Bacillus velezensis SXDYYB2 and application thereof in prevention and treatment of white scour of piglets

Abstract

The invention belongs to the technical field of prevention and treatment of white scour of piglets, relates to bacillus velezensis SXDYYB2 and application of the bacillus velezensis SXDYYB2 in prevention and treatment of white scour of piglets, provides bacillus velezensis SXDYYB2 with the preservation number of CCTCC NO: M2025257, and further provides a composition for prevention and treatment of white scour of piglets, and the composition is prepared from the following dry medicinal materials in parts by weight: 20-30 parts of fructus mume, 2-3 parts of fructus forsythiae, 2-3 parts of fructus forsythiae, 2-3 parts of fructus forsythiae, 2-3 parts of fructus forsythiae, 2-3 parts of fructus forsythiae, 2-3 parts of fructus forsythiae and 2-3 parts of fructus forsythiae. The feed additive is prepared from the following raw materials in parts by weight: 10-20 parts of radix glycyrrhizae, 20-30 parts of herba taraxaci, 10-30 parts of rhizoma coptidis and 1-3 parts of an SXDYYB2 microbial preparation. The traditional Chinese medicine composition is prepared by boiling smoked plum, fructus forsythiae, liquorice, dandelion and coptis chinensis in water at normal temperature to obtain liquid medicine and then adding an SXDYYB2 microbial preparation. The composition for preventing and treating the white scour of the piglets, disclosed by the invention, can play a relatively good role in preventing and treating the white scour of the piglets.

Description

Technical Field

[0001] The invention belongs to the technical field of piglet pullorum prevention and treatment, and relates to Bacillus velezensis SXDYYB2 and application thereof in preventing and treating piglet pullorum. Background Art

[0002] Pork is a key indicator of improving people's living standards and a significant factor influencing price fluctuations. The key to the development of the pig farming industry lies in piglets. There was once a saying, "Raising good piglets is half the battle," and strong piglets contribute to a high meat production rate. Piglets are most susceptible to pullorrhoea (a disease caused by Escherichia coli) after birth. Affected pigs are mostly under five days old. Initially, one or two pigs die suddenly, and then the entire litter is affected. Affected pigs often become listless, huddle together, and pass yellow, pulpy feces. They quickly become emaciated, eventually falling into a coma and dying. Piglets with pullorrhoea are emaciated and unwilling to eat. Initial diarrhea progresses to pullorrhoea, which is contagious and can lead to death if not treated promptly. Puerorrhoea in piglets requires prevention, but currently, there are no effective preventive or therapeutic drugs. Chemical antibiotics have both preventative and therapeutic effects on pullorrhoea in piglets and are generally the first choice for treatment. However, Escherichia coli, the pathogen of piglet pullorum, is very likely to develop resistance to chemical antibiotics such as gentamicin, enrofloxacin, and fluphenazine. Even if multiple antimicrobial drugs are used alternately or in combination for treatment or prevention in a planned manner, the development of resistance cannot be completely avoided.

[0003] In the treasure house of traditional Chinese medicine, there are many Chinese herbal medicines with good disease resistance. The main mechanisms of these Chinese herbal medicines against piglet white diarrhea are as follows: (1) Inhibition of pathogens: Some Chinese herbal medicines and their components can directly inhibit the pathogens that cause piglet white diarrhea. For example, studies have shown that Chinese herbal medicines such as Sophora flavescens have no or only weak inhibitory effects on toxigenic Escherichia coli, which means that they may work through other mechanisms. (2) Antagonism of enterotoxins: Extracts of Chinese herbal medicines such as honeysuckle and forsythia have antagonistic effects on Escherichia coli heat-sensitive enterotoxin (LT), which is an important mechanism for their treatment of piglet white diarrhea. This antagonistic effect can help alleviate the diarrhea symptoms caused by enterotoxins. (3) Inhibition of enterotoxin production: Some Chinese herbal medicines such as honeysuckle, forsythia and water pepper can inhibit the production of LT by Escherichia coli, which is also one of their mechanisms for treating piglet white diarrhea. By reducing the production of enterotoxins, the inflammatory response and diarrhea degree of the piglet intestine can be reduced. (4) Nonspecific anti-diarrhea effect: Chinese herbs such as Sophora flavescens, Cyperus multiflorus, and Polygonum hydropiper also have nonspecific anti-diarrhea effects, which may be an important part of their medicinal mechanism for treating piglet scours. This effect may be related to regulating intestinal motility, protecting the intestinal mucosa, or improving the balance of intestinal microorganisms.

[0004] The mechanisms of action of Chinese herbal medicine in treating pullorum in piglets are multifaceted, including direct inhibitory effects on pathogens and enterotoxins, as well as nonspecific pathways to improve intestinal function and reduce diarrheal symptoms. These mechanisms work together to achieve a therapeutic effect.

[0005] Probiotics are living microorganisms that have a beneficial effect on the human body by improving the balance of intestinal flora. They are also known as microecological preparations. They have developed rapidly in the past 10 years and are a research hotspot in nutrition. Their main function is to help establish a gastrointestinal microbial flora that is beneficial to the host through the competitive exclusion of digestive tract organisms, prevent bacterial and viral diarrhea, and promote growth. Their main mechanism of action is: (1) Competitive inhibition: Probiotics can prevent the colonization and reproduction of pathogens (such as Escherichia coli) by occupying ecological sites on the surface of the intestinal mucosa. This competitive inhibition helps to reduce the number of pathogens, thereby reducing the incidence of piglet white diarrhea. (2) Enhance immune function: Probiotics can stimulate the immune system of piglets and enhance their resistance to pathogens. They can promote the activity of immune cells and increase the production of antibodies, thereby improving the overall immunity of piglets. (3) Regulate the balance of intestinal microecology: Probiotics can help maintain the balance of normal intestinal flora, inhibit the growth of harmful bacteria, and promote the reproduction of beneficial bacteria. This balance helps maintain intestinal health and prevent the occurrence of diseases such as diarrhea. (4) Improve the intestinal environment: Probiotics produce organic acids (such as lactic acid and acetic acid) through metabolism. These organic acids can lower the intestinal pH value, creating an environment that is not conducive to the growth of pathogens. In addition, probiotics can also produce some antibacterial substances that directly inhibit the growth of pathogens. Promote nutrient absorption: Probiotics can help piglets better digest and absorb nutrients, strengthen their physical fitness, and thus improve their resistance to disease.

[0006] Although Chinese herbal medicines and probiotics can improve the intestinal microecological environment of piglets and enhance their immunity, for serious infections that have already occurred, especially piglet white diarrhea caused by Escherichia coli, the use of Chinese herbal medicines or probiotics alone may not achieve the ideal prevention and control effect. Sometimes the piglets die from severe diarrhea before the medicine takes effect, thus losing the medicine's due effect. Summary of the Invention

[0007] In order to solve the above technical problems, the present invention provides a Bacillus Velez subtilis SXDYYB2 and its application in preventing and treating piglet pullorum. The composition of the microbial preparation including Bacillus Velez subtilis SXDYYB2 can play a good preventive and therapeutic role on piglet pullorum.

[0008] The technical solution of the present invention to solve the above technical problems is as follows: The present invention provides a Bacillus velezinis ( Bacillus velezensis)SXDYYB2, the deposit number is CCTCCNO:M2025257.

[0009] The present invention also provides the above-mentioned Bacillus Velezii ( Bacillus velezensis ) Application of SXDYYB2 in preventing and treating white diarrhea in piglets.

[0010] Furthermore, the application includes using the Bacillus Velezii ( Bacillus velezensis )SXDYYB2 and / or its metabolites.

[0011] The present invention also provides a composition for preventing and treating piglet pullorum, the composition comprising the above-mentioned Bacillus Velezii ( Bacillus velezensis )SXDYYB2 microbial preparation made from SXDYYB2.

[0012] Furthermore, the composition is composed of the following dry medicinal raw materials: black plum, forsythia, liquorice, dandelion, coptis, and SXDYYB2 microbial preparation.

[0013] Ume: Latin name Mume Fruit ; It tastes sour and astringent, is neutral in nature, and enters the liver, spleen, lung, and large intestine meridians; it is bitter and cold, and has a diuretic and stranguria-relieving effect, and can break up blood and dredge the menstruation. Its effects: astringes the lungs, astringes the intestines, promotes the production of body fluids, and calms ascaris.

[0014] Forsythia suspensa: Latin name Forsythia hanging ; It is a heat-clearing and detoxifying medicine with bitter and slightly cold nature, and belongs to the lung, heart, and small intestine meridians; when used, it can be used together with honeysuckle, dandelion, etc. Modern pharmacological research shows that this medicine has antibacterial, antipyretic, antiemetic, anti-inflammatory, diuretic, anti-liver damage, cardiotonic, diuretic, and antihypertensive effects. It has a certain inhibitory effect on Salmonella typhi, Salmonella paratyphi, Escherichia coli, Shigella dysenteriae, diphtheria, cholera, Staphylococcus aureus, Streptococcus, etc.

[0015] Licorice: Latin name Licorice Root ; It is flat in nature, sweet in taste, with a slight smell and a sweet and special taste; it is mainly used to clear away heat and detoxify, eliminate phlegm and relieve cough, and relieve abdominal distension, etc.

[0016] Dandelion: Latin name Dandelion Herb ; sweet, slightly bitter, cold; clears heat and detoxifies, reduces swelling and resolves stagnation.

[0017] Coptis chinensis: Latin name Coptis Rhizome ; It tastes extremely bitter, clears away heat and dampness, and relieves fire and detoxifies.

[0018] SXDYYB2 has strong protease, lipase, amylase and other activities, can produce antibiotics, has strong biological oxygen-snatching ability in the animal intestine, and can produce L-lactic acid under anaerobic conditions. These characteristics play an important role in promoting the digestion and absorption of animal nutrients, improving the feed conversion rate of animals, preventing diseases and promoting growth.

[0019] Furthermore, the composition is composed of the following dry medicinal materials in parts by weight: 20-30 parts of black plum, 2-3 parts of forsythia, 10-20 parts of licorice, 20-30 parts of dandelion, 10-30 parts of coptis, and 1-3 parts of SXDYYB2 microbial preparation.

[0020] Furthermore, the preparation method of the SXDYYB2 microbial preparation is as follows: the Bacillus velezensis ( Bacillus velezensis ) SXDYYB2 seed liquid is inoculated into a fermentation medium, and after continuous culture at 30-37°C for 24-72 hours, it is statically cultured for 96-120 hours to obtain a fermentation liquid; the fermentation liquid is centrifuged to collect the bacteria, and after freeze-drying, the SXDYYB2 microbial preparation is obtained.

[0021] Furthermore, the fermentation medium is composed of the following components in parts by weight: 1-5 parts of peptone, 1-3 parts of beef extract, 1-10 parts of yeast extract, 5-15 parts of sodium chloride, 10-100 parts of fructooligosaccharides, and 1000 parts of water.

[0022] Further preferably, the fermentation medium consists of the following components in parts by weight: 3 parts of peptone, 2 parts of beef extract, 5 parts of yeast extract, 10 parts of sodium chloride, 50 parts of fructooligosaccharides, and 1000 parts of water.

[0023] The present invention also provides a method for preparing the above-mentioned composition for preventing and treating pullorum in piglets, comprising the following preparation steps: 1) Weigh the dried medicinal materials (Prunus mume, Forsythia suspensa, Licorice root, Taraxacum officinale, and Coptis chinensis) by weight, grind them, and pass them through an 80-mesh sieve. Add 10-30 times the total weight of the medicinal materials in room temperature water and boil for 30 minutes. Filter to obtain a medicinal solution. 2) After cooling the medicinal solution obtained in step 1), the SXDYYB2 microbial preparation is added to obtain the traditional Chinese medicine composition.

[0024] Furthermore, the bacterial count of the SXDYYB2 microbial preparation is greater than or equal to 1.80×10 9 CFU / mL.

[0025] More preferably, the bacterial count of the SXDYYB2 microbial preparation is 1.80×10 9-10 CFU / mL.

[0026] The beneficial effects of the present invention are as follows: the composition for treating piglet pullorum is prepared by boiling black plum, forsythia, licorice, dandelion, and coptis root at room temperature to obtain a medicinal liquid, and then adding the SXDYYB2 microbial preparation; the black plum (containing organic acids and flavonoids) regulates intestinal pH and inhibits the growth of pathogenic Escherichia coli (ETEC), while also providing a suitable microenvironment for probiotics (SXDYYB2) and enhancing their colonization ability; the forsythia (containing forsythiaside and forsythiaside) has a broad-spectrum antibacterial effect, reduces harmful intestinal bacteria, and synergistically reduces the diarrhea rate of piglets with SXDYYB2; Licorice (glycyrrhizic acid and glycyrrhetinic acid) fights inflammation, repairs intestinal mucosal damage, and alleviates intestinal edema caused by diarrhea in piglets, while also enhancing the immunomodulatory function of SXDYYB2. Dandelion and Coptis chinensis (chlorogenic acid and berberine) synergistically inhibit ETEC toxin secretion and reduce excessive intestinal water secretion (coptis berberine blocks bacterial adhesion, while dandelion polysaccharides promote the proliferation of probiotics). SXDYYB2 microbial preparations directly and competitively inhibit pathogenic bacterial colonization, secrete antimicrobial peptides (such as surfactin), and, together with traditional Chinese medicine ingredients, regulate intestinal microecological balance. Feeding sows with this composition mixed with their feed before farrowing can prevent the disease. After birth, feeding piglets with this composition mixed with their feed can prevent the disease. After the onset of the disease, feeding them with the composition can cure the disease completely, with both prevention and cure rates exceeding 70%.

[0027] Biological Deposit Description Biological material: SXDYYB2, taxonomic name: Bacillus velezinis ( Bacillus velezensis ), deposited in the China Center for Type Culture Collection on February 19, 2025. The address of the collection is: Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province; the collection number is CCTCC No: M 2025257. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a microscopic photograph of the Bacillus velezensis SXDYYB2 of the present invention. DETAILED DESCRIPTION

[0029] The principles and features of the present invention are described below (in conjunction with the accompanying drawings). The examples given are only used to explain the present invention and are not used to limit the scope of the present invention.

[0030] The present invention discloses a Bacillus velezensis SXDYYB2 and its application in preventing and treating piglet pullorum. Those skilled in the art can refer to the contents of this article and appropriately improve the process parameters to achieve the desired effect. It should be noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0031] It should be understood that the expression "one or more of" includes individually each of the items recited after the expression and various combinations of two or more of the recited items, unless otherwise apparent from the context and usage. The expression "and / or" in conjunction with three or more recited items should be understood to have the same meaning, unless otherwise apparent from the context.

[0032] The terms "comprising", "having" or "containing", including their grammatical synonyms, should generally be understood as open and non-restrictive, e.g., not excluding other unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0033] It should be understood that the order of steps or the order in which certain actions are performed is not important as long as the application remains operable. Additionally, two or more steps or actions may be performed simultaneously.

[0034] The use of any and all examples or exemplary language such as "for example" or "including" herein is intended only to better illustrate the present application and does not limit the scope of the present application. No language in this specification should be construed as indicating any non-claimed element is essential to the practice of the present application.

[0035] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0036] The sources of some raw materials of the present invention are as follows: Black plum: purchased from Anhui Bozhou Traditional Chinese Medicine Market, according to the 2020 edition of the "Chinese Pharmacopoeia", the citric acid content is 18.7%; Forsythia: purchased from Sichuan Xinhehua Traditional Chinese Medicine Pieces Co., Ltd. (batch number: SC2023-05), the forsythin content is ≥0.25%.

[0037] The present invention provides a Bacillus velezinis ( Bacillus velezensis )SXDYYB2, the deposit number is CCTCCNO:M2025257.

[0038] The present invention also provides the above-mentioned Bacillus Velezii ( Bacillus velezensis ) Application of SXDYYB2 in preventing and treating white diarrhea in piglets.

[0039] As an optional embodiment, the application includes using the Bacillus Velezii ( Bacillus from Velez )SXDYYB2 and / or its metabolites.

[0040] The present invention also provides a composition for preventing and treating piglet pullorum, the composition comprising the above-mentioned Bacillus Velezii ( Bacillus velezensis )SXDYYB2 microbial preparation made from SXDYYB2.

[0041] As an optional embodiment, the composition is composed of the following dry medicinal raw materials or dried medicinal raw materials: black plum, forsythia, liquorice, dandelion, coptis and SXDYYB2 microbial preparation.

[0042] The black plum (containing organic acids and flavonoids) of the present invention regulates intestinal pH, inhibits the growth of pathogenic Escherichia coli (ETEC), and at the same time provides a suitable microenvironment for probiotics (SXDYYB2), thereby enhancing their colonization ability; Forsythia (including forsythiaside and forsythiaside) has a broad-spectrum antibacterial effect, reduces harmful intestinal flora, and synergizes with SXDYYB2 to reduce the diarrhea rate in piglets; Licorice (glycyrrhizic acid, glycyrrhetinic acid) is anti-inflammatory, repairs intestinal mucosal damage, relieves intestinal wall edema caused by diarrhea in piglets, and enhances the immune regulation function of SXDYYB2; Dandelion and Coptis chinensis (chlorogenic acid, berberine) synergistically inhibit ETEC toxin secretion and reduce excessive intestinal water secretion (e.g., Coptis chinensis berberine can block bacterial adhesion, and dandelion polysaccharides promote the proliferation of probiotics); The SXDYYB2 microbial preparation directly and competitively inhibits the colonization of pathogenic bacteria, secretes antimicrobial peptides (such as surfactin), and simultaneously regulates the balance of intestinal microecology together with traditional Chinese medicine ingredients.

[0043] As an optional embodiment, the composition is composed of the following dry medicinal materials in parts by weight: 20-30 parts of black plum, 2-3 parts of forsythia, 10-20 parts of licorice, 20-30 parts of dandelion, 10-30 parts of coptis, and 1-3 parts of SXDYYB2 microbial preparation.

[0044] As an optional embodiment, the preparation method of the SXDYYB2 microbial preparation is: the Bacillus Velezii ( Bacillus velezensis) SXDYYB2 seed liquid is inoculated into a fermentation medium, and after continuous culture at 30-37°C for 24-72 hours, it is statically cultured for 96-120 hours to obtain a fermentation liquid; the fermentation liquid is centrifuged to collect the bacteria, and after freeze-drying, the SXDYYB2 microbial preparation is obtained.

[0045] Specifically, the fermentation broth was centrifuged at 4°C and 8000 rpm (r / min) for 15 minutes, the bacterial solution was collected, sterilized skimmed milk powder was used as a freeze-drying protective agent, the bacterial solution was pre-cooled at -20°C for 24 hours, and then placed in a vacuum freeze dryer at -50°C to a moisture content of 3-5%, thereby obtaining the SXDYYB2 microbial preparation.

[0046] As an optional embodiment, the fermentation medium is composed of the following components in parts by weight: 1-5 parts of peptone, 1-3 parts of beef extract, 1-10 parts of yeast extract, 5-15 parts of sodium chloride, 10-100 parts of fructooligosaccharides, and 1000 parts of water.

[0047] Further preferably, the fermentation medium consists of the following components in parts by weight: 3 parts of peptone, 2 parts of beef extract, 5 parts of yeast extract, 10 parts of sodium chloride, 50 parts of fructooligosaccharides, and 1000 parts of water.

[0048] The present invention also provides a method for preparing the above-mentioned composition for preventing and treating pullorum in piglets, comprising the following preparation steps: 1) Weigh the dried medicinal materials (Prunus mume, Forsythia suspensa, Licorice root, Taraxacum officinale, and Coptis chinensis) by weight, grind them, and pass them through an 80-mesh sieve. Add 10-30 times the total weight of the medicinal materials in room temperature water and boil for 30 minutes. Filter to obtain a medicinal solution. 2) After cooling the medicinal solution obtained in step 1), the SXDYYB2 microbial preparation is added to obtain the traditional Chinese medicine composition.

[0049] As an optional embodiment, the bacterial count of the SXDYYB2 microbial preparation is greater than or equal to 1.80×10 9 CFU / mL.

[0050] Further preferably, the bacterial count of the SXDYYB2 microbial preparation is 1.80×109-10CFU / mL.

[0051] The present invention is described in detail below in conjunction with embodiments and experimental data.

[0052] Culture medium formula: Luria-Bertani agar (LB) nutrient medium: peptone 10.0 g / L, sodium chloride 5.0 g / L, yeast extract 5.0 g / L.

[0053] Example 1 (1) Isolation and screening of strains: The SXDYYB2 strain of the present invention was isolated from the intestine of a healthy piglet and identified as Bacillus velezinis by 16S rRNA. Specifically, the SXDYYB2 strain was isolated from the intestinal contents of a healthy piglet aged 7-14 days and obtained by primary screening with LB medium (containing 0.1% bile salts). The 16S rRNA and gyrB genes were identified as Bacillus velezinis ( Bacillus velezensis ).

[0054] (2) Strain preservation: The SXDYYB2 strain was deposited in the China Center for Type Culture Collection (CCTCC, address: Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province, Postal Code: 430072) on February 19, 2025, and was classified and named Bacillus velezensis ( Bacillus velezensis ), the deposit number is CCTCC No: M 2025257.

[0055] (3) Evaluation of strain characteristics: The diameter of the inhibition zone of this strain against ETEC reached 18.5±1.2 mm (Oxford cup method), and the number of viable ETEC cells could be reduced by 3.2 log after 6 h of co-culture. 10 CFU / mL, with typical Bacillus morphological characteristics, Gram staining positive, catalase and VP test positive, and no hemolysis, no virulence and drug resistance genes, fully meeting the standards for feed probiotics.

[0056] Example 2 The composition for preventing and treating piglet pullorum in this embodiment is prepared from the following dry medicinal materials in parts by weight: 20 parts of black plums, 2 parts of forsythia suspensa, 10 parts of liquorice, 20 parts of dandelions, 10 parts of coptis chinensis, and 1 part of SXDYYB2 microbial preparation.

[0057] The preparation method of the probiotic fermentation Chinese medicine composition for preventing and treating piglet pullorum of this embodiment comprises the following steps: 1) Weigh the dried herbs (Prunus mume, Forsythia suspensa, Licorice root, Taraxacum officinale, and Coptis chinensis) by weight, crush (pass through an 80-mesh sieve), add 30 times the total weight of the herbs in room temperature water, and boil for 30 minutes; 2) After cooling the medicinal solution obtained in step 1), the SXDYYB2 microbial preparation is added to obtain the traditional Chinese medicine composition.

[0058] The SXDYYB2 microbial preparation is prepared by inoculating SXDYYB2 seed liquid into a culture medium for fermentation; the fermentation conditions are: continuous culture at 35°C for 48 hours and then static culture for 96 hours to obtain a fermentation liquid; the fermentation liquid is centrifuged at 4°C and 8000 rpm (r / min) for 15 minutes to collect the bacterial liquid, sterilized skim milk powder is used as a freeze-drying protective agent, the bacterial liquid is pre-cooled at -20°C for 24 hours, and then placed in a vacuum freeze dryer at -50°C to a moisture content of 3-5%, thereby obtaining a SXDYYB2 microbial preparation containing more than 2×10 9 CFU / mL; the culture medium consists of the following components by weight: 3 parts of peptone, 2 parts of beef extract, 7 parts of yeast extract, 11 parts of sodium chloride, 75 parts of fructooligosaccharide, and 1000 parts of water.

[0059] Example 3 The composition for preventing and treating piglet pullorum in this embodiment is prepared from the following dry medicinal materials in parts by weight: 25 parts of black plums, 3 parts of forsythia suspensa, 15 parts of liquorice, 26 parts of dandelions, 18 parts of coptis chinensis, and 2 parts of SXDYYB2 microbial preparation.

[0060] The preparation method of the composition for preventing and treating piglet pullorum of this embodiment comprises the following steps: 1) Weigh the dried herbs (Prunus mume, Forsythia suspensa, Licorice root, Taraxacum officinale, and Coptis chinensis) by weight, crush (pass through an 80-mesh sieve), add 30 times the total weight of the herbs in room temperature water, and boil for 30 minutes; 2) After cooling the medicinal solution obtained in step 1), the SXDYYB2 microbial preparation is added to obtain the traditional Chinese medicine composition.

[0061] The SXDYYB2 microbial preparation is prepared by inoculating SXDYYB2 seed liquid into a culture medium for fermentation; the fermentation conditions are: continuous culture at 35°C for 48 hours and then static culture for 96 hours to obtain a fermentation liquid; the fermentation liquid is centrifuged at 4°C and 8000 rpm (r / min) for 15 minutes to collect the bacterial liquid, sterilized skim milk powder is used as a freeze-drying protective agent, the bacterial liquid is pre-cooled at -20°C for 24 hours, and then placed in a vacuum freeze dryer at -50°C to a moisture content of 3-5%, thereby obtaining a SXDYYB2 microbial preparation containing more than 2×10 9 CFU / mL; the culture medium consists of the following components by weight: 3 parts of peptone, 2 parts of beef extract, 7 parts of yeast extract, 11 parts of sodium chloride, 75 parts of fructooligosaccharide, and 1000 parts of water.

[0062] Example 4 The composition for preventing and treating piglet pullorum in this embodiment is prepared from the following dry medicinal materials in parts by weight: 30 parts of black plums, 3 parts of forsythia suspensa, 20 parts of liquorice, 30 parts of dandelions, 30 parts of coptis chinensis, and 3 parts of SXDYYB2 microbial preparation.

[0063] The preparation method of the composition for preventing and treating piglet pullorum of this embodiment comprises the following steps: 1) Weigh the dried herbs (Prunus mume, Forsythia suspensa, Licorice root, Taraxacum officinale, and Coptis chinensis) by weight, crush (pass through an 80-mesh sieve), add 30 times the total weight of the herbs in room temperature water, and boil for 30 minutes; 2) After cooling the medicinal solution obtained in step 1), the SXDYYB2 microbial preparation is added to obtain the traditional Chinese medicine composition.

[0064] The SXDYYB2 microbial preparation is prepared by inoculating SXDYYB2 seed liquid into a culture medium for fermentation; the fermentation conditions are: continuous culture at 35°C for 48 hours and then static culture for 96 hours to obtain a fermentation liquid; the fermentation liquid is centrifuged at 4°C and 8000 rpm (r / min) for 15 minutes to collect the bacterial liquid, sterilized skim milk powder is used as a freeze-drying protective agent, the bacterial liquid is pre-cooled at -20°C for 24 hours, and then placed in a vacuum freeze dryer at -50°C to a moisture content of 3-5%, thereby obtaining a SXDYYB2 microbial preparation containing more than 2×10 9 CFU / mL; the culture medium consists of the following components by weight: 3 parts of peptone, 2 parts of beef extract, 7 parts of yeast extract, 11 parts of sodium chloride, 75 parts of fructooligosaccharide, and 1000 parts of water.

[0065] Comparative Example 1 The composition for preventing and treating piglet pullorum in this comparative example is prepared from the following dry medicinal materials in parts by weight: 3 parts of Forsythia suspensa, 15 parts of Licorice, 26 parts of Taraxacum mongolicum, 18 parts of Coptis chinensis, and 2 parts of SXDYYB2 microbial preparation.

[0066] The preparation method of the composition for preventing and treating piglet pullorum in this comparative example comprises the following steps: 1) Weigh the dried herbs (Fructus Forsythiae, Licorice, Taraxacum Herba, and Coptis Chinensis) by weight, crush (pass through an 80-mesh sieve), add 30 times the total weight of the herbs in room temperature water, and boil for 30 minutes; 2) After cooling the medicinal solution obtained in step 1), the SXDYYB2 microbial preparation is added to obtain the traditional Chinese medicine composition.

[0067] The SXDYYB2 microbial preparation is prepared by inoculating SXDYYB2 seed liquid into a culture medium for fermentation; the fermentation conditions are: continuous culture at 35°C for 48 hours and then static culture for 96 hours to obtain a fermentation liquid; the fermentation liquid is centrifuged at 4°C and 8000 rpm (r / min) for 15 minutes to collect the bacterial liquid, sterilized skim milk powder is used as a freeze-drying protective agent, the bacterial liquid is pre-cooled at -20°C for 24 hours, and then placed in a vacuum freeze dryer at -50°C to a moisture content of 3-5%, thereby obtaining a SXDYYB2 microbial preparation containing more than 2×10 9 CFU / mL; the culture medium consists of the following components by weight: 3 parts of peptone, 2 parts of beef extract, 7 parts of yeast extract, 11 parts of sodium chloride, 75 parts of fructooligosaccharide, and 1000 parts of water.

[0068] Comparative Example 2 Compared with Example 3, the only difference is that Forsythia suspensa was not added.

[0069] Comparative Example 3 Compared with Example 3, the only difference is that no dandelion is added.

[0070] Comparative Example 4 Compared with Example 3, the only difference is that no SXDYYB2 microbial preparation is added.

[0071] Comparative Example 5 Compared with Example 3, the difference is that only the SXDYYB2 microbial preparation is contained.

[0072] Comparative Example 6 Compared with Example 3, the only difference is that commercially available probiotics (commercially available lactobacillus preparation) are used instead of the SXDYYB2 microbial preparation.

[0073] The following experimental examples illustrate the beneficial effects of the present invention. Experimental methods in the following experimental examples where specific conditions are not specified are generally measured in accordance with national standards. If no corresponding national standards exist, the methods are conducted in accordance with commonly used international standards, conventional conditions, or conditions recommended by the manufacturer.

[0074] Experimental Example 1 Antibacterial Experiment (1) Prepare the culture medium plate (MH agar plate): Prepare the hydrolyzed casein agar medium according to the product instructions, sterilize it at 121 °C for 15 min until the temperature drops to about 55 °C, and pour it into a sterile plate to maintain the agar thickness of 4 mm; (2) Escherichia coli (O_(128) serotype Escherichia coli) was diluted with physiological saline. The dilution standard was 1.5×10 5 CFU / mL concentration, and then MH agar plate smearing and inoculation operations were performed. The plate was covered and placed in a clean bench and dried for more than 3 minutes, with a maximum time of 15 minutes. The alcohol lamp was then used to ignite the plate, and the agar and the composition were mixed. During this period, two sterilized filter papers were covered on the Oxford cup, the cup lid was closed, and the plate was placed horizontally. The plate was incubated in a 37°C constant temperature incubator for about 22 hours. The test results were observed and the diameter of the inhibition zone was measured. The measurement results are shown in Table 1 below: Table 1 Antibacterial test results

[0075] It can be seen from Table 1 that the composition of the present invention (SXDYYB2 microbial preparation, black plum, forsythia, licorice, dandelion, and coptis chinensis) has the best antibacterial effect. The antibacterial effect of the composition of Comparative Example 4 lacking the SXDYYB2 microbial preparation of the present invention is poor. The antibacterial effect of the commercially available lactobacillus preparation in Comparative Example 6 after combination with traditional Chinese medicine is not as good as that of the composition of the present invention, which proves that the SXDYYB2 microbial preparation of the present invention cooperates with black plum, forsythia, licorice, dandelion, and coptis chinensis to achieve a better antibacterial effect.

[0076] Experimental Example 2: Treatment Effect Experiment Experimental animals: Piglets diagnosed with pullorum disease.

[0077] Confirmed piglets were randomly divided into an infection group (piglets with pullorum disease, untreated) and an experimental group (piglets with pullorum disease, treated). The experimental groups were treated with the compositions of Examples 2-4 and Comparative Examples 1-6, respectively. Each piglet was fed 100 grams of the composition mixed with 1000 grams of feed per meal for three consecutive days, followed by a seven-day interval before starting another course of treatment, for a total of three consecutive courses. The infected piglets ate and drank normally, and their clinical changes were observed daily. Confirmed piglets were considered cured when their symptoms disappeared and they returned to normal. The cure rate was calculated, and the experimental results are shown in Table 2 below: Table 2 Treatment effect results

[0078] As can be seen from Table 2, the compositions of Examples 2-4 have a high cure rate for sick piglets, reaching a cure rate of 90%. Compared with Comparative Example 4 (without SXDYYB2 microbial preparation), Comparative Example 5 (only SXDYYB2 microbial preparation), and Comparative Example 6 (commercially available lactobacillus preparation and traditional Chinese medicine), the cure rate is higher, proving that the SXDYYB2 microbial preparation of the present invention cooperates with black plum, forsythia, licorice, dandelion, and coptis chinensis to achieve a better therapeutic effect.

[0079] Experimental Example 3: Preventive Effect Experiment The composition of the present invention was used to conduct a preventive experiment on farrowing sows and unmarketed piglets: Prevention of sows: The sows were divided into a blank group (no prevention) and a prevention group (prevention). The prevention groups were respectively treated with the compositions of Examples 2-4 and Comparative Examples 1-6. 100 g of the composition was mixed with 3000 g of feed per meal and fed to the sows for 3 days. The disease condition of the piglets after delivery was observed, and the prevention rate was calculated.

[0080] Prevention of unsold piglets; the unsold piglets were divided into a blank group (no prevention) and a prevention group (prevention). The prevention groups were respectively treated with the compositions of Examples 2-4 and Comparative Examples 1-6, and 20 grams per piglet was mixed into the feed and fed for 3 consecutive days. The disease condition of the slaughtered piglets was observed, and the prevention rate was calculated. Prevention rate / %=number of uninfected pigs / number of normal pigs in each group*100%.

[0081] The results of the preventive effect are shown in Table 3 below: Table 3 Prevention effect results

[0082] As can be seen from the above table, the compositions of Examples 2-4 of the present invention have a higher preventive effect, with the highest prevention rate reaching 86.6%. Compared with the blank group and the prevention group fed with the compositions of Comparative Examples 1-6, the prevention rate is significantly higher, indicating that the compositions of the present invention have a better preventive effect.

[0083] In summary, the composition of the present invention can play a good preventive and therapeutic effect on piglet pullorum. Before the sow gives birth, the composition of the present invention is mixed with feed and fed to the sow, which can prevent piglet pullorum; after the piglet is born, before pullorum occurs, the composition of the present invention is mixed with feed and fed to the piglet, which can prevent piglet pullorum; after the piglet becomes ill, the composition of the present invention is fed to the piglet, which can cure the disease, and the prevention and cure rates can reach more than 70%.

[0084] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A Bacillus velezinoffii ( Bacillus velezensis )SXDYYB2, characterized in that, The deposit number is CCTCCNO:M2025257.

2. A Bacillus Velezii as claimed in claim 1 ( Bacillus velezensis ) Application of SXDYYB2 in preventing and treating white diarrhea in piglets.

3. The use according to claim 2, characterized in that The application includes using the Bacillus Velezii ( Bacillus velezensis )SXDYYB2 and / or its metabolites.

4. A composition for preventing and treating piglet pullorum, characterized in that: The composition comprises the Bacillus Velezii ( Bacillus velezensis )SXDYYB2 microbial preparation made from SXDYYB2.

5. A composition for preventing and treating piglet pullorum according to claim 4, characterized in that: The composition is composed of the following dry medicinal raw materials: black plum, forsythia, liquorice, dandelion, coptis root, and SXDYYB2 microbial preparation.

6. A composition for preventing and treating piglet pullorum according to claim 4, characterized in that: The composition is composed of the following dry medicinal materials in parts by weight: 20-30 parts of black plums, 2-3 parts of forsythia suspensa, 10-20 parts of liquorice, 20-30 parts of dandelions, 10-30 parts of coptis chinensis, and 1-3 parts of SXDYYB2 microbial preparation.

7. A composition for preventing and treating piglet pullorum according to claim 5, characterized in that: The preparation method of the SXDYYB2 microbial preparation is as follows: Bacillus velezensis ) SXDYYB2 seed liquid is inoculated into a fermentation medium, and after continuous culture at 30-37°C for 24-72 hours, it is statically cultured for 96-120 hours to obtain a fermentation liquid; the fermentation liquid is centrifuged to collect the bacteria, and after freeze-drying, the SXDYYB2 microbial preparation is obtained.

8. A composition for preventing and treating piglet pullorum according to claim 7, characterized in that: The fermentation medium consists of the following components in parts by weight: 1-5 parts of peptone, 1-3 parts of beef extract, 1-10 parts of yeast extract, 5-15 parts of sodium chloride, 10-100 parts of fructooligosaccharide, and 1000 parts of water.

9. The method for preparing the composition for preventing and treating pullorum in piglets according to any one of claims 5 to 8, wherein: The method comprises the following preparation steps: 1) Weigh the dried medicinal materials (Prunus mume, Forsythia suspensa, Licorice root, Taraxacum officinale, and Coptis chinensis) by weight, grind them, and pass them through an 80-mesh sieve. Add 10-30 times the total weight of the medicinal materials in room temperature water, boil for 30 minutes, and filter to obtain a medicinal solution. 2) After cooling the medicinal solution obtained in step 1), the SXDYYB2 microbial preparation is added to obtain the traditional Chinese medicine composition.

10. The method for preparing the composition for preventing and treating pullorum in piglets according to claim 9, wherein: The bacterial count of the SXDYYB2 microbial preparation is greater than or equal to 1.80×10 9 CFU / mL.

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