Chinese medicine powder of the present invention for preventing and controlling African swine fever is as follows: coix seed, salvia miltiorrhiza, ophiopogon, rhizoma anemarrhenae and forsythia are used as assistant medicines; rhizoma anemarrhenae clears away heat and detoxifies, forsythia promotes diuresis and relieves stranguria, strengthens the functions of corydalis yanhusuo in promoting blood circulation and qi, and isatis indigotica clears heat and cools blood, removes dampness and heat, and disperses heat evil in the internal organs; forsythia clears the heart and relieves restlessness, and cooperates with astragalus to promote fluid production and nourish blood, soothe the liver and relieve depression, and relieve symptoms of loss of appetite and mental depression; coix seed strengthens the spleen, replenishes qi, and nourishes blood, and ophiopogon nourishes yin and promotes fluid production, strengthens astragalus to strengthen the spleen, tonifies the middle, replenishes qi and raises yang, and regulates the transportation and transformation of the spleen, achieving the effects of strengthening the foundation, strengthening the spleen, replenishing qi, and promoting blood circulation; coix seed clears away heat and relieves stranguria, and is supplemented by isatis indigotica and corydalis to clear heat and reduce fire, fight epidemics and inflammation, strengthen the internal organs to resist the severe qi of the epidemic, and prevent the internal organs from bleeding due to excessive heat and blood. Atractylodes macrocephala is used as an adjuvant and licorice is used as a guiding drug to jointly enhance the functions of the main and auxiliary drugs in nourishing and strengthening the defense, cooling blood and removing blood stasis, resisting epidemics and inflammation, and achieving the effect of eliminating heat evil and resisting epidemic severe qi.

[0022] Compared with the prior art, the present invention has the following beneficial effects: The present invention grinds astragalus, coix seed, forsythia, atractylodes, licorice, salvia miltiorrhiza, corydalis, ophiopogon, anemarrhena and isatis indigotica into powder and ferments them into compound organic acids for daily feeding of pigs based on the pathological characteristics of African swine fever in pigs and the efficacy and mechanism of action of traditional Chinese medicines. The powder is fermented into compound organic acids for daily feeding of pigs, thereby enhancing the immunity of pigs, reducing the risk of pigs contracting the disease and alleviating the severity of the disease after contracting the disease, thereby achieving a preventive effect; the compound organic acids are used to feed confirmed sick pigs. One of the reasons for the aggravation of the symptoms of sick pigs is that the sick pigs eat less or do not eat, resulting in insufficient nutritional supplementation and accelerated physical decline after the disease, and the wet feed with a unique fermented flavor can promote the appetite of sick pigs better than traditional dry feed, thereby alleviating or improving the loss of appetite of sick pigs. The compound organic acids provide sufficient nutritional supply for the sick pigs, and the medicinal materials exert the effects of cooling blood, reducing fire, antibacterial and anti-inflammatory, thereby preventing internal organ bleeding.

[0023] The invention uses a fermentation yeast liquid composed of multiple microorganisms to ferment and prepare composite organic acids. The microorganisms ferment and decompose astragalus, coix seed, forsythia, atractylodes, liquorice, salvia miltiorrhiza, Corydalis yanhusuo, ophiopogon, anemarrhena and isatis indigotica, thereby promoting the release of effective ingredients of traditional Chinese medicines and having better therapeutic effects than traditional powder infiltration feeds. DETAILED DESCRIPTION

[0024] The technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort shall fall within the scope of protection of the present invention. Example 1

[0025] A composite organic acid for preventing and controlling African swine fever, wherein the composite organic acid is prepared from 17.2 L of water, 6.88 kg of brown sugar, 34.4 kg of corn flour, 3.44 kg of traditional Chinese medicine powder, and 1.72 kg of yeast liquid, that is, the composite organic acid raw materials are prepared in a weight ratio of water: sugar substance: powdered grain: traditional Chinese medicine powder: yeast liquid of 5:2:10:1:0.5; The 3.44 kg of traditional Chinese medicine powder is prepared by astragalus: coix seed: forsythia: atractylodes: licorice: isatis indigotica: salvia miltiorrhiza: corydalis: ophiopogon: anemarrhena asphodeloides in a weight ratio of 6:4:4:3:2:6:4:6:4:4, that is, the 3.44 kg of traditional Chinese medicine powder is specifically prepared from 480 g of astragalus, 320 g of coix seed, 320 g of forsythia, 240 g of atractylodes, 160 g of licorice, 480 g of isatis indigotica, 320 g of salvia miltiorrhiza, 480 g of corydalis, 320 g of ophiopogon, and 320 g of anemarrhena; The yeast solution is prepared from Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus in a mass ratio of 1:1:3.

[0026] A method for preparing the composite organic acid for preventing and controlling African swine fever as described above, wherein the steps of the preparation method are as follows: S1, preparation of yeast solution: the fermentation number N is 1, Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus are mixed in a mass ratio of 1:1:3 for the yeast solution, and 20 g of Lactobacillus plantarum, 20 g of Bacillus amyloliquefaciens and 60 g of Rhizopus are weighed and mixed with an aqueous solution to prepare the yeast solution; S2, weigh 6.88 kg of brown sugar and dissolve it in 17.2 L of water to prepare a sugar solution; mixing the fermentation yeast solution with the sugar aqueous solution to obtain a sugar aqueous solution containing fermentation bacteria; S3: Weigh 480 g of astragalus, 320 g of coix seed, 320 g of forsythia, 240 g of atractylodes, 160 g of liquorice, 480 g of isatis indigotica, 320 g of salvia miltiorrhiza, 480 g of corydalis, 320 g of ophiopogon, and 320 g of anemarrhena asphodeloides, respectively, and grind them into 3.44 kg of Chinese medicinal powder. The 3.44 kg of Chinese medicinal powder is mixed with 34.4 kg of corn flour as dry material. The dry material is evenly stirred with a sugar aqueous solution containing fermentation bacteria, and the mixture is sealed and fermented for 3 days to obtain a composite organic acid. 20% of the composite organic acid is retained in the fermentation container, and 80% of the composite organic acid is boiled and sterilized before feeding sows or piglets. Example 2

[0027] A composite organic acid for preventing and controlling African swine fever, wherein the composite organic acid is prepared from 5 L of water, 2 kg of brown sugar, 10 kg of rice flour, 2 kg of traditional Chinese medicine powder, and 1 kg of yeast solution, that is, the composite organic acid raw materials are prepared in a weight ratio of water: sugar substance: powdered grain: traditional Chinese medicine powder: yeast solution of 5:2:10:2:1; The 2 kg of traditional Chinese medicine powder is prepared by mixing astragalus: coix seed: forsythia: atractylodes: licorice: isatis leaf in a weight ratio of 9:6:6:5:5:9, that is, the 2 kg of traditional Chinese medicine powder is prepared by mixing 450 g of astragalus, 300 g of coix seed, 300 g of forsythia, 250 g of atractylodes, 250 g of licorice and 450 g of isatis leaf in a weight ratio of 9:6:6:5:5:9; The content of Lactobacillus plantarum in the 20% complex organic acid retained from the previous fermentation was 3.6×10 7 The content of CFU / g and Rhizopus was 3.5×10 8 CFU / g, so 1 kg of complex organic acid retained from the previous fermentation was used as the yeast liquid.

[0028] A method for preparing the composite organic acid for preventing and controlling African swine fever as described above, wherein the steps of the preparation method are as follows: S1, yeast liquid preparation: This fermentation is the Nth fermentation, and N>1, take part of the N-1th composite organic acid to test the content of Lactobacillus plantarum and Rhizopus in the composite organic acid, and the test result is that the content of Lactobacillus plantarum is 3.6×10 7 The content of CFU / g and Rhizopus was 3.5×10 8 CFU / g, 1 kg of N-1 compound organic acid was used as the yeast liquid; S2, weigh 2 kg of brown sugar and dissolve it in 10 L of water to prepare a sugar solution; Take the above sugar aqueous solution and mix it evenly with 1 kg of N-1 composite organic acid to obtain a sugar aqueous solution containing fermentation bacteria; S3: Weigh 450 g of astragalus, 300 g of coix seed, 300 g of forsythia, 250 g of atractylodes, 250 g of liquorice, and 450 g of isatis indigotica respectively, mix and grind into Chinese medicinal powder, mix the Chinese medicinal powder with 10 kg of rice flour as dry material, stir the dry material evenly with a sugar aqueous solution containing fermentation bacteria, and ferment in a sealed container for 3 days to obtain composite organic acid. 20% of the composite organic acid is retained in the fermentation container, and 80% of the composite organic acid is taken for boiling and sterilization and then used to feed sows or piglets. Example 3

[0029] A composite organic acid for preventing and controlling African swine fever, wherein the composite organic acid is prepared from 9.45 L of water, 6.75 kg of brown sugar, 16.2 kg of wheat flour, 5.4 kg of traditional Chinese medicine powder, and 2.7 kg of yeast liquid, that is, the composite organic acid raw materials are prepared in a weight ratio of 7:5:12:4:2 of water: sugar: powdered grain: traditional Chinese medicine powder: yeast liquid; Wherein, the 5.4 kg of traditional Chinese medicine powder is prepared by astragalus: coix seed: forsythia: atractylodes: licorice: isatis indigotica: salvia miltiorrhiza: corydalis: ophiopogon: anemarrhena in a weight ratio of 7:5:5:4:4:7:5:7:5:5, that is, the 5.4 kg of traditional Chinese medicine powder is specifically prepared by 700 g of astragalus, 500 g of coix seed, 500 g of forsythia, 400 g of atractylodes, 400 g of licorice, 700 g of isatis indigotica, 500 g of salvia miltiorrhiza, 700 g of corydalis, 500 g of ophiopogon and 500 g of anemarrhena; The content of Lactobacillus plantarum and Rhizopus in 10% composite organic acid was detected, and the content of Lactobacillus plantarum was 3.5×10 6 The content of CFU / g and Rhizopus was 4.1×10 8 CFU / g, 2.7 kg of complex organic acids retained from the previous fermentation were used as the yeast liquid.

[0030] A method for preparing the composite organic acid for preventing and controlling African swine fever as described above, wherein the steps of the preparation method are as follows: S1, yeast liquid preparation: This fermentation is the Nth fermentation, and N>1, take part of the N-1th composite organic acid to test the content of Lactobacillus plantarum and Rhizopus in the composite organic acid, and the test result is that the content of Lactobacillus plantarum is 3.5×10 6 CFU / g and the content of Rhizopus was 4.1×10 8 CFU / g, so 2.7 kgN-1 compound organic acid was used as the yeast liquid; S2, weigh 6.75 kg of brown sugar and dissolve it in 9.45 L of water to prepare a sugar solution; Taking the above sugar solution and mixing it with the fermentation yeast solution to obtain a sugar solution containing fermentation bacteria; S3, respectively weigh 700 g of astragalus, 500 g of coix seed, 500 g of forsythia, 400 g of atractylodes, 400 g of liquorice, 700 g of isatis indigotica, 500 g of salvia miltiorrhiza, 700 g of corydalis, 500 g of ophiopogon and 500 g of anemarrhena, mix and grind into Chinese medicinal powder, mix the Chinese medicinal powder with 16.2 kg of wheat flour as dry material, stir the dry material with the sugar-containing aqueous solution, seal and ferment for 4 days to obtain composite organic acid, obtain composite organic acid, retain 20% of the composite organic acid in the fermentation container, and take 80% of the composite organic acid, boil and sterilize it, and then feed it to sows or piglets.

[0031] Comparative Example 1 A traditional Chinese medicine composition for preventing and controlling African swine fever, the traditional Chinese medicine composition is prepared from 480 g of Radix Astragali, 320 g of Coix Seed, 320 g of Forsythia, 240 g of Atractylodes Macrocephala Koidz, 160 g of Glycyrrhiza, 480 g of Dandelion, 320 g of Salvia Miltiorrhiza, 480 g of Corydalis, 320 g of Ophiopogon and 320 g of Anemarrhena Asphodeloides Bunge; A preparation method of a traditional Chinese medicine composition for preventing and controlling African swine fever, the preparation method is as follows: S1, 480 g of Radix Astragali, 320 g of Coix Seed, 320 g of Forsythia, 240 g of Atractylodes Macrocephala Koidz, 160 g of Glycyrrhiza, 480 g of Dandelion, 320 g of Salvia Miltiorrhiza, 480 g of Corydalis, 320 g of Ophiopogon and 320 g of Anemarrhena Asphodeloides Bunge are weighed and ground into powder respectively; S2, the traditional Chinese medicine powder is mixed with dry feed (adult pig feed) and used for feeding sows or piglets.

[0032] Comparative Example 2 The same as Example 1, the difference is that the fermentation broth is prepared from Lactobacillus plantarum and Clostridium butyricum at a mass ratio of 1:1.

[0033] A preparation method of a complex organic acid for preventing and controlling African swine fever as described above, the steps of the preparation method are as follows: S1, fermentation broth preparation: fermentation frequency N=1, the fermentation broth is prepared from Lactobacillus plantarum and Bacillus amyloliquefaciens at a mass ratio of 1:1, 20 g of Lactobacillus plantarum and 20 g of Bacillus amyloliquefaciens are weighed and mixed to prepare a fermentation inoculum.

[0034] Comparative Example 3 The same as Example 1, the difference is that the fermentation broth is prepared from Bacillus subtilis, yeast and Rhizopus at a mass ratio of 1:1:3.

[0035] A preparation method of a complex organic acid for preventing and controlling African swine fever as described above, the steps of the preparation method are as follows: S1, fermentation broth preparation: this time the fermentation frequency N is 1, the fermentation broth is prepared from lactic acid bacteria, yeast and Aspergillus at a mass ratio of 1:1:3, 20 g of lactic acid bacteria, 20 g of yeast and 60 g of Aspergillus are weighed and mixed to prepare a fermentation inoculum.

[0036] Comparative Example 4 The same as Example 1, the difference is that the fermentation broth is prepared from 100 g of Rhizopus.

[0037] A preparation method of a complex organic acid for preventing and controlling African swine fever as described above, the steps of the preparation method are as follows: S1, yeast liquid preparation: fermentation number N = 1, weigh 100 g of Rhizopus and mix well to make yeast liquid.

[0038] Comparative Example 5 The same as Example 2, except that the yeast solution is N-1 times compound organic acid, and the content of Lactobacillus plantarum in the N-1 times compound organic acid solution is 2.6×10 6 The content of CFU / g and Rhizopus was 4.7×10 7 CFU / g.

[0039] Comparative Example 6 It is basically the same as Example 2, except that the components of the Chinese medicine powder are different.

[0040] The content of Lactobacillus plantarum in the N-1 composite organic acid solution of this embodiment is 3.8×10 7 CFU / g and the content of Rhizopus was 4.0×10 8 CFU / g, meeting the requirement of N-1 times composite organic acid solution as yeast solution, the medicinal powder in this embodiment is obtained by preparing 480 g of astragalus, 320 g of coix seed, 320 g of isatis root, 240 g of white atractylodes, 160 g of liquorice and 480 g of scutellaria.

[0041] Comparative Example 7 It is basically the same as Example 2, except that the components of the Chinese medicine powder are different.

[0042] The content of Lactobacillus plantarum in the N-1 composite organic acid solution of this embodiment is 3.8×10 7 CFU / g and the content of Rhizopus was 4.0×10 8 CFU / g, meeting the requirement of N-1 times composite organic acid solution as yeast solution, the medicinal powder in this embodiment is obtained by preparing 450 g of astragalus, 300 g of coix seed, 250 g of licorice, 450 g of isatis indigotica, 320 g of ophiopogon and 320 g of anemarrhena.

[0043] The composite organic acids or substances prepared by the methods of Examples 1-3 and Comparative Examples 1-7 and then sterilized by boiling for use in feeding pigs were tested for microbial contaminant hygiene according to GB 13078-2017. The results are shown in Table 1 below.

[0044] Table 1 Microbial content of each example name Total mold count (CFU / g) Total bacterial count (CFU / g) Salmonella (CFU / g) Example 1 <![CDATA[5.2×10 2 ]]> <![CDATA[1.8×10 3 ]]> Not detected Example 2 <![CDATA[4.4×10 2 ]]> <![CDATA[2.1×10 3 ]]> Not detected Example 3 <![CDATA[7.1×10 2 ]]> 3.2 x 10 3 ]]> Not detected Comparative Example 1 1.2×10 <![CDATA[2.8×10 2 ]]> Not detected Comparative Example 2 4.6 x 10 2 ]]> <![CDATA[2.3×10 4 ]]> Not detected Comparative Example 3 <![CDATA[4.2×10 2 ]]> <![CDATA[3.5×10 3 ]]> Not detected Comparative Example 4 <![CDATA[2.3×10 2 ]]> <![CDATA[2.4×10 2 ]]> Not detected Comparative Example 5 6.2 x 10 2 ]] 4.1 x 10 3 ]]> Not detected Comparative Example 6 <![CDATA[3.7×10 2 ]]> <![CDATA[3.9×10 3 ]]> Not detected Comparative Example 7 <![CDATA[4.4×10 2 ]]> 2.6 x 10 3 ]]> Not detected It can be concluded from Table 1 that the total number of molds in the composite organic acids or substances used to feed pigs after boiling sterilization in Examples 1-3 and Comparative Examples 1-7 is less than 4×10 4 CFU / g, total bacterial count less than 2×10 6CFU / g, no Salmonella was detected, it meets the GB 13078-2017 hygiene standards and can be used for pig feeding.

[0045] The nutritional components of the composite organic acids or substances prepared by the methods of Examples 1-3 and Comparative Examples 1-7 and then sterilized by boiling for feeding pigs are shown in Table 2. The total flavonoid content is determined by the following method: (1) Preparation of standard curve: Accurately weigh 10.0 mg of rutin reference substance and dilute to 10 mL with 30% ethanol to obtain 1.0 mg / mL standard solution. Add 0, 0.5, 1, 1.5, 2 and 2.5 mL of standard solution to a 10 mL volumetric flask, respectively, to prepare 2.5, 2.0, 1.5, 1.0, 0.5 and 0 mL of 30% ethanol solution, respectively. Then add 0.5 mL of 5% sodium nitrite, shake well, and let it stand for 6 minutes. Add 0.5 mL of 10% aluminum nitrate solution, let it stand for 6 minutes, add 4.0 mL of 4% sodium hydroxide solution, and dilute to the scale with distilled water. After shaking well and letting it stand for 15 minutes, measure the absorbance at 510 nm. Prepare a standard curve with absorbance as the vertical axis and total flavonoids concentration as the vertical axis; (2) Extraction and determination of total flavonoids in samples: Accurately weigh 3.0 g of the sample to be tested and place it in a 200 mL Soxhlet extraction flask. Add 90 mL of 70% ethanol and extract at 80°C for 3 h. The volume is then fixed in a 100 mL volumetric flask. Accurately pipette 2 mL of the sample extract into a 10 mL volumetric flask, add 0.5 mL of 5% sodium nitrite solution, shake well, and let it stand for 6 min. Add 0.5 mL of 10% aluminum nitrate solution and let it stand for 6 min. Add 4.0 mL of 4% sodium hydroxide solution and shake well. Add 30% ethanol to the mark, let it stand for 15 min, and measure the absorbance at 510 nm. Calculate the total flavonoid content in the sample based on the fitted standard curve.

[0046] Table 2 Nutritional composition of the composite organic acids prepared in various examples name Gross energy (MJ / kg) Crude fat (%) calcium(%) Total phosphorus (%) Total amino acids (%) Total flavonoids content (mg / g) Example 1 46.36 34.47 0.33 0.57 21.26 20.14 Example 2 45.22 33.61 0.30 0.49 20.23 17.77 Example 3 47.14 36.28 0.36 0.53 20.12 19.56 Comparative Example 1 35.62 20.71 0.26 0.43 12.74 —— Comparative Example 2 43.64 27.31 0.35 0.50 15.36 15.21 Comparative Example 3 43.16 26.82 0.34 0.56 16.23 17.94 Comparative Example 4 40.38 23.59 0.33 0.47 13.23 14.38 Comparative Example 5 43.36 27.57 0.36 0.54 15.26 14.87 Comparative Example 6 41.24 34.38 0.35 0.52 20.17 18.89 Comparative Example 7 40.91 33.29 0.33 0.53 20.34 16.75 Abnormal pigs were separated into pens, and throat swabs were collected from the abnormal pigs. The abnormal adult pigs were characterized by: decreased feed intake, loss of appetite, significantly decreased feed intake, lying down more and moving less, a body temperature of 40-41°C, cyanosis of the ear edges, or redness of the skin. The throat swabs were sent for testing, and sick pigs that tested positive for HZ antigen were separated into pens again, and the original pens were disinfected. A total of 128 abnormal pigs and 42 sick pigs were fed with the composite organic acids of Example 1 and Comparative Examples 1-7, respectively. The abnormal pigs (86 pigs) and sick pigs (42 pigs) were fed for 30 days. Their feed intake, body temperature, and disease changes were observed. Throat swabs were collected on the 10th and 30th days for HZ antigen testing. The results are shown in Tables 3 and 4.

[0047] Table 3 Conditions of sick pigs after administration of compound organic acids name Number Body temperature Feeding and activity HZ antigen positive on day 10 (only) HZ antigen positive on day 30 (only) Example 1 7 Body temperature returns to normal on the 4th to 6th day On days 8-9, food intake and activity levels increase 2 0 Comparative Example 1 5 Body temperature returns to normal on the 10th to 15th day Days 17-18: Increased food intake and activity 5 1 Comparative Example 2 5 Body temperature returns to normal on the 9th to 12th day Days 12-15: Increased food intake and activity 3 0 Comparative Example 3 5 Body temperature returns to normal on the 8th to 11th day Days 8-13: Increased food intake and activity 2 0 Comparative Example 4 5 Body temperature returns to normal on the 12th to 14th day Days 14-16: Increased food intake and activity 3 0 Comparative Example 5 5 Body temperature returns to normal on the 8th to 10th day Days 15-18: Increased food intake and activity 2 0 Comparative Example 6 5 Body temperature returns to normal on the 10th to 13th day Days 12-16: Increased food intake and activity 4 2 Comparative Example 7 5 Body temperature returns to normal on the 14th to 16th day Days 18-19: Increased food intake and activity 3 1 The symptoms of sick pig No. 5 in Example 1 group before and after treatment are as follows: Symptoms before treatment: preferred to lie down, would not be driven away, refused to eat dry feed, had red ear margins, body temperature of 42°C, and a throat swab test was positive for HZ antigen; The compound organic acid fermented in Example 1 was fed daily, and the following results were observed: 1-3 days: No dry feed, but low intake of compound organic acids, prefers to lie down, does not move even when driven away, ears are red and cyanotic, body temperature is 42°C, and throat swab HZ antigen test is positive; Day 4-6: The baby's complex organic acids increase gradually, he likes to lie down, drives away activity, his ears turn red and cyanotic, his temperature is 40°C, and a throat swab HZ antigen test is positive. Days 7-9: There is a slight excess of complex organic acids, appetite increases, lying down decreases, activity increases, the area of ​​cyanosis around the ear margin decreases, body temperature is 38°C, and a throat swab HZ antigen test is negative on day 9; Days 10-15: 200 g of compound organic acid was added to each meal without any excess, activity increased, cyanosis of the ear margins gradually resolved, body temperature was 38.5°C, and a throat swab HZ antigen test was negative; 15-30 days: 300 g of complex organic acid was added to each meal without any residue, activity level returned to normal, ear margins returned to normal, body temperature was 38.5℃, and the throat swab HZ antigen test was negative.

[0048] During the 30-day feeding process, the abnormal pigs (86 pigs) were observed for symptom changes, and throat swabs were collected every 3-5 days to detect HZ antigen.

[0049] Table 4 Pigs fed for 2 weeks name Number Feeding and activity New HZ positive test within 10 days (individuals) New HZ positive test within 20 days (individuals) New HZ positive test within 30 days (individuals) Example 1 11 Increased food intake and decreased lying down on days 3-7 0 0 0 Comparative Example 1 11 On days 11-13, food intake increased and lying down decreased 2 1 1 Comparative Example 2 11 Days 9-12: Increased food intake, unchanged activity 1 1 0 Comparative Example 3 11 Days 8-12: Increased food intake and activity 1 1 0 Comparative Example 4 11 Days 12-16: Increased food intake and activity 1 1 0 Comparative Example 5 11 Days 10-13: Increased food intake and activity 1 0 0 Comparative Example 6 10 Days 10-13: Increased food intake and activity 2 2 1 Comparative Example 7 10 Days 12-16: Increased food intake and activity 2 1 1 The symptoms of abnormal pig No. 10 in Example 1 group before and after treatment are as follows: Symptoms before treatment: preferred lying down, moved after being driven, ate little dry feed, vomited, had diarrhea, body temperature was 40°C, and the throat swab HZ antigen test was negative; The compound organic acid fermented in Example 1 was fed daily, and the following results were observed: Days 1-3: The patient consumes complex organic acids, prefers to lie down, is driven to move, does not vomit, has diarrhea, has a body temperature of 40.5°C, and a throat swab HZ antigen test is negative; Day 4-6: Consume compound organic acids, move around more and lie down less, do not vomit or have diarrhea, temperature is 38.5°C, and throat swab HZ antigen test is negative; 8-9 days: each meal added 100 g of complex organic acid without remainder, activity increased significantly, body temperature detection was 38℃, throat swab HZ antigen detection was negative; 10-15 days: each meal added 200 g of complex organic acid without remainder, activity increased significantly, body temperature detection was 38.5℃, throat swab HZ antigen detection was negative; 15-30 days: each meal added 500 g of complex organic acid without remainder, activity returned to normal, body temperature detection was 38℃, throat swab HZ antigen detection was negative.

[0050] The symptoms of No. 3 sick pig in the comparative example 7 group before and after treatment are as follows: Symptoms before treatment: not eating, lying down, stiff moving front limbs, red ear margin, body temperature detection was 41℃, throat swab HZ antigen detection was negative; Each day, the pig was fed with the complex organic acid fermented according to the ratio of comparative example 7, and the observation results were as follows: 1-3 days: little complex organic acid was eaten, lying down, stiff moving front limbs, red ear margin, body temperature detection was 41.5℃, throat swab HZ antigen detection was negative; 4-6 days: there was much complex organic acid left, lying down, stiff moving front limbs, red ear margin, body temperature detection was 40℃, throat swab HZ antigen detection was negative; 7-9 days: there was much complex organic acid left, lying down decreased, stiff moving front limbs recovered, red ear margin faded, body temperature detection was 39℃, throat swab HZ antigen detection was negative; 10-12 days: there was little complex organic acid left, activity increased, body temperature recovered, ear margin returned to normal color, body temperature detection was 38.5℃, throat swab HZ antigen detection was negative; 12-17 days: each meal added 100 g of complex organic acid without remainder, activity increased significantly, body temperature detection was 38.5℃, throat swab HZ antigen detection was negative; 18-30 days: each meal added 300 g of complex organic acid without remainder, activity returned to normal, body temperature detection was 38℃, throat swab HZ antigen detection was negative.

[0051] The symptoms of No. 8 abnormal pig in the comparative example 7 group before and after treatment are as follows: Symptoms before treatment: not eating, lying down, moving, red ear margin, vomiting, body temperature detection was 40℃, throat swab HZ antigen detection was negative; Each day, the pig was fed with the complex organic acid fermented according to the ratio of comparative example 7, and the observation results were as follows: 1-3 days: little complex organic acid was eaten, lying down, not moving, red ear margin, not vomiting, diarrhea, body temperature detection was about 40℃, throat swab HZ antigen detection was negative; Days 4-6: The child eats less complex organic acids, prefers to lie down, and will not move unless driven away. The ear margins are red and cyanotic, and the child has diarrhea. On the fourth day, the temperature is 42°C, and a throat swab test for HZ antigen is positive. 7-9 days: Eat less complex organic acids, like to lie down, and have poor forelimb movement. The cyanosis of the ear edges subsides, and the body temperature drops to around 40°C. The throat swab HZ antigen test is positive. 10-14 days: There is a surplus of complex organic acids, but the baby's food intake increases with each meal. The baby likes to lie down and moves after a long time. The ear edges turn red and the body temperature drops to around 39°C. On the 14th day, the HZ antigen test of the throat swab is negative. Days 15-17: No residual complex organic acids, increased activity, body temperature of 38.0°C, and a throat swab HZ antigen test negative; 18-30 days: 100 g of complex organic acid was added to each meal without any residue, activity level increased, body temperature was 38.5℃, and throat swab HZ antigen test was negative.

[0052] The invention can be implemented in various ways and is not limited to the embodiments described above. A person skilled in the art will appreciate that the invention can be implemented in other specific ways without changing the technical concept or essential features of the invention. Therefore, it should be understood that the embodiments described above are illustrative and not intended to limit the invention.

Claims

1. A composite organic acid for preventing and controlling African swine fever, characterized in that: The raw materials of the composite organic acid include: water, sugar, powdered grains, Chinese medicinal powder and yeast liquid; The traditional Chinese medicine powder is prepared from astragalus root, coix seed, forsythia suspensa, atractylodes macrocephala, liquorice and isatis indigotica leaf.

2. The composite organic acid for preventing and controlling African swine fever according to claim 1, characterized in that: Calculated by weight, the raw material ratio of the composite organic acid is 5-7:2-5:10-12:1-4:0.5-2: water: sugar: powdered grains: Chinese medicinal powder: yeast liquid.

3. The composite organic acid for preventing and controlling African swine fever according to claim 1, characterized in that: The Chinese medicinal powder further comprises salvia miltiorrhiza, corydalis yanhusuo, ophiopogon japonicus and anemarrhena asphodeloides.

4. The composite organic acid for preventing and controlling African swine fever according to claim 3, characterized in that: It is prepared by mixing the following raw materials in the following weight ratios: The ratio of Astragalus: Coix seed: Forsythia: Atractylodes: Licorice: Isatis indigotica: Salvia miltiorrhiza: Corydalis: Ophiopogon japonicus: Anemarrhena asphodeloides is 6-9:4-6:4-6:3-5:2-5:6-9:4-6:6-9:4-6:4-6.

5. The composite organic acid for preventing and controlling African swine fever according to claim 1, characterized in that: The selection of the fermentation liquid is related to the number of fermentations N. When the fermentation number N=1, the yeast solution is made from Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus; When the fermentation number N>1, the fermentation liquid is composed of 10-20% of N-1 complex organic acid.

6. The composite organic acid for preventing and controlling African swine fever according to claim 5, characterized in that: When the fermentation number N>1, the fermentation liquid is composed of 10-20% N-1 complex organic acid, and the content of plant lactobacillus in the N-1 complex organic acid is at least 1×10 7 The CFU / g content of Rhizopus was at least 1×10 8 CFU / g.

7. The composite organic acid for preventing and controlling African swine fever according to claim 5, characterized in that: Calculated by mass, the ratio of Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus is 1-3:1-4:3-8.

8. A method for preparing a composite organic acid for preventing and controlling African swine fever according to any one of claims 1 to 7, characterized in that: The method steps include: S1, yeast solution preparation: when the fermentation number N=1, Lactobacillus plantarum, Bacillus amyloliquefaciens and Rhizopus are mixed in a preset ratio; When the fermentation number N>1, the fermentation liquid is composed of 10-20% N-1 complex organic acid, and the content of plant lactobacillus in the N-1 complex organic acid is at least 1×10 7 The CFU / g content of Rhizopus was at least 1×10 8 CFU / g; S2, mixing a sugar substance with water to prepare a sugar solution, wherein the ratio of the sugar substance to water is 5-7:2-5; uniformly mixing the sugar aqueous solution and the fermentation yeast solution to obtain a sugar aqueous solution containing fermentation bacteria; S3, weighing astragalus, coix seed, forsythia, atractylodes, liquorice, isatis indigotica, isatis root, houttuynia cordata, and honeysuckle vine and grinding them into traditional Chinese medicine powder, mixing the traditional Chinese medicine powder with powdered grains as dry material, mixing the dry material with the sugar aqueous solution containing fermentation bacteria in a ratio of 1:2, and sealing and fermenting to obtain complex organic acid, retaining 10-20% of the complex organic acid in the fermentation container, and using 80-90% of the complex organic acid for feeding sows or piglets.

9. The method for preparing a composite organic acid for preventing and controlling African swine fever according to claim 8, characterized in that: When the fermentation number N>1, the fermentation liquid is composed of 10-20% N-1 composite organic acid, and when the content of plant lactobacillus in the N-1 composite organic acid is less than 1×10 7 The content of CFU / g or / and Rhizopus is less than 1×10 8 CFU / g, add plant lactobacillus agent or / and Rhizopus agent to the N-1 composite organic acid until the content of plant lactobacillus is at least 1×10 7 CFU / g and the content of Rhizopus was at least 1×10 8 CFU / g.

10. A use of the composite organic acid for preventing and controlling African swine fever according to any one of claims 1 to 7, characterized in that: Feed abnormal pigs and / or sick pigs with compound organic acids to prevent and control African swine fever in sows or piglets.