WO2016127956A1 - 丁酸梭菌ucn-12菌株及其组合物和应用 - Google Patents

丁酸梭菌ucn-12菌株及其组合物和应用 Download PDF

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WO2016127956A1
WO2016127956A1 PCT/CN2016/078912 CN2016078912W WO2016127956A1 WO 2016127956 A1 WO2016127956 A1 WO 2016127956A1 CN 2016078912 W CN2016078912 W CN 2016078912W WO 2016127956 A1 WO2016127956 A1 WO 2016127956A1
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clostridium butyricum
ucn
group
strain
fermentation culture
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French (fr)
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郑一敏
曾雁鸣
周永喜
胥秀英
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成都与康科技有限公司
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/145—Clostridium
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor

Definitions

  • the invention belongs to the field of microbiology and relates to a strain of Clostridium butyricum UCN-12 and compositions and uses thereof.
  • Clostridium perfringens also known as gastrobacteria, is one of the most common diseases of food-induced gastroenteritis.
  • Foreign statistics caused by food poisoning caused by Clostridium perfringens accounted for about 30% of bacterial food poisoning. The latest statistics show that the number of food poisoning caused by Clostridium perfringens in the United States is about 250,000 per year, resulting in economic losses of 120 million. Dollar.
  • Epidemiological investigations have found that an average of every outbreak of food poisoning caused by Clostridium perfringens usually involves 50-100 people. Some serious food poisoning outbreaks involve even thousands of people.
  • Clostridium welchii is generally divided into six types A, B, C, D, E and F, and type A and type E bacteria and ⁇ produced by them are the main pathogenic factors.
  • the pathogen invades the body from the digestive tract or wound, and multiplies in the small intestine and cecal chorion to produce a strong alpha toxin, which changes the permeability of the capillaries, and causes the toxin to enter the blood of the livestock and a large amount, causing systemic toxemia.
  • Clostridium is a common disease in livestock and poultry. Every year, a large number of animals die from different types of Clostridium perfringens infection. The disease has an acute onset, short course of disease, sudden death without any pre-symptoms, and is endemic or sporadic. It can occur all year round, especially in winter and spring, and the incidence rate is 100% in severe cases. The mortality rate is extremely high, reaching 95%, which is an acute infectious disease that seriously harms the production of rabbits.
  • Clostridium perfringens mainly uses cephalosporin, ampicillin, penicillin, chloramphenicol, gentamicin and tetracycline. Due to the drug resistance caused by long-term use of antibiotics, the use of antibiotics is getting higher and higher, the therapeutic effect is getting worse and worse, and the environmental pollution is getting bigger and bigger. The safety of special drug residues is becoming more and more a concern of the whole society. The problem. Vaccine is a good method to prevent Clostridium perfringens, but the protection period is short, and the cost is high, which is difficult to accept in production.
  • Chinese Patent Application No. 201210390182.9 discloses a veterinary medicine for preventing and treating Wilson's disease of piglets and a preparation method thereof, the formula comprising 10-20 parts of berberine, 10-20 parts of astragalus, 5-15 parts of forsythia, honeysuckle 5-15 parts, 10-20 parts of angelica, 10-20 parts of white peony, 1-5 parts of notoginseng powder, 5-15 parts of ginseng stems and leaves and 5-10 parts of licorice, this method is difficult to eliminate people's concerns about drug residues And the cost is high.
  • Patent application 201210552595.2 discloses a special feed containing Bacillus licheniformis, Bacillus subtilis, Enterococcus faecium, Streptococcus faecalis, xylanase, due to the interaction between Bacillus licheniformis and Bacillus subtilis and Enterococcus faecium and Streptococcus faecalis, The compatibility of the composition is not reasonable, and the method is not effective.
  • Patent application 201310577204.7 and 201210261894.0 respectively disclose a strain of Bacillus subtilis and an acid-resistant, heat-resistant and bile-tolerant Bacillus licheniformis to control Clostridium perfringens, the strain used is completely different from the present invention, and at the same time, Bacillus subtilis And Bacillus licheniformis is an aerobic bacterium, requires aeration culture, the process is complicated, and the production cost is also higher than the present invention.
  • the first technical problem to be solved by the present invention is to provide a new strain of Clostridium butyricum, the fermentation culture of the strain of the present invention, the fermentation liquid or the bacterial body obtained by centrifugation of the fermentation culture, and the fermentation liquid or the bacterial body.
  • the dried product is used as an active ingredient to feed the animal. It has the treatment and/or prevention of Wilson's disease in livestock and poultry. No antibiotics are added in the whole process, and the antibiotic residue is avoided to affect human health. The environmental pollution caused by the extensive use of antibiotics in animal breeding is completely eliminated. .
  • the new strain of the present invention is Clostridium butyricum, numbered UCN-12, deposited on September 15, 2014 at the General Microbiology Center (CGMCC) of the China Microbial Culture Collection Management Committee, and the deposit number is CGMCC No.9652. . Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Beichen West Road, Chaoyang District, Beijing 100101, China. It was converted from domestic deposit to international deposit on March 10, 2016.
  • CGMCC General Microbiology Center
  • the 16S rDNA sequence of the Clostridium butyricum UCN-12 strain is shown in SEQ ID No. 1.
  • Clostridium butyricum UCN-12 strain of the present invention is different from the previously reported strains of Clostridium butyricum.
  • the 16s rDNA sequence there are also differences in biochemical properties, as shown in Table 1. The most obvious difference is that the amount of hexanoic acid produced by Clostridium butyricum UCN-12 is more than 10 times that of butyric acid, sometimes 20 times, showing a characteristic flavor of caproic acid, while the existing Clostridium butyricum does not produce caproic acid.
  • a second technical problem to be solved by the present invention is to provide a composition comprising the Clostridium butyricum UCN-12 strain of the present invention as a main active ingredient, which can be used as a feed additive.
  • the composition comprises a prophylactically and/or therapeutically effective amount of a Clostridium butyricum UCN-12 strain, a fermentation culture of a Clostridium butyricum UCN-12 strain, and the culture solution obtained by centrifuging the fermentation culture or a dried product thereof, The bacterial body obtained by centrifugation of the fermentation culture or a dried product thereof is used as an active ingredient, and an acceptable auxiliary material is added.
  • Acceptable excipients are commercially acceptable carriers or forms for animal feed Agent.
  • the composition is in unit dosage form, preferably wherein the unit dosage form is selected from the group consisting of: tablets, granules, powders, oral solutions, oral suspensions, oral emulsions and premixes, and the like.
  • a third technical problem to be solved by the present invention is to provide a fermentation culture of Clostridium butyricum UCN-12 strain, Clostridium butyricum UCN-12 strain, a culture solution obtained by centrifuging the fermentation culture or a dried product thereof.
  • the cells obtained by centrifugation of the fermentation culture or the dried product thereof are used for the preparation of a veterinary drug, a feed additive and/or a feed for treating or preventing Wilson's disease in livestock and poultry.
  • the fourth technical problem to be solved by the present invention is a fermentation culture of Clostridium butyricum UCN-12 strain, Clostridium butyricum UCN-12 strain, a culture solution obtained by centrifugation of the fermentation culture or a dried product thereof, and the fermentation culture
  • the cells obtained by centrifugation or the dried product thereof are used for the preparation of veterinary drugs, feed additives and/or feeds for reducing cholesterol and fat in eggs, removing the taste of eggs and improving the flavor of eggs.
  • the inventors have found that not only can the cholesterol and fat in the eggs be significantly reduced, but it is also found that after feeding the strain of Clostridium butyricum UCN-12 of the present invention and its derivatives, the taste of the eggs can be completely removed, and the egg flavor exceeds that of the wild-raised eggs. And the taste is better.
  • the above-mentioned effects of the novel strain of the invention not only improve the health quality of the cultured animal products, but also increase the added value and production efficiency of the product market.
  • the fifth technical problem solved by the present invention is a fermentation culture of Clostridium butyricum UCN-12 strain, Clostridium butyricum UCN-12 strain, a culture solution obtained by centrifugation of the fermentation culture or a dried product thereof, and the fermentation culture
  • the bacteria obtained by centrifugation or the dried product thereof are used in veterinary drugs, feed additives and/or feeds for improving the production performance of livestock and poultry, and high-yield animal breeding is realized.
  • the sixth technical problem solved by the present invention is a fermentation culture of Clostridium butyricum UCN-12 strain, Clostridium butyricum UCN-12 strain, a culture solution obtained by centrifugation of the fermentation culture or The dried product, the cells obtained by centrifugation of the fermentation culture or the dried product thereof are used for the preparation of a veterinary drug, a feed additive and/or a feed for treating or preventing prenatal and postpartum constipation in a sow.
  • the seventh technical problem solved by the present invention is a fermentation culture of Clostridium butyricum UCN-12 strain, Clostridium butyricum UCN-12 strain, a culture solution obtained by centrifugation of the fermentation culture or a dried product thereof, and the fermentation culture
  • the bacteria obtained by centrifugation or the dried product thereof are used for preparing veterinary drugs, feed additives and/or feeds for improving the smell of livestock and poultry; the odor of the feces of the cultured animals is significantly reduced, and the adverse effects of animal feces on the environment are reduced.
  • Environmentally friendly is a fermentation culture of Clostridium butyricum UCN-12 strain, Clostridium butyricum UCN-12 strain, a culture solution obtained by centrifugation of the fermentation culture or a dried product thereof, and the fermentation culture
  • the bacteria obtained by centrifugation or the dried product thereof are used for preparing veterinary drugs, feed additives and/or feeds for improving the smell of livestock and poultry; the odor of the feces of
  • livestock generally refers to pigs, sheep, cows, rabbits, chickens, quails, pigeons, ostriches, etc., which are within the knowledge of those skilled in the art of animal feed science.
  • animal feed science acceptable generally refers to the art of animal feed science, is not harmful to the product or to the animal, or has a reasonable or acceptable benefit/risk ratio.
  • auxiliary and excipient may be any conventional excipients and excipients in the field of animal feed science.
  • excipients or excipients that can be accepted as animal feed science include conventional excipients, excipients, diluents, fillers, solvents, support agents, premixes, disintegrants, surfactants in the field of animal feed science. And adsorption carriers and the like.
  • a flavoring agent, a sweetener, and the like may also be included as necessary.
  • Clostridium butyricum UCN-12 strain of the invention is applied in animal breeding, and can be produced as follows Beneficial effects:
  • the fermentation culture of the strain of the present invention, the fermentation broth or the bacterial body obtained by centrifugation of the fermentation culture, and the dried product of the fermentation broth or the bacterial body are used as active ingredients to feed the animal, and can effectively treat and/or prevent Clostridium welchii Disease, the effect is better than the existing antibiotics for the prevention and treatment of Clostridium perfringens;
  • the present invention generally and/or specifically describes the materials and test methods used in the test, and the examples are not particularly described in the examples.
  • the available dosages of the commercially available Clostridium butyricum products are all recommended dosages of the commercially available product specifications.
  • the main purpose of using existing commercially available Clostridium butyricum products is to compare the effects. While many of the materials and methods of operation used to accomplish the objectives of the present invention are well known in the art, the present invention is still described in detail herein. Field It will be apparent to the skilled person that in the following, the materials and methods of operation of the present invention are well known in the art unless otherwise stated.
  • Clostridium butyricum UCN-12 is deposited under the China Microbial Culture Collection Management Committee General Microbiology Center as CGMCC No.9652.
  • the new strain of Clostridium butyricum UCN-12 of the present invention is subjected to extraction of genomic DNA according to the procedure of Takala (MiniBEST Bacterial Genomic DNA Extraction Kit Ver. 3.0) kit, and 16S is amplified by Takala (16S rDNA Bacterial Identification PCR Kit) kit.
  • rDNA sequence Takara (MiniBEST DNA Fragment Purification Kit Ver.3.0 kit purification PCR product.
  • PCR amplification normal phase primer is 5'-GAGCGGATAACAATTTCACACAGG-3', reverse phase primer is 5'-CGCCAGGGTTTTCCCAGTCACGAC-3'.
  • Reaction system 50ul template DNA1ul; PCR Premix 25ul; Forward Premix 0.5ul; Reverse Premix 0.5ul; 16S-free 23ul; amplification conditions: 94 ° C preheating 5min; 94 ° C denaturation 1min; 55 ° C annealing 1min; 72 ° C extension 1.5min; 30 cycles; supplementation at 72 °C for 5 min; preservation at 4 ° C.
  • the amplified product was detected by 1% agarose gel electrophoresis, electrophoresis at 100 V/cm for 20 min, and photographed by Bio-Rad GelDoc image gel imaging system. Increasing the situation. It was found that there is only a single target band, no other bands that are not specific, and finally the PCR product is recovered. 2 000 bp P lus DNA marke is the standard molecular weight. Nucleic acid sequence is also referred .PCR product determined by TaKaRa Biotechnology Co., Ltd. (Dalian) batches by repeated measurements and calibration, Clostridium butyricum obtained UCN-12 of the present invention is a new strain 16S rDNA sequence as follows:
  • Clostridium butyricum UCN-12 strain was inoculated with 1-5% slope to inoculate Clostridium butyricum UCN-12 in the following fermentation medium: corn syrup dry powder 10g, ammonium sulfate 0.9g, magnesium sulfate 0.3g, sodium chloride 0.5 g, 30 g of glucose, 0.1 g of ferric chloride, 5 g of calcium carbonate, 1000 ml of water, adjusted to pH 6.2 to 7.4, sterilized by autoclaving at 121 ° C for 15 min, and cooled. The strain was cultured for 24 hours at 30-32 ° C for 24 hours.
  • the activated strain is inoculated with 10% of corn syrup dry powder, 0.9 g of ammonium sulfate, 0.3 g of magnesium sulfate, 0.5 g of sodium chloride, 30 g of glucose, 0.1 g of ferric chloride, 5 g of calcium carbonate, and 1000 ml of water, adjusted to pH 6.2. ⁇ 7.4, incubated in a liquid medium prepared by sterilizing at 121 ° C for 15 minutes in an autoclave, and incubated at 32 ° C for 24 hours as a seed liquid.
  • Clostridium butyricum UCN-12 strain was inoculated with 1-5% slope to inoculate Clostridium butyricum UCN-12 in the following fermentation medium: corn syrup dry powder 10g, ammonium sulfate 0.9g, magnesium sulfate 0.3g, sodium chloride 0.5g, glucose 30g, ferric chloride 0.1g, calcium carbonate 5g, water 1000ml, adjusted pH 6.2 ⁇ 7.4, sterilized in a pressure cooker at 121 ° C for 15min, cooled, 30-32 ° C fermentation culture 24 hours fermentation culture 24 hours activation Strain.
  • the activated strain is inoculated with 10 g of corn syrup dry powder, 0.9 g of ammonium sulfate, 0.3 g of magnesium sulfate, 0.5 g of sodium chloride, 30 g of glucose, 0.1 g of ferric chloride, 5 g of calcium carbonate, and 1000 ml of water.
  • the pH was adjusted to 6.2 to 7.4, and it was incubated in a liquid medium prepared by sterilizing at 121 ° C for 15 minutes in an autoclave, and cultured at 32 ° C for 24 hours as a seed liquid.
  • the cell suspension is collected from the cell suspension, 10 parts of the cells are taken, 30 parts of the auxiliary material are mixed, dried, pulverized, and packaged to obtain a solid powder composition, wherein the number of cells per 1 gram is quite small. At 3.3 ⁇ 10 8 .
  • Clostridium butyricum UCN-12 strain was inoculated with 1-5% slope to inoculate Clostridium butyricum UCN-12 in the following fermentation medium: corn syrup dry powder 10g, ammonium sulfate 0.9g, magnesium sulfate 0.3g, sodium chloride 0.5g, glucose 30g, ferric chloride 0.1g, calcium carbonate 5g, water 1000ml, adjusted pH 6.2 ⁇ 7.4, sterilized in a pressure cooker at 121 ° C for 15min, cooled, 30-32 ° C fermentation culture 24 hours fermentation culture 24 hours activation Strain.
  • the activated strain is inoculated with 10% of corn syrup dry powder, 0.9 g of ammonium sulfate, 0.3 g of magnesium sulfate, 0.5 g of sodium chloride, 30 g of glucose, 0.1 g of ferric chloride, 5 g of calcium carbonate, and 1000 ml of water, adjusted to pH 6.2. ⁇ 7.4, incubated in a liquid medium prepared by sterilizing at 121 ° C for 15 minutes in an autoclave, and incubated at 32 ° C for 24 hours as a seed liquid.
  • the cell suspension is collected by centrifuging the cell suspension, 10 parts of the cells are taken, 30 parts of the auxiliary material are mixed, dried, pulverized, granulated, and packaged to obtain a solid particle composition, wherein each 1 gram of the cells is obtained.
  • the number is not less than 3.3 ⁇ 10 8 .
  • Clostridium butyricum UCN-12 the other inhibitory effects of the fermented material of Clostridium butyricum UCN-12 of the present invention were determined (see Table 3), and the results show that the present invention is directed to the large intestine.
  • Bacillus, Salmonella, Vibrio anguillarum and Aeromonas hydrophila have better inhibitory effects and are superior to the existing commercially available Clostridium butyricum products. It is suggested that Clostridium butyricum UCN-12 can also be used for animal diarrhea and fish health.
  • the test selected chickens of similar weight and fed the fermentation broth obtained from the culture of Clostridium perfringens (Chicken source) purchased from the Chinese Veterinary Microorganisms Collection and Management Center until the typical Wei Symptoms of Clostridium disease, randomly grouped, 20 in each group, respectively in the same culture greenhouse.
  • the control group was fed a basal diet
  • the antibiotic group was fed with a basal diet supplemented with enramycin
  • the experimental group was fed a basic diet supplemented with 0.1%, 0.5%, 1.0%, 5.0%, and 10% Clostridium butyricum UCN-12. Free access to food and water, natural ventilation, natural light during the day, supplement lighting at night, for 3-5 consecutive days, observe the incidence (table 4).
  • the results showed that the experimental treatment group was significantly better than the basal diet group with antibiotics.
  • the existing commercially available Clostridium butyricum products had no therapeutic effect on avian Clostridium perfringens.
  • the experiment selected 200 white feather broilers with similar birth weights, which were randomly divided into 10 groups, 20 in each group, which were in the same culture greenhouse.
  • the control group was fed only with the basal diet, the antibiotic group was fed with the basal diet supplemented with enramycin, and the experimental group was supplemented with 0.1%, 0.5%, 1.0%, 5.0%, and 10% of Clostridium butyricum in the basal diet. .
  • the food was opened 24 hours after birth, and the fermentation culture of Clostridium butyricum UCN-12 was thoroughly mixed and fed. Free access to food and water, natural ventilation, natural daylight, supplemental lighting at night, vaccination as usual.
  • Clostridium perfringens (chicken source) produced by the Chinese veterinary microbial culture collection and management center was added to each group.
  • the test period was For 49 days, the incidence of Clostridium perfringens was observed (Table 5). The results showed that the experimental treatment group was significant Compared with the basic diet control group supplemented with antibiotics, the existing commercially available Clostridium butyricum products have no preventive effect on Clostridium perfringens.
  • the experiment selected 90 healthy pigs with similar weights of 70 days old, and fed the fermentation broth obtained from the culture of Clostridium perfringens (P. sphaeroides), which is purchased from the Chinese Veterinary Microorganisms Collection and Management Center. Until the typical symptoms of Clostridium perfringens, randomized groups, 10 in each group, were in the same culture greenhouse. The control group was fed the basal diet, the antibiotic group was fed the basal diet supplemented with enramycin, and the experimental group was fed with 0.5%, 1%, 2%, and 5.0% basal diet of Clostridium butyricum UCN-12. Food and water, natural ventilation, natural light during the day, for 3-5 days, observe the incidence (Table 6).
  • Clostridium perfringens P. sphaeroides
  • the test selected 90 healthy 70-day-old piglets, randomly divided into 6 groups, 10 in each group, respectively in the same breeding greenhouse.
  • the control group was fed the basal diet
  • the antibiotic group was fed with the basal diet supplemented with enramycin
  • the treatment group was fed with 0.5%, 1%, 2%, and 5.0% of Clostridium butyricum UCN-12 in the basal diet.
  • Clostridium perfringens Free access to food and water, natural ventilation, natural daylight, routine vaccination, continuous feeding for 10 days, adding 0.5% of the group's disease caused by Clostridium perfringens purchased from China Veterinary Microorganisms Collection and Management Center
  • the fermentation broth of Clostridium perfringens (porcine source) was cultured for 50 days, and the incidence of Clostridium perfringens was observed (Table 7). The results showed that the experimental treatment group was significantly better than the basal diet control group supplemented with antibiotics.
  • the existing commercially available Clostridium butyricum products had no preventive effect on Clostridium perfringens.
  • Blank control group 10 100 Antibiotic group 10 60 Clostridium butyricum UCN-12 fermentation culture 0.5% group 10 0 Clostridium butyricum UCN-12 fermentation culture 1.0% group 10 0 Clostridium butyricum UCN-12 fermentation culture 2.0% group 10 0 Clostridium butyricum UCN-12 fermentation culture 5.0% group 10 0 Commercially available Clostridium butyricum products 1 10 90 Commercially available Clostridium butyricum products 2 10 80 Commercially available Clostridium butyricum products 3 10 100
  • the experiment selected 90 healthy 90-day-old sheep, and fed the fermentation broth obtained from the culture of Clostridium perfringens (C. sinensis), which is purchased from the Chinese Veterinary Microorganisms Collection and Management Center. Until the typical symptoms of Clostridium perfringens, randomized groups, 10 in each group, were in the same culture greenhouse. The control group was fed the basal diet, the antibiotic group was fed the basal diet supplemented with enramycin, and the experimental group was fed with 0.5%, 1%, 2%, and 5.0% basal diet of Clostridium butyricum UCN-12. Food and water, natural ventilation, natural light during the day, for 3-5 consecutive days, observe the incidence (Table 8).
  • the experiment selected 90 healthy 90-day-old lambs, which were randomly divided into 9 groups, 10 in each group, which were in the same culture greenhouse.
  • the control group was fed the basal diet
  • the antibiotic group was fed with the basal diet supplemented with enramycin
  • the treatment group was fed with 0.5%, 1%, 2%, and 5.0% of Clostridium butyricum UCN-12 in the basal diet.
  • the culture medium of Clostridium perfringens (Yangyuan) was tested for 50 days and the incidence was observed (Table 9). The results showed that the experimental treatment group was significantly better than the basal diet control group supplemented with antibiotics.
  • the existing commercially available Clostridium butyricum products had no preventive effect on Clostridium perfringens.
  • the experiment selected 90 rabbits with similar weights of 90-day-old rabbits, and fed the fermentation broth obtained from the culture of Clostridium perfringens (Rabbit source) purchased from the Chinese Veterinary Microorganisms Collection and Management Center until the typical Symptoms of Clostridium perfringens were randomly divided into groups of 10 in each farm.
  • the control group was fed the basal diet
  • the antibiotic group was fed the basal diet supplemented with enramycin
  • the experimental group was fed with 0.5%, 1%, 2%, and 5.0% basal diet of Clostridium butyricum UCN-12.
  • the results showed that the experimental treatment group was significantly better than the basal diet group with antibiotics, and there was no non-growth rabbit in the basal diet group.
  • the existing commercially available Clostridium butyricum products had no treatment for rabbit Wisdom's disease. effect.
  • the experiment selected 90 rabbits with similar weights of 90-day-old rabbits, randomly divided into 9 groups, 10 in each group, respectively in the same culture greenhouse.
  • the control group was fed the basal diet
  • the antibiotic group was fed with the basal diet supplemented with enramycin
  • the treatment group was fed with 0.5%, 1%, 2%, and 5.0% of Clostridium butyricum UCN-12 in the basal diet.
  • Clostridium perfringens gas produced from the China Veterinary Microorganisms Collection and Management Center
  • the culture medium of Clostridium perfringens (rabbit source) was tested for 30 days to observe the occurrence of Clostridium perfringens (Table 11). The results showed that the experimental treatment group was significantly better than the basal diet control group supplemented with antibiotics.
  • the existing commercially available Clostridium butyricum products had no preventive effect on rabbit Clostridium perfringens.
  • the experiment selected 500 ducks with the same birth weight, and randomly divided into 10 groups, 50 in each group, which were in the same culture greenhouse.
  • the control group was only fed with the basic diet supplemented with antibiotics.
  • the experimental group was fed with the micro-ecological preparations of the composition in the basal diet according to the method of CN201010249225.2, and the other groups were added 0.1% and 0.5% respectively to the basal diet. 1.0%, 5.0%, and 10% of Clostridium butyricum UCN-12. Free access to food and water, natural ventilation, natural daylight, supplemental lighting at night, vaccination as usual.
  • the experiment selected 500 babies with similar birth weights on the 1st, and randomly divided into 10 groups, 50 in each group, which were respectively in the same culture greenhouse.
  • the control group was only fed with the basic diet supplemented with antibiotics.
  • the experimental group was fed with the micro-ecological preparations of the composition in the basal diet according to the method of CN201010249225.2, and the other groups were added 0.1% and 0.5% respectively to the basal diet. 1.0%, 5.0%, and 10% of Clostridium butyricum UCN-12. Free access to food and water, natural ventilation, natural daylight, supplemental lighting at night, vaccination as usual.
  • Clostridium butyricum UCN-12 has the taste of removing eggs, bright yellow egg color and the best taste.
  • the experiment selected 50 prenatal constipation sows, which were divided into control group and experimental group, with 10 heads in each group.
  • 50 postpartum constipation sows were selected and divided into control group and experimental group, with 10 heads in each group.
  • the control group was fed the basal diet.
  • 10 ml of the culture of Clostridium butyricum UCN-12 was added to the basal diet per day, and the mixture was mixed for 7 days. The treatment was observed (Table 18).
  • the existing commercially available Clostridium butyricum products have no therapeutic effect.
  • Control group 100 0.5% dose group 0 2% dose group 0 5% dose group 0
  • Clostridium butyricum products 1 postpartum constipation 90 Commercially available Clostridium butyricum products 2 postpartum constipation 80
  • Clostridium butyricum products 3 postpartum constipation 90 Commercially available Clostridium butyricum product 1 prenatal constipation 80
  • Clostridium butyricum products 2 prenatal constipation commercially available Clostridium butyricum products 3 prenatal constipation 80
  • a total of 900 white-feathered broilers of 1 day old with similar birth weights were selected and randomly divided into 9 groups of 100 animals each in the same culture greenhouse.
  • the control group was fed a basal diet supplemented with antibiotics, and the high, medium and low dose test groups were supplemented with 0.1%, 0.5%, 1.0%, 5.0%, and 10% of Clostridium butyricum UCN-12 in the basal diet. Before the start of the diet, all the groups were weighed, and the food was eaten 24 hours after birth.
  • the fermentation culture of Clostridium butyricum UCN-12 was thoroughly mixed and fed. Weigh once a week, freely eat and drink, naturally ventilate, natural light during the day, supplement lighting at night, and vaccinate as usual.
  • the test period was 49 days, and the weight gain and meat ratio were measured and calculated (Table 20). The results showed that the weight gain and meat-to-feed ratio of the experimental treatment group were significantly better than the basal diet control group supplemented with antibiotics, and also superior to the existing commercially available Clostridium butyricum products.
  • Seventy normal weaned piglets with similar weights were selected and randomly divided into 7 groups, 10 in each group, which were in the same culture greenhouse.
  • the control group was fed a basal diet supplemented with antibiotics, and the experimental group added 0.5%, 2%, and 5.0% of Clostridium butyricum UCN-12 to the basal diet. Free access to food and water, natural ventilation, natural daylight, vaccination as usual. Test The period was 180 days, and weight gain, meat to material ratio, and death were measured and calculated (Table 22). The results showed that the weight gain, meat-to-batch ratio and death number of the experimental treatment group were significantly better than the basic diet control group with antibiotic added, and also better than the existing commercially available Clostridium butyricum products.
  • Processing group Smelly score Remarks Control group 95 Extremely stinky 0.1% dose group 60 Slightly stinky 0.5% dose group 50 Slightly stinky
  • Seventy normal weaned piglets with similar weights were selected and randomly divided into 7 groups, 10 in each group, which were in the same culture greenhouse.
  • the control group was fed a basal diet supplemented with antibiotics, and the experimental group added 0.5%, 2%, and 5.0% of Clostridium butyricum UCN-12 to the basal diet. Free access to food and water, natural ventilation, natural daylight, vaccination as usual.
  • the test period was 180 days, and the human stool odor was judged by human scoring method (Table 24). The results showed that the experimental treatment was significantly better than the basal diet control group supplemented with antibiotics, and also superior to the existing commercially available Clostridium butyricum products.
  • Processing group Smelly score Remarks Control group 95 Extremely stinky 0.5% dose group 40 Slightly stinky 2% dose group 30 Slightly stinky 5% dose group 30 Slightly stinky Commercially available Clostridium butyricum products 1 85 Stinks Commercially available Clostridium butyricum products 2 80 Stinks Commercially available Clostridium butyricum products 3 80 Stinks
  • Seventy-eight sheep with a weight of nearly 90 days were selected and randomly divided into 7 groups, 70 in each group. They are in the same breeding greenhouse.
  • the control group was fed a basal diet supplemented with antibiotics, and the experimental group added 0.5%, 2%, and 5.0% of Clostridium butyricum UCN-12 to the basal diet. Free access to food and water, natural ventilation, natural daylight, vaccination as usual.
  • the test period was 180 days, and the human stool odor was judged by human scoring method (Table 25). The results showed that the experimental treatment was significantly better than the basal diet control group supplemented with antibiotics, and also superior to the existing commercially available Clostridium butyricum products.
  • Processing group Smelly score Remarks Control group 100 Extremely stinky 0.5% dose group 50 Slightly stinky 2% dose group 30 Slightly stinky 5% dose group 30 Slightly stinky Commercially available Clostridium butyricum products 1 80 Stinks Commercially available Clostridium butyricum products 2 85 Stinks Commercially available Clostridium butyricum products 3 80 Stinks
  • Clostridium butyricum UCN-12 has a protective effect on liver damage caused by undesirable chemicals in livestock and poultry, and can also eliminate endotoxin in animals, and is also suitable for regular health care of livestock and poultry.
  • the invention effectively overcomes the defects of the prior art, and is applied to 10 aquaculture enterprises in Chengdu and Chongqing, and has excellent effects on the treatment and/or prevention of Clostridium perfringens, and the mortality rate of livestock and poultry is reduced to zero.
  • the quality of livestock and poultry meat is all high quality.
  • the egg has a unique flavor and excellent quality.
  • the product is in short supply and the price almost doubles.
  • the new benefit of the farm has reached 20-30%, and the application demonstration has been very satisfactory, which is very popular among farmers.
  • the fermentation culture of the strain of the present invention the fermentation broth or the bacterial body obtained by centrifugation of the fermentation culture, and the dried product of the fermentation broth or the bacterial body are fed as active ingredients. It can treat and/or prevent Clostridium perfringens. It does not need to use antibiotics in the whole process, avoids the antibiotic residue affecting human health, completely eliminates the environmental pollution caused by the large use of antibiotics in livestock and poultry breeding, and can obviously raise the livestock.
  • the production performance and quality of poultry, reducing the stench of livestock and poultry, and achieving animal-friendly environment, the industrial application prospect of the present invention is very promising.

Abstract

提供了一种丁酸梭菌UCN-12菌株及其组合物和用途，该菌株的保藏编号为CGMCC No.9652。采用该菌株的发酵培养物、发酵培养物离心所得的发酵液或菌体、以及该发酵液或菌体的干燥物作为有效成分喂养畜禽，能治疗和/或预防畜禽魏氏梭菌病，能显著降低禽蛋中胆固醇和脂肪，去除禽蛋腥味，改善禽蛋风味，提高畜禽动物的生产性能，治疗和/或预防母猪产前产后便秘，以及减少畜禽便臭。

Description

丁酸梭菌UCN-12菌株及其组合物和应用 技术领域
本发明属于微生物学领域，涉及一种丁酸梭菌UCN-12菌株及其组合物和应用。
背景技术
魏氏梭菌病又称产气荚膜杆菌病，为食源性胃肠炎最常见病之一。国外统计魏氏梭菌引起的食物中毒占细菌性食物中毒的30％左右，最新统计数据表明，美国每年由产气荚膜梭菌引起的食物中毒人数在250,000左右，造成的经济损失达到1.2亿美元。流行病学调查发现，平均每起由产气荚膜梭菌引起的食物中毒事件的暴发，通常涉及50-100人。有些严重的食物中毒暴发甚至涉及到上千人。
魏氏梭菌一般可分为A、B、C、D、E和F六型，A型与E型菌及其产生的α为主要致病因素。该病原菌自消化道或伤口侵入机体，在小肠和盲肠绒毛膜上大量繁殖并产生强烈的α毒素，改变毛细血管的通透性，使毒素大量进入畜禽血液，引起全身性毒血症。
魏氏梭菌病也是畜禽常见疾病之一，每年都有大量的动物因不同类型魏氏梭菌感染而死亡。该病发病急、病程短、无任何前期症状而突然死亡，呈多地方性流行或散发，一年四季均可发生，尤以冬、春季为发病高峰期，严重时发病率达到100％,而且死亡率极高，达95％，是一种严重危害家兔生产的急性传染病。
现有技术中，防治魏氏梭菌病主要使用先锋霉素、氨苄青霉素、青霉素、氯霉素、庆大霉素和四环素等。由于长期使用抗生素造成的耐药性问题，致使抗生素使用量越来越高，治疗效果却越来越差，对环境污染也越来越大，特别药物残留安全更是越来越成为全社会担忧的问题。疫苗是一种较好地预防魏氏梭菌病的方法，但保护期短，且成本高，生产上难接受。
中国专利申请201210390182.9公开了及一种预防和治疗仔猪魏氏梭菌病的中兽药及其制备方法，所述配方由黄连10-20份、黄芩10-20份、连翘5-15份、金银花5-15份、当归10-20份、白芍10-20份、三七粉1-5份、人参茎叶5-15份和甘草5-10份组成，该方法难以消除人们对药物残留担忧，且成本高。专利申请201210552595.2公开了含地衣芽孢杆菌，枯草芽孢杆菌，屎肠球菌，粪链球菌、木聚糖酶的专用饲料，由于地衣芽孢杆菌和枯草芽孢杆菌与屎肠球菌和粪链球菌存在相互作用，组合物配伍欠合理，该方法效果也欠佳。专利申请201310577204.7和201210261894.0分别公开了一株枯草芽孢杆菌和一株耐酸、耐热、耐胆盐的地衣芽孢杆菌来防治魏氏梭菌病，所用菌种与本发明完全不同，同时，枯草芽孢杆菌和地衣芽孢杆菌属于好氧菌，需要通气培养，工艺复杂，生产成本也高于本发明。
本发明的发明人经过4年多的筛选、研究、试制，最终优选出活力强的丁酸梭菌UCN-12菌株。
发明内容
本发明所解决的第一个技术问题是提供了一种丁酸梭菌新菌株，采用本发明菌株的发酵培养物、发酵培养物离心所得的发酵液或菌体、以及该发酵液或菌体的干燥物为有效成分喂养动物，具有治疗和/或预防畜禽魏氏梭菌病，全过程无需添加抗生素，避免了抗生素残留影响人体健康，彻底解除动物养殖中因大量使用抗生素对环境的污染。还能显著降低禽蛋中胆固醇和脂肪，去除禽蛋腥味，改善鸡蛋风味，提高畜禽动物的生产性能，治疗和/或预防母猪产前产后便秘，减少畜禽便臭，实现养殖环境友好。
本发明新菌株为丁酸梭菌(Clostridium butyricum)，编号为UCN-12，于2014年9月15日保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，保藏编号为CGMCC №.9652。保藏地址： 北京市朝阳区北辰西路1号院3号中国科学院微生物研究所，邮编100101。于2016年3月10日由国内保藏转换为国际保藏。
丁酸梭菌UCN-12菌株的16S rDNA序列如SEQ ID No.1所示。
本发明丁酸梭菌UCN-12菌株与丁酸梭菌现有公开报道的菌株比较，除16s rDNA序列有区别外，在生化性状方面也有差异，见表1。最明显区别是丁酸梭菌UCN-12产己酸量是丁酸的10倍以上，有时可达到20倍，表现出特有己酸风味，而现有丁酸梭菌均不产己酸。
表1 丁酸梭菌UCN-12菌株与现有的丁酸梭菌菌株的生化性状比较
项目 丁酸梭菌UCN-12 丁酸梭菌* 丁酸梭菌** 丁酸梭菌*** 丁酸梭菌****
阿拉伯糖 + -+ + + +-
半乳糖 + + + + +
甘露糖 + + + + +
果糖 + + + + +
核糖 + + + - +
木糖 + + + + +
葡萄糖 + + + + +
山梨糖 - -+ - -  
鼠李糖 -+ - - - -
蜜二糖 -+ + + - +
麦芽糖 + + + + +
乳糖 -+ + + + +
纤维二糖 + + + + +
蔗糖 + + + + +
松二糖 + + + +  
淀粉 + + + -+ +
棉子糖 + + + - +
松三糖 - - - + -/+
菊糖 + -+ + + -/+
甘露醇 + - - -+ -/+
肌醇 - - - + -
山梨醇 - - - -+ -
卫矛醇 - - - -  
丙三醇 + + - - +
七叶苷 + + + + +
苦杏仁苷 - - - -+  
水杨素 + + + + +
明胶 - - - -  
产己酸 + - - -  
注：“+”试验结果为阳性，“-”试验结果为阴性，“-+”试验结果为弱阳性；“*”菌株分离于杭州惠嘉丰牧科技有限公司市售产品，“**”菌株分离于青岛东海药业有限公司市售产品酪酸菌片2，“****”菌株分离于中科生物工程有限公司市售产品；“****”数据来自《伯杰氏细菌鉴定手册》(第九版)；空格表示无数据。
本发明所解决的第二个技术问题是提供了以本发明丁酸梭菌UCN-12菌株为主要有效成分的组合物，可作为饲料添加剂用。所述的组合物含预防和/或治疗有效量的丁酸梭菌UCN-12菌株，丁酸梭菌UCN-12菌株的发酵培养物，该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物为有效成分，加入可接受的辅料制成。可接受的辅料为动物饲料科学可接受的载体或赋形 剂。所述组合物为单位剂量形式，优选的，其中所述的单位剂量形式选自：片剂、颗粒剂、粉剂、口服溶液、口服混悬液、口服乳剂和预混剂等。
本发明所解决的第三个技术问题是提供了丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于制备治疗或预防畜禽魏氏梭菌病的兽药、饲料添加剂和/或饲料中的应用。
本发明所解决的第四个技术问题是丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于制备降低禽蛋中胆固醇和脂肪，去除禽蛋腥味，改善禽蛋风味的兽药、饲料添加剂和/或饲料中的应用。发明人发现，不仅能够显著降低禽蛋中胆固醇和脂肪，还发现饲喂本发明丁酸梭菌UCN-12菌株及其衍生物后，可完全去除鸡蛋腥味，鸡蛋风味超过野生饲养土鸡蛋，且口感更好。本发明新菌株上述作用不仅提高了养殖动物产品健康品质，更重要是增加了产品市场附加值和生产效益。
本发明所解决的第五个技术问题是丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于提高畜禽的生产性能的兽药、饲料添加剂和/或饲料中的应用，实现动物养殖高产化。
本发明所解决的第六个技术问题是丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或 其干燥物，该发酵培养物离心所得的菌体或其干燥物用于制备治疗或预防母猪产前产后便秘的兽药、饲料添加剂和/或饲料中的应用。发明人发现，在母猪日粮中添加丁酸梭菌UCN-12菌株，可以杜绝母猪便秘发生。
本发明所解决的第七个技术问题是丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于制备改善畜禽便臭的兽药、饲料添加剂和/或饲料中的应用；使养殖动物粪便臭味显著减少，动物粪便对环境不良影响变小，实现了环境友好。
下面更进一步地描述本发明。
如本文使用的术语“畜禽”，通常是指猪、羊、牛、兔、鸡、鹌鹑、鸽子、鸵鸟等，这些都是动物饲料科学领域技术人员的知识范围内。
如本文使用的术语“动物饲料科学可接受的”通常是指动物饲料科学领域可使用，对产品或者对动物无害，或具有合理的或可接受的利益/风险比。
如本文使用的术语“辅料”和“赋形剂”可以是动物饲料科学领域中任何常规的辅料和赋形剂。具体的辅料和赋形剂的选择将取决于用于使用方式，用于特定动物养殖模式的合适饲料添加剂组合物的制备方法完全在动物饲料科学领域技术人员的知识范围内。例如，可以作为动物饲料科学可接受的辅料或赋形剂包括动物饲料科学领域常规的辅料、赋形剂、稀释剂、填充剂、溶剂、支持剂、预混剂、崩解剂、表面活性剂和吸附载体等。必要时，还可以包括香味剂和甜味剂等。
本发明丁酸梭菌UCN-12菌株在动物养殖方面应用，可产如下的 有益效果：
1.采用本发明菌株的发酵培养物、发酵培养物离心所得的发酵液或菌体、以及该发酵液或菌体的干燥物为有效成分喂养动物，能有效治疗和/或预防魏氏梭菌病，效果优于现有防治魏氏梭菌病专用抗生素恩拉霉素；
2.全过程无需添加抗生素，避免了抗生素残留影响人体健康，彻底解除动物养殖中因大量使用抗生素对环境的污染；
3.能显著降低禽蛋中胆固醇和脂肪含量，去除腥味，改善饲养鸡蛋风味，达到饲养鸡蛋风味与野生饲养土鸡蛋完全一样，提高禽禽蛋产品品质和附加值；
4.减少畜禽类动物便臭，实现养殖环境友好；
5.可显著促进动物健康生长，提高生产性能，并降低成本；
6.能有效治疗和/或预防母猪产前产后便秘。
具体实施方式
为表明本发明丁酸梭菌UCN-12对动物健保的效果，下面通过具体的实施例或试验例进一步说明本发明，但是，应当理解为，这些实施例或试验例仅仅是用于更详细具体地说明之用，而不应理解为用于以任何形式限制本发明。
本发明对试验中所使用到的材料以及试验方法进行一般性和/或具体的描述，实施例中未特别说明，现有市售丁酸梭菌产品使用剂量均为该市售产品说明书推荐剂量，使用现有市售丁酸梭菌产品主要目的比较效果差异。虽然为实现本发明目的所使用的许多材料和操作方法是本领域公知的，但是本发明仍然在此作尽可能详细描述。本领域 技术人员清楚，在下文中，如果未特别说明，本发明所用材料和操作方法是本领域公知的。
丁酸梭菌UCN-12新菌株的16S rDNA序列测定实施例
丁酸梭菌(Clostridium butyricum)UCN-12，在中国微生物菌种保藏管理委员会普通微生物中心的保藏编号为CGMCC No.9652。本发明的丁酸梭菌UCN-12新菌株，按照Takala(MiniBEST Bacterial Genomic DNA Extraction Kit Ver.3.0)试剂盒操作步骤，经过提取基因组DNA，Takala(16S rDNA Bacterial Identification PCR Kit)试剂盒扩增16S rDNA序列,Takara(MiniBEST DNA Fragment Purification Kit Ver.3.0试剂盒纯化PCR产物。PCR扩增：正相引物为5'-GAGCGGATAACAATTTCACACAGG-3'，反相引物为5'-CGCCAGGGTTTTCCCAGTCACGAC-3'。反应体系50ul：模板DNA1ul；PCR Premix 25ul；Forward Premix 0.5ul；Reverse Premix 0.5ul；16S-free 23ul；扩增条件：94℃预热性5min；94℃变性1min；55℃退火1min；72℃延伸1.5min；30个循环；72℃补充延伸5min；4℃保存。扩增产物经1％琼脂糖凝胶电泳检测，100V/cm下稳压电泳20min，Bio-Rad GelDoc image凝胶成像系统照相，观察其扩增情况。发现只有单一的目标带，没有非特异性的其他条带，最后回收PCR产物。2 000bp P lus DNA marke为标准分子量标记。PCR产物的核酸序列由宝生物工程(大连)有限公司测定。通过多批次重复测定和校准，获得本发明的丁酸梭菌UCN-12新菌株16S rDNA序列如下：
Figure PCTCN2016078912-appb-000001
Figure PCTCN2016078912-appb-000002
制备实施例1发酵培养物制备
取丁酸梭菌UCN-12菌株，按1-5％斜面接种丁酸梭菌UCN-12于下述发酵培养基中：玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml，调节pH6.2～7.4，用高压锅在121℃灭菌15min，冷却。30-32℃发酵培养24小时发酵培养24小时活化菌株。
活化菌株按1-5％接种于玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml，调节pH6.2～7.4，用高压锅在121℃灭菌15min冷却后制成的液体培养基中，在32℃下培养24小时，作为种子液。
按1-5％接种于玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml，调节pH6.2～7.4，用高压锅在121℃灭菌15min冷却后制成的液体培养基中，在30℃下发酵罐发酵培养24小时，获发酵培养物，其中每1ml菌细胞数量不少于108。
制备实施例2发酵培养物组合物制备
取丁酸梭菌UCN-12菌株，按1-5％斜面接种丁酸梭菌UCN-12于下述发酵培养基中：玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml，调节pH6.2～7.4，用高压锅在121℃灭菌15min，冷却,30-32℃发酵培养24小时发酵培养24小时活化菌株。
活化菌株按1-5％接种于玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml， 调节pH6.2～7.4，用高压锅在121℃灭菌15min冷却后制成的液体培养基中，在32℃下培养24小时，作为种子液。
按1-5％接种于玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml，调节pH6.2～7.4，用高压锅在121℃灭菌15min冷却后制成的液体培养基中，在32℃下发酵罐发酵培养24小时，获发酵培养物，其中每1ml菌细胞数量不少于108。
在无菌条件下，离心菌细胞悬液收集菌细胞，取菌细胞10份，加辅料30份混合，干燥，粉碎，分装，即得固体粉末组合物，其中每1克菌细胞数量不少于3.3×108。
制备实施例3发酵培养物组合物制备
取丁酸梭菌UCN-12菌株，按1-5％斜面接种丁酸梭菌UCN-12于下述发酵培养基中：玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml，调节pH6.2～7.4，用高压锅在121℃灭菌15min，冷却，30-32℃发酵培养24小时发酵培养24小时活化菌株。
活化菌株按1-5％接种于玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml，调节pH6.2～7.4，用高压锅在121℃灭菌15min冷却后制成的液体培养基中，在32℃下培养24小时，作为种子液。
按1-5％接种于玉米浆干粉10g、硫酸铵0.9g、硫酸镁0.3g、氯化钠0.5g、葡萄糖30g、氯化铁0.1g、碳酸钙5g，水1000ml，调节pH6.2～7.4，用高压锅在121℃灭菌15min冷却后制成的液体培养基 中，在32℃下发酵罐发酵培养24小时，获发酵培养物，其中每1ml菌细胞数量不少于108。
在无菌条件下，离心菌细胞悬液收集菌细胞，取菌细胞10份，加辅料30份混合，干燥，粉碎，制粒、分装，即得固体颗粒组合物，其中每1克菌细胞数量不少于3.3×108。
生物学实施例1 丁酸梭菌UCN-12发酵物对魏氏梭菌病致病菌产 权荚膜梭菌的抑制作用
按照文献《微生物学通报》，2007年34卷4期：753-756方法，测定了本发明丁酸梭菌UCN-12发酵物对不同来源的魏氏梭菌病致病菌—产气荚膜梭菌的抑制作用(见表2)，结果显示本发明对产气荚膜梭菌有强烈抑制作用，优于恩拉霉素，现有市售丁酸梭菌产品对魏氏梭菌病致病菌产气荚膜梭菌无抑制作用。
表2 丁酸梭菌UCN-12及其组合物的抗魏氏梭菌病致病菌作用(抑菌圈直径，mm)
Figure PCTCN2016078912-appb-000003
注*-：示无抑制作用
生物学实施例2 丁酸梭菌UCN-12发酵物对其它致病菌的抑制作 用
按照文献《微生物学通报》，2007年34卷4期：753-756方法，测定了本发明丁酸梭菌UCN-12发酵物对其它的抑制作用(见表3)，结果显示本发明对大肠杆菌、沙门氏杆菌、鳗弧菌和嗜水气单胞菌有较好抑制作用，优于现有市售丁酸梭菌产品。提示丁酸梭菌UCN-12也可用于动物腹泻和鱼类保健。
表3 丁酸梭菌UCN-12及其组合物的抑菌作用(抑菌圈直径，mm)
编号 类别 大肠杆菌 沙门氏杆菌 鳗弧菌 嗜水气单胞菌
1 丁酸梭菌UCN-12发酵物 35 37 39 35
2 丁酸梭菌UCN-12发酵液 39 29 36 31
3 丁酸梭菌UCN-12粉末 34 36 32 27
4 市售丁酸梭菌产品1 18 23 21 -
5 市售丁酸梭菌产品2 - 24 - 15
6 市售丁酸梭菌产品3 17 22 - 12
注*-：示无抑制作用
应用实施例1 治疗禽类魏氏梭菌病作用
试验选择重量相近健康鸡，分别喂食从中国兽医微生物菌种保藏管理中心购置的魏氏梭菌病致病菌产气荚膜梭菌(鸡源)经过培养获得的发酵培养液，直至出现典型魏氏梭菌病症状，随机分组，每组20只，分别于同一养殖大棚。对照组喂基础日粮，抗生素组喂添加恩拉霉素的基础日粮，试验组分别喂添加0.1％、0.5％、1.0％、5.0％和10％丁酸梭菌UCN-12的基础日粮，自由采食和饮水，自然通风，白天自然光照，晚间补充灯光照明，连续3-5天，观察发病情况(表 4)。结果显示实验处理组明显优于添加抗生素的基础日粮组，现有市售丁酸梭菌产品对禽类魏氏梭菌病无治疗作用。
表4 丁酸梭菌UCN-12发酵培养物对禽类魏氏梭菌病的治疗作用
处理组 动物数(只) 治愈率(％)
空白对照组 20 0
抗生素组 20 50
丁酸梭菌UCN-12发酵培养物0.1％组 20 100
丁酸梭菌UCN-12发酵培养物0.5％组 20 100
丁酸梭菌UCN-12发酵培养物1.0％组 20 100
丁酸梭菌UCN-12发酵培养物5.0％组 20 100
丁酸梭菌UCN-12发酵培养物10.0％组 20 100
市售丁酸梭菌产品1 20 0
市售丁酸梭菌产品2 20 0
市售丁酸梭菌产品3 20 0
应用实施例2 预防禽类魏氏梭菌病作用
试验选择出生重量相近的1日白羽肉鸡200只，随机分成10组，每组20只，分别于同一养殖大棚。对照组只饲喂基础日粮，抗生素组喂添加恩拉霉素的基础日粮，试验组分别在基础日粮中添加0.1％、0.5％、1.0％、5.0％和10％的丁酸梭菌。出生后24h开食，将丁酸梭菌UCN-12发酵培养物充分混匀后饲喂。自由采食和饮水，自然通风，白天自然光照，晚间补充灯光照明，按常规接种疫苗。连续饲喂10天后，在各组中添加0.5％的从中国兽医微生物菌种保藏管理中心购置的魏氏梭菌病致病菌产气荚膜梭菌(鸡源)发酵培养液，试验期为49天，观察魏氏梭菌病发生情况(表5)。结果显示实验处理组显著 优于添加抗生素的基础日粮对照组，现有市售丁酸梭菌产品对鸡魏氏梭菌病无预防作用。
表5 丁酸梭菌UCN-12发酵培养物对鸡魏氏梭菌病预防作用
处理组 动物数(只) 发病率(％)
空白对照组 20 80
抗生素组 20 60
丁酸梭菌UCN-12发酵培养物0.1％组 20 0
丁酸梭菌UCN-12发酵培养物0.5％组 20 0
丁酸梭菌UCN-12发酵培养物1.0％组 20 0
丁酸梭菌UCN-12发酵培养物5.0％组 20 0
丁酸梭菌UCN-12发酵培养物10.0％组 20 0
市售丁酸梭菌产品1 20 90
市售丁酸梭菌产品2 20 80
市售丁酸梭菌产品3 20 90
应用实施例3 治疗猪魏氏梭菌病作用
试验选择重量相近健康70天龄仔猪90只，喂食从中国兽医微生物菌种保藏管理中心购置的魏氏梭菌病致病菌产气荚膜梭菌(猪源)经过培养获得的发酵培养液，直至出现典型魏氏梭菌病症状，随机分组，每组10只，分别于同一养殖大棚。对照组喂基础日粮，抗生素组喂添加恩拉霉素的基础日粮，试验组分别喂添加0.5％、1％、2％和5.0％丁酸梭菌UCN-12的基础日粮，自由采食和饮水，自然通风，白天自然光照，连续3-5天，观察发病情况(表6)。结果显示实验处理组明显优于添加抗生素的基础日粮组，且未见抗生素的基础日粮组出现不生长的僵猪，现有市售丁酸梭菌产品对猪魏氏梭菌病无治疗作 用。
表6 丁酸梭菌UCN-12发酵培养物对猪魏氏梭菌病治疗作用
处理组 动物数(只) 治愈率(％)
空白对照组 10 0
抗生素组 10 40
丁酸梭菌UCN-12发酵培养物0.5％组 10 100
丁酸梭菌UCN-12发酵培养物1％组 10 100
丁酸梭菌UCN-12发酵培养物2％组 10 100
丁酸梭菌UCN-12发酵培养物5％组 10 100
市售丁酸梭菌产品1 10 0
市售丁酸梭菌产品2 10 0
市售丁酸梭菌产品3 10 0
应用实施例4 预防猪魏氏梭菌病作用
试验选择重量相近健康70天龄仔猪90只，随机分成6组，每组10只，分别于同一养殖大棚。对照组喂基础日粮，抗生素组喂添加恩拉霉素的基础日粮，处理组喂分别在基础日粮中添加0.5％、1％、2％和5.0％的丁酸梭菌UCN-12，自由采食和饮水，自然通风，白天自然光照，按常规接种疫苗，连续饲喂10天后，在各组中添加0.5％的从中国兽医微生物菌种保藏管理中心购置的魏氏梭菌病致病菌产气荚膜梭菌(猪源)发酵培养液，试验期为50天，观察魏氏梭菌病发生情况(表7)。结果显示实验处理组明显优于添加抗生素的基础日粮对照组，现有市售丁酸梭菌产品对猪魏氏梭菌病无预防作用。
表7 丁酸梭菌UCN-12发酵培养物对猪魏氏梭菌病的预防作用
处理组 动物数(只) 发病率(％)
空白对照组 10 100
抗生素组 10 60
丁酸梭菌UCN-12发酵培养物0.5％组 10 0
丁酸梭菌UCN-12发酵培养物1.0％组 10 0
丁酸梭菌UCN-12发酵培养物2.0％组 10 0
丁酸梭菌UCN-12发酵培养物5.0％组 10 0
市售丁酸梭菌产品1 10 90
市售丁酸梭菌产品2 10 80
市售丁酸梭菌产品3 10 100
应用实施例5 治疗羊魏氏梭菌病作用
试验选择重量相近健康90日龄羊90只，喂食从中国兽医微生物菌种保藏管理中心购置的魏氏梭菌病致病菌产气荚膜梭菌(羊源)经过培养获得的发酵培养液，直至出现典型魏氏梭菌病症状，随机分组，每组10只，分别于同一养殖大棚。对照组喂基础日粮，抗生素组喂添加恩拉霉素的基础日粮，试验组分别喂添加0.5％、1％、2％和5.0％丁酸梭菌UCN-12的基础日粮，自由采食和饮水，自然通风，白天自然光照，连续3-5天，观察发病情况(表8)。结果显示实验处理组明显优于添加抗生素的基础日粮组，且未见抗生素的基础日粮组出现不生长的僵羊，现有市售丁酸梭菌产品对羊魏氏梭菌病无治疗作用。
表8 丁酸梭菌UCN-12发酵培养物对羊魏氏梭菌病治疗作用
处理组 动物数(只) 治愈率(％)
空白对照组 10 0
抗生素组 10 40
丁酸梭菌UCN-12发酵培养物0.5％组 10 100
丁酸梭菌UCN-12发酵培养物1％组 10 100
丁酸梭菌UCN-12发酵培养物2％组 10 100
丁酸梭菌UCN-12发酵培养物5％组 10 100
市售丁酸梭菌产品1 10 0
市售丁酸梭菌产品2 10 0
市售丁酸梭菌产品3 10 0
应用实施例6 预防羊魏氏梭菌病作用
试验选择重量相近健康90日龄仔羊90只，随机分成9组，每组10只，分别于同一养殖大棚。对照组喂基础日粮，抗生素组喂添加恩拉霉素的基础日粮，处理组喂分别在基础日粮中添加0.5％、1％、2％和5.0％的丁酸梭菌UCN-12，自由采食和饮水，自然通风，白天自然光照，按常规接种疫苗，连续饲喂10天后，在各组中添加0.5％的从中国兽医微生物菌种保藏管理中心购置的羊魏氏梭菌病产气荚膜梭菌(羊源)发酵培养液，试验期为50天，观察发病情况(表9)。结果显示实验处理组明显优于添加抗生素的基础日粮对照组，现有市售丁酸梭菌产品对羊魏氏梭菌病无预防作用。
表9 丁酸梭菌UCN-12发酵培养物对羊魏氏梭菌病的预防作用
处理组 动物数(只) 发病率(％)
空白对照组 10 100
抗生素组 10 60
丁酸梭菌UCN-12发酵培养物0.5％组 10 0
丁酸梭菌UCN-12发酵培养物1.0％组 10 0
丁酸梭菌UCN-12发酵培养物2.0％组 10 0
丁酸梭菌UCN-12发酵培养物5.0％组 10 0
市售丁酸梭菌产品1 10 100
市售丁酸梭菌产品2 10 100
市售丁酸梭菌产品3 10 90
应用实施例7 治疗兔魏氏梭菌病作用
试验选择重量相近健康90日龄兔90只，喂食从中国兽医微生物菌种保藏管理中心购置的魏氏梭菌病产气荚膜梭菌(兔源)经过培养获得的发酵培养液，直至出现典型魏氏梭菌病症状，随机分组，每组10只，分别于同一养殖大棚。对照组喂基础日粮，抗生素组喂添加恩拉霉素的基础日粮，试验组分别喂添加0.5％、1％、2％和5.0％丁酸梭菌UCN-12的基础日粮，自由采食和饮水，自然通风，白天自然光照，连续3-5天，观察发病情况(表10)。结果显示实验处理组明显优于添加抗生素的基础日粮组，且未见抗生素的基础日粮组出现不生长的僵兔，现有市售丁酸梭菌产品对兔魏氏梭菌病无治疗作用。
表10 丁酸梭菌UCN-12发酵培养物对兔魏氏梭菌病治疗作用
处理组 动物数(只) 治愈率(％)
空白对照组 10 0
抗生素组 10 40
丁酸梭菌UCN-12发酵培养物0.5％组 10 80
丁酸梭菌UCN-12发酵培养物1％组 10 100
丁酸梭菌UCN-12发酵培养物2％组 10 100
丁酸梭菌UCN-12发酵培养物5％组 10 100
市售丁酸梭菌产品1 10 0
市售丁酸梭菌产品2 10 0
市售丁酸梭菌产品3 10 0
应用实施例8 预防兔魏氏梭菌病作用
试验选择重量相近健康90日龄兔90只，随机分成9组，每组10只，分别于同一养殖大棚。对照组喂基础日粮，抗生素组喂添加恩拉霉素的基础日粮，处理组喂分别在基础日粮中添加0.5％、1％、2％和5.0％的丁酸梭菌UCN-12，自由采食和饮水，自然通风，白天自然光照，按常规接种疫苗，连续饲喂10天后，在各组中添加0.5％的从中国兽医微生物菌种保藏管理中心购置的魏氏梭菌病产气荚膜梭菌(兔源)发酵培养液，试验期为30天，观察魏氏梭菌病发生情况(表11)。结果显示实验处理组明显优于添加抗生素的基础日粮对照组，现有市售丁酸梭菌产品对兔魏氏梭菌病无预防作用。
表11 丁酸梭菌UCN-12发酵培养物对兔魏氏梭菌病的预防作用
处理组 动物数(只) 发病率(％)
空白对照组 10 100
抗生素组 10 60
丁酸梭菌UCN-12发酵培养物0.5％组 10 30
丁酸梭菌UCN-12发酵培养物1.0％组 10 0
丁酸梭菌UCN-12发酵培养物2.0％组 10 0
丁酸梭菌UCN-12发酵培养物5.0％组 10 0
市售丁酸梭菌产品1 10 100
市售丁酸梭菌产品2 10 90
市售丁酸梭菌产品3 10 100
应用实施例9 降低鸡蛋中胆固醇、脂肪和改善风味作用
试验选择出生重量相近的1日龄罗曼褐壳蛋鸡1000只，随机分成10组，每组100只，分别于同一养殖大棚。对照组只饲喂添加抗生素的基础日粮，试验组一组按CN201010249225.2方法在基础日粮中添加组合物微生态制剂喂饲，其余组分别在基础日粮中添加0.1％、0.5％、1.0％、5.0％和10％的丁酸梭菌UCN-12。自由采食和饮水，自然通风，白天自然光照，晚间补充灯光照明，按常规接种疫苗。产蛋后试验组各随机抽取700g鸡蛋，按照GB/T 2220-2008《食品中胆固醇测定高效液相色谱法》和GB T 5009.6-2003食品中脂肪的测定进行测定(表12)。结果显示实验处理组胆固醇和脂肪含量显著低于添加抗生素的基础日粮对照组和现有技术方法CN201010249225.2组。提示丁酸梭菌UCN-12发酵培养物能降低鸡蛋中胆固醇和脂肪含量，现有技术方法CN201010249225.2对降低鸡蛋中胆固醇和脂肪含量无效，现有市售丁酸梭菌产品对降低鸡蛋中胆固醇和脂肪含量无作用。
表12 丁酸梭菌UCN-12发酵培养物对罗曼褐壳鸡蛋中胆固醇和脂肪的影响
处理组 胆固醇(X±s)/mg/100g 脂肪(X±s)/mg/100g
市场销售的有机鸡蛋 410.11±3.25 10.61±2.34
对照组 466.04±2.56 8.82±1.78
CN201010249225.2方法组 477.23±3.47 8.69±2.97
0.1％剂量组 282.48±1.84 6.12±1.31
0.5％剂量组 289.13±1.12 5.02±2.11
1％剂量组 248.46±2.34 4.83±1.43
5％剂量组 177.32±2.12 4.76±1.36
10％剂量组 183.65±1.83 5.24±1.51
市售丁酸梭菌产品1 419.23±1.62 11.23±1.79
市售丁酸梭菌产品2 435.71±1.59 12.34±1.83
市售丁酸梭菌产品3 410.11±1.93 10.58±1.71
鸡蛋风味采取人闻、品尝、观察打分方法评判，试验结果见表12，丁酸梭菌UCN-12具有去除鸡蛋腥味、蛋黄色泽鲜亮和最佳的口感。
表13 丁酸梭菌UCN-12发酵培养物对罗曼褐壳鸡蛋风味的影响
处理组 腥味 蛋黄色泽喜欢度 口感
市场销售的有机鸡蛋 + 80 70
土鸡蛋对照组 + 80 80
CN201010249225.2方法组 + 70 60
0.1％剂量组 - 100 95
0.5％剂量组 - 100 100
1％剂量组 - 100 100
5％剂量组 - 100 100
10％剂量组 - 100 100
市售丁酸梭菌产品1 + 80 70
市售丁酸梭菌产品2 + 70 60
市售丁酸梭菌产品3 + 70 60
注：+代表有腥味，-代表无腥味。
应用实施例10 降低鸭蛋中胆固醇、脂肪和改善风味作用
试验选择出生重量相近的1日鸭500只，随机分成10组，每组50只，分别于同一养殖大棚。对照组只饲喂添加抗生素的基础日粮，试验组一组按CN201010249225.2方法在基础日粮中添加组合物微生态制剂喂饲，其余组分别在基础日粮中添加0.1％、0.5％、1.0％、5.0％和10％的丁酸梭菌UCN-12。自由采食和饮水，自然通风，白天自然光照，晚间补充灯光照明，按常规接种疫苗。产蛋后试验组各随机抽取700g鸡蛋，按照GB/T 2220-2008《食品中胆固醇测定高效液相色谱 法》和GB T 5009.6-2003食品中脂肪的测定进行测定(表14)。结果显示实验处理组胆固醇和脂肪含量显著低于添加抗生素的基础日粮对照组和现有技术方法CN201010249225.2组。提示丁酸梭菌UCN-12发酵培养物能降低鸭蛋中胆固醇和脂肪含量，而现有技术方法CN201010249225.2对降低鸭蛋中胆固醇和脂肪含量无效，现有市售丁酸梭菌产品对降低鸭蛋中胆固醇和脂肪含量无作用。
表14 丁酸梭菌UCN-12发酵培养物对鸭蛋中胆固醇和脂肪的影响
处理组 胆固醇(X±s)/mg/100g 脂肪(X±s)/mg/100g
市场销售的有机鸡蛋 612.11±2.11 18.69±1.43
对照组 666.04±2.16 16.82±1.28
CN201010249225.2方法组 687.13±1.49 17.63±1.67
0.1％剂量组 352.22±1.06 8.12±1.35
0.5％剂量组 348.12±1.09 7.02±1.11
1％剂量组 332.46±2.01 7.33±1.27
5％剂量组 330.32±1.12 7.24±1.76
10％剂量组 353.15±1.43 7.43±1.46
市售丁酸梭菌产品1 615.36±1.26 17.21±1.73
市售丁酸梭菌产品2 608.41±1.48 15.23±1.81
市售丁酸梭菌产品3 632.15±1.51 16.97±1.47
鸭蛋风味采取人闻、品尝、观察打分方法评判，试验结果见表15，丁酸梭菌UCN-12具有去除鸡蛋腥味、蛋黄色泽鲜亮和最佳的口感。
表15 丁酸梭菌UCN-12发酵培养物对鸭蛋风味的影响
处理组 腥味 蛋黄色泽喜欢度 口感
市场销售的有机鸭蛋 + 60 70
对照组 + 60 50
CN201010249225.2方法组 + 60 60
0.1％剂量组 - 100 100
0.5％剂量组 - 100 100
1％剂量组 - 100 100
5％剂量组 - 100 100
10％剂量组 - 100 100
市售丁酸梭菌产品1 + 70 60
市售丁酸梭菌产品2 + 60 60
市售丁酸梭菌产品3 + 60 60
注：+代表有腥味，-代表无腥味。
应用实施例11 降低鹌鹑蛋中胆固醇、脂肪和改善风味作用
试验选择出生重量相近的1日鹌鹑500只，随机分成10组，每组50只，分别于同一养殖大棚。对照组只饲喂添加抗生素的基础日粮，试验组一组按CN201010249225.2方法在基础日粮中添加组合物微生态制剂喂饲，其余组分别在基础日粮中添加0.1％、0.5％、1.0％、5.0％和10％的丁酸梭菌UCN-12。自由采食和饮水，自然通风，白天自然光照，晚间补充灯光照明，按常规接种疫苗。产蛋后试验组各随机抽取700g鸡蛋，按照GB/T 2220-2008《食品中胆固醇测定高效液相色谱法》和GB T 5009.6-2003食品中脂肪的测定进行测定(表16)。结果显示实验处理组胆固醇和脂肪含量显著低于添加抗生素的基础日粮对照组和现有技术方法CN201010249225.2组。提示丁酸梭菌UCN-12发酵培养物能降低鹌鹑蛋中胆固醇和脂肪含量，而现有技术方法CN201010249225.2对降低鹌鹑蛋中胆固醇和脂肪含量无效，现有市售丁酸梭菌产品对降低鹌鹑蛋中胆固醇和脂肪含量无作用。
表16 丁酸梭菌UCN-12发酵培养物对鹌鹑蛋中胆固醇和脂肪的影响
处理组 胆固醇(X±s)/mg/100g 脂肪(X±s)/mg/100g
市场销售的有机鸭蛋 512.01±1.11 11.79±1.83
对照组 532.14±1.17 10.81±1.45
CN201010249225.2方法组 557.13±1.51 10.63±2.42
0.1％剂量组 362.29±1.56 7.15±1.79
0.5％剂量组 358.10±1.73 7.52±1.48
1％剂量组 352.83±1.69 6.53±1.57
5％剂量组 340.02±1.42 7.04±1.47
10％剂量组 359.14±1.78 7.23±1.66
市售丁酸梭菌产品1 522.22±1.41 10.73±1.97
市售丁酸梭菌产品2 509.43±1.32 10.07±1.38
市售丁酸梭菌产品3 529.13±1.76 9.08±1.29
鹌鹑蛋风味采取人闻、品尝、观察打分方法评判，试验结果见表16，丁酸梭菌UCN-12具有去除鸡蛋腥味、蛋黄色泽鲜亮和最佳的口感。
表17 丁酸梭菌UCN-12发酵培养物对鹌鹑蛋风味的影响
处理组 腥味 蛋黄色泽喜欢度 口感
市场销售的有机鹌鹑蛋 + 60 70
土鹌鹑蛋对照组 + 60 60
CN201010249225.2方法组 + 70 60
0.1％剂量组 - 100 100
0.5％剂量组 - 100 100
1％剂量组 - 100 100
5％剂量组 - 100 100
10％剂量组 - 100 100
市售丁酸梭菌产品1 + 70 60
市售丁酸梭菌产品2 + 70 60
市售丁酸梭菌产品3 + 70 60
注：+代表有腥味，-代表无腥味。
应用实施例12 治疗母猪产前产后便秘
试验选择产前便秘母猪50头，分为对照组和试验组，每组10头。 选择产后便秘母猪各50头，分为对照组和试验组，每组10头。对照组饲喂基础日粮，试验组按每头母猪每天在基础日粮中添加丁酸梭菌UCN-12发酵培养物10ml，拌匀后连续饲喂7天，观察治疗情况(表18)，现有市售丁酸梭菌产品无治疗作用。
表18 丁酸梭菌UCN-12发酵培养物对母猪便秘的治疗作用
处理组 剂量(ml/头) 治愈率(％)
产前便秘对照组 0 0
产后便秘对照组 0 0
产前便秘治疗组 10 100
产后便秘治疗组 10 100
市售丁酸梭菌产品1产后便秘 20 0
市售丁酸梭菌产品2产后便秘 20 0
市售丁酸梭菌产品3产后便秘 20 0
市售丁酸梭菌产品1产前便秘 20 0
市售丁酸梭菌产品2产前便秘 20 0
市售丁酸梭菌产品3产前便秘 20 0
应用实施例13 预防母猪产前产后便秘
母猪100只，随机分成10组，每组10只，分别于同一养殖大棚。对照组只饲喂添加抗生素的基础日粮，试验组分别在基础日粮中添加0.5％、2％和5.0％的丁酸梭菌UCN-12。自由采食和饮水，自然通风，白天自然光照，按常规接种疫苗。试验期为180天，观察便秘情况(表19)。结果显示实验处理明显优于添加抗生素的基础日粮对照组，现有市售丁酸梭菌产品无显著预防作用。
表19 丁酸梭菌UCN-12发酵培养物对母猪便秘的预防作用
处理组 母猪便秘发生率(％)
对照组 100
0.5％剂量组 0
2％剂量组 0
5％剂量组 0
市售丁酸梭菌产品1产后便秘 90
市售丁酸梭菌产品2产后便秘 80
市售丁酸梭菌产品3产后便秘 90
市售丁酸梭菌产品1产前便秘 80
市售丁酸梭菌产品2产前便秘 80
市售丁酸梭菌产品3产前便秘 80
应用实施例14 提高禽类养殖动物生产性能
试验选择出生重量相近的1日龄白羽肉鸡900只，随机分成9组，每组100只，分别于同一养殖大棚。对照组饲喂添加抗生素的基础日粮，高中低剂量试验组分别在基础日粮中添加0.1％、0.5％、1.0％、5.0％和10％的丁酸梭菌UCN-12。在开食前各组全部称出出生重量，出生后24h开食，将丁酸梭菌UCN-12发酵培养物充分混匀后饲喂。每周称重1次，自由采食和饮水，自然通风，白天自然光照，晚间补充灯光照明，按常规接种疫苗。试验期为49天，测定并计算增重和肉料比情况(表20)。结果显示实验处理组增重和肉料比明显优于添加抗生素的基础日粮对照组，也优于现有市售丁酸梭菌产品。
表20 丁酸梭菌UCN-12发酵培养物对白羽肉鸡生长性能影响
处理组 增重(kg) 肉料比
对照组 1.910 2.21
0.1％剂量组 2.359 2.01
0.5％剂量组 2.410 1.89
1％剂量组 2.405 1.73
5％剂量组 2.369 1.83
10％剂量组 2.391 1.77
市售丁酸梭菌产品1 1.975 2.12
市售丁酸梭菌产品2 1.902 2.25
市售丁酸梭菌产品3 1.968 2.43
试验中还观察了对罗曼褐壳蛋鸡产蛋情况影响(表21)。结果显示实验处理组明显优于添加抗生素的基础日粮对照组，也优于现有技术CN 201310083900.2公开了一株用于提高鸡蛋产率达到93.1％的丁酸梭菌，也优于现有市售丁酸梭菌产品。
表21 丁酸梭菌UCN-12发酵培养物对罗曼褐壳蛋鸡产蛋情况影响
处理组 产蛋率(％) 正常蛋率(％) 斑壳蛋率(％)
对照组 90 85 10
0.1％剂量组 97 94.7 0
0.5％剂量组 100 100 0
1％剂量组 100 100 0
5％剂量组 100 100 0
10％剂量组 100 100 0
市售丁酸梭菌产品1 91 89 8
市售丁酸梭菌产品2 93 88 10
市售丁酸梭菌产品3 90 90 10
应用实施例15 提高猪生产性能
试验选择重量相近的普通断乳仔猪70只，随机分成7组，每组10只，分别于同一养殖大棚。对照组饲喂添加抗生素的基础日粮，试验组分别在基础日粮中添加0.5％、2％和5.0％的丁酸梭菌UCN-12。自由采食和饮水，自然通风，白天自然光照，按常规接种疫苗。试验 期为180天，测定并计算增重、肉料比和死亡情况(表22)。结果显示实验处理组增重、肉料比和死亡数明显优于添加抗生素的基础日粮对照组，也优于现有市售丁酸梭菌产品。
表22 丁酸梭菌UCN-13发酵培养物对猪生长性能影响
处理组 增重(kg) 肉料比 死亡数(只)
对照组 86 2.21 3
0.5％剂量组 111 2.01 0
2％剂量组 110 1.89 0
5％剂量组 118 1.83 0
市售丁酸梭菌产品1 96 2.13 1
市售丁酸梭菌产品2 92 2.04 0
市售丁酸梭菌产品3 100 1.94 1
应用实施例16 改善鸡便臭
试验选择出生重量相近的1日龄白羽肉鸡1000只，随机分成10组，每组100只，分别于同一养殖大棚。对照组饲喂添加抗生素的基础日粮，高中低剂量试验组分别在基础日粮中添加0.1％、0.5％、1.0％、5.0％和10％的丁酸梭菌UCN-12。自由采食和饮水，自然通风，白天自然光照，晚间补充灯光照明，按常规接种疫苗，试验期为49天。采取人闻打分方法评判动物粪便臭味情况(表23)。结果显示实验处理组明显优于添加抗生素的基础日粮对照组，也优于现有市售丁酸梭菌产品。
表23 丁酸梭菌UCN-12发酵培养物对白羽肉鸡便臭的影响
处理组 便臭程度得分 备注
对照组 95 极臭
0.1％剂量组 60 略臭
0.5％剂量组 50 略臭
1％剂量组 30 略臭
5％剂量组 30 很臭
10％剂量组 30 很臭
市售丁酸梭菌产品1 85 很臭
市售丁酸梭菌产品2 85 极臭
市售丁酸梭菌产品3 90 略臭
应用实施例17 改善猪便臭
试验选择重量相近的普通断乳仔猪70只，随机分成7组，每组10只，分别于同一养殖大棚。对照组饲喂添加抗生素的基础日粮，试验组分别在基础日粮中添加0.5％、2％和5.0％的丁酸梭菌UCN-12。自由采食和饮水，自然通风，白天自然光照，按常规接种疫苗。试验期为180天，采取人闻打分方法评判动物粪便臭味情况(表24)。结果显示实验处理明显优于添加抗生素的基础日粮对照组，也优于现有市售丁酸梭菌产品。
表24 丁酸梭菌UCN-12发酵培养物对猪便臭的影响
处理组 便臭程度得分 备注
对照组 95 极臭
0.5％剂量组 40 略臭
2％剂量组 30 略臭
5％剂量组 30 略臭
市售丁酸梭菌产品1 85 很臭
市售丁酸梭菌产品2 80 很臭
市售丁酸梭菌产品3 80 很臭
应用实施例18 改善羊便臭
试验选择重量相近90日龄羊70只，随机分成7组，每组70只， 分别于同一养殖大棚。对照组饲喂添加抗生素的基础日粮，试验组分别在基础日粮中添加0.5％、2％和5.0％的丁酸梭菌UCN-12。自由采食和饮水，自然通风，白天自然光照，按常规接种疫苗。试验期为180天，采取人闻打分方法评判动物粪便臭味情况(表25)。结果显示实验处理明显优于添加抗生素的基础日粮对照组，也优于现有市售丁酸梭菌产品。
表25 丁酸梭菌UCN-12发酵培养物对羊便臭的影响
处理组 便臭程度得分 备注
对照组 100 极臭
0.5％剂量组 50 略臭
2％剂量组 30 略臭
5％剂量组 30 略臭
市售丁酸梭菌产品1 80 很臭
市售丁酸梭菌产品2 85 很臭
市售丁酸梭菌产品3 80 很臭
试验中，还发现丁酸梭菌UCN-12对不良化学物质对畜禽造成的肝脏损伤有保护作用，还能排除动物体内内毒素，也适合畜禽定期保健。
本发明有效克服了现有技术缺陷，经成都和重庆两地10家养殖企业应用，对魏氏梭菌病治疗和/或预防效果极佳，畜禽死亡率降低为0。畜禽肉品质全部为优质。禽蛋风味独特，质量优异，产品供不应求，价格几乎翻番。养殖场新增效益达到20-30％，应用示范获非常满意效果，深受养殖户欢迎。
综上所述，采用本发明菌株的发酵培养物、发酵培养物离心所得的发酵液或菌体、以及该发酵液或菌体的干燥物为有效成分喂养动 物，能治疗和/或预防魏氏梭菌病，全过程无需使用抗生素，避免了抗生素残留影响人体健康，彻底解除畜禽养殖中因大量使用抗生素对环境的污染，还可十分明显地提高畜禽的生产性能和品质，减少畜禽动物便臭，实现动物养殖环境友好，本发明工业应用前景十分看好。

Claims (9)

  1. 丁酸梭菌UCN-12菌株，其保藏编号为CGMCC No.9652。
  2. 组合物，其特征在于：以有效量的丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物为有效成分，加入饲料科学可接受的辅料制成。
  3. 根据权利要求2所述的组合物，其特征在于：所述组合物为液体或固体。
  4. 根据权利要求3所述的组合物，其特征在于：所述组合物为片剂、颗粒剂、粉剂、溶液、混悬液、乳剂或预混剂。
  5. 丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于制备治疗或预防畜禽魏氏梭菌病的兽药、饲料添加剂和/或饲料中的应用。
  6. 丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于制备降低禽蛋中胆固醇和脂肪，去除禽蛋腥味，改善禽蛋风味的兽药、饲料添加剂和/或饲料中的应用。
  7. 丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于提高畜禽的生产性能的兽药、饲料添加剂和/或饲料。
  8. 丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、 该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于制备治疗或预防母猪产前产后便秘的兽药、饲料添加剂和/或饲料中的应用。
  9. 丁酸梭菌UCN-12菌株、丁酸梭菌UCN-12菌株的发酵培养物、该发酵培养物离心所得的培养液或其干燥物，该发酵培养物离心所得的菌体或其干燥物用于制备改善畜禽便臭的兽药、饲料添加剂和/或饲料中的应用。
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Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9938558B2 (en) 2015-06-25 2018-04-10 Ascus Biosciences, Inc. Methods, apparatuses, and systems for analyzing microorganism strains from complex heterogeneous communities, predicting and identifying functional relationships and interactions thereof, and selecting and synthesizing microbial ensembles based thereon
US9993507B2 (en) 2016-01-07 2018-06-12 Ascus Biosciences, Inc. Methods of improving milk production and compositional characteristics
CN110090231A (zh) * 2018-01-30 2019-08-06 青岛东海药业有限公司 酪酸梭菌在制备预防或治疗胆管癌制剂中的应用
CN110810662A (zh) * 2019-10-09 2020-02-21 北京首朗生物科技有限公司 一种乙醇梭菌蛋白在海水肉食性鱼类中的应用
US10844419B2 (en) 2015-06-25 2020-11-24 Native Microbials, Inc. Methods, apparatuses, and systems for analyzing microorganism strains from complex heterogeneous communities, predicting and identifying functional relationships and interactions thereof, and selecting and synthesizing microbial ensembles based thereon
US10851399B2 (en) 2015-06-25 2020-12-01 Native Microbials, Inc. Methods, apparatuses, and systems for microorganism strain analysis of complex heterogeneous communities, predicting and identifying functional relationships and interactions thereof, and selecting and synthesizing microbial ensembles based thereon
CN112868928A (zh) * 2021-02-09 2021-06-01 中国农业大学 香味剂在调节母猪肠道微生物和提高繁殖性能中的应用
US11044924B2 (en) 2017-04-28 2021-06-29 Native Microbials, Inc. Methods for supporting grain intensive and or energy intensive diets in ruminants by administration of a synthetic bioensemble of microbes or purified strains therefor
CN114231434A (zh) * 2021-10-13 2022-03-25 南宁市拜欧生物工程有限责任公司 一种用于饲用添加剂的丁酸梭菌及其应用
CN114874949A (zh) * 2022-06-06 2022-08-09 江苏三仪生物工程有限公司 一种丁酸梭菌、其发酵物、包含其的菌剂及动物饲料添加剂
CN115141781A (zh) * 2022-05-07 2022-10-04 青岛农业大学 一种提高凝结芽孢杆菌中亚铁离子含量的方法和复合菌剂
US11891647B2 (en) 2016-12-28 2024-02-06 Native Microbials, Inc. Methods, apparatuses, and systems for analyzing complete microorganism strains in complex heterogeneous communities, determining functional relationships and interactions thereof, and identifying and synthesizing bioreactive modificators based thereon
CN117617353A (zh) * 2024-01-09 2024-03-01 广东省农业科学院动物科学研究所 一种农业废弃物资源化利用方法、添加剂及其制备方法

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104611273B (zh) * 2015-02-11 2017-10-31 成都与康科技有限公司 丁酸梭菌ucn‑12菌株及其组合物和应用
CN107254424B (zh) * 2017-07-11 2023-02-28 福建贝迪药业有限公司 一种畜禽用新型液态复合微生态制剂及其制备方法
CN107312730B (zh) * 2017-07-11 2023-02-28 福建贝迪药业有限公司 一种复方中草药微生态制剂及其基于固态发酵的快速生产方法
CN110150486A (zh) * 2018-04-03 2019-08-23 湖南景程电子科技有限公司 一种防治哺乳母猪便秘腹泻等肠道疾病的饲料添加剂
CN110951640A (zh) * 2019-12-06 2020-04-03 山东蔚蓝生物科技有限公司 一种益生丁酸梭菌及其应用
CN111493237A (zh) * 2020-05-09 2020-08-07 湖北绿科乐华生物科技有限公司 一种改善鸡蛋品质的蛋鸡饲料添加剂组合
CN112655827A (zh) * 2020-12-21 2021-04-16 广东溢多利生物科技股份有限公司 反刍动物专用丁酸梭菌制剂及其制备方法和应用
CN112458030A (zh) * 2020-12-22 2021-03-09 广东溢多利生物科技股份有限公司 一种反刍动物专用丁酸梭菌制剂的制备方法

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101942405A (zh) * 2010-07-27 2011-01-12 湖北工业大学 一种己酸菌
CN102670663A (zh) * 2008-01-31 2012-09-19 青岛东海药业有限公司 梭菌制剂及其应用
CN102919570A (zh) * 2012-10-25 2013-02-13 淮安正昌饲料有限公司 一种怀孕后期母猪专用攻胎配合饲料及其制备方法
CN104277999A (zh) * 2014-09-15 2015-01-14 中国农业大学 一种新型多效丁酸梭菌及其在提高动物抗氧化能力和改善肉品质方面的应用
CN104611273A (zh) * 2015-02-11 2015-05-13 成都与康科技有限公司 丁酸梭菌ucn-12菌株及其组合物和应用

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101095698B (zh) * 2006-06-26 2010-12-01 青岛东海药业有限公司 酪酸梭菌防治便臭毒素引起的相关症状和疾病的用途
CN102319270B (zh) * 2011-09-26 2012-10-17 湖北绿雪生物产业有限公司 采用补料分批发酵法制备高密度酪酸菌活菌制剂的方法
KR101633610B1 (ko) * 2011-12-14 2016-06-28 일동제약주식회사 자가분해효소를 제거한 유포자 프로바이오틱스의 마이크로캡슐화 방법 및 그 방법으로 제조된 유포자 프로바이오틱스

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102670663A (zh) * 2008-01-31 2012-09-19 青岛东海药业有限公司 梭菌制剂及其应用
CN101942405A (zh) * 2010-07-27 2011-01-12 湖北工业大学 一种己酸菌
CN102919570A (zh) * 2012-10-25 2013-02-13 淮安正昌饲料有限公司 一种怀孕后期母猪专用攻胎配合饲料及其制备方法
CN104277999A (zh) * 2014-09-15 2015-01-14 中国农业大学 一种新型多效丁酸梭菌及其在提高动物抗氧化能力和改善肉品质方面的应用
CN104611273A (zh) * 2015-02-11 2015-05-13 成都与康科技有限公司 丁酸梭菌ucn-12菌株及其组合物和应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
YI, ZHONGHUA;: "Research and Application Progress of Feeding Microbioecologics Clostridium Butyricum", FEED RESEARCH, vol. 2, 29 February 2012 (2012-02-29), pages 14 - 17, ISSN: 1002-2813 *

Cited By (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10844419B2 (en) 2015-06-25 2020-11-24 Native Microbials, Inc. Methods, apparatuses, and systems for analyzing microorganism strains from complex heterogeneous communities, predicting and identifying functional relationships and interactions thereof, and selecting and synthesizing microbial ensembles based thereon
US9938558B2 (en) 2015-06-25 2018-04-10 Ascus Biosciences, Inc. Methods, apparatuses, and systems for analyzing microorganism strains from complex heterogeneous communities, predicting and identifying functional relationships and interactions thereof, and selecting and synthesizing microbial ensembles based thereon
US10851399B2 (en) 2015-06-25 2020-12-01 Native Microbials, Inc. Methods, apparatuses, and systems for microorganism strain analysis of complex heterogeneous communities, predicting and identifying functional relationships and interactions thereof, and selecting and synthesizing microbial ensembles based thereon
US10966437B2 (en) 2016-01-07 2021-04-06 Native Microbials, Inc. Microbial compositions and methods of use for improving milk production
US11291219B2 (en) 2016-01-07 2022-04-05 Native Microbials, Inc. Microbial compositions and methods of use for improving milk production
US10448658B2 (en) 2016-01-07 2019-10-22 Ascus Biosciences, Inc. Cow food and methods of husbandry for increased milk production
US10448657B2 (en) 2016-01-07 2019-10-22 Ascus Biosciences, Inc. Cow food and methods of husbandry for increased milk production
US11910808B2 (en) 2016-01-07 2024-02-27 Native Microbials, Inc. Ruminant compositions
US10645952B2 (en) 2016-01-07 2020-05-12 Ascus Biosciences, Inc. Microbial compositions and methods of use for improving milk production
US10701955B2 (en) 2016-01-07 2020-07-07 Ascus Biosciences, Inc. Ruminant compositions
US11910809B2 (en) 2016-01-07 2024-02-27 Native Microbials, Inc. Microbial compositions and methods of use for improving milk production
US10293006B2 (en) 2016-01-07 2019-05-21 Ascus Biosciences, Inc. Microbial compositions for improving milk production in ruminants
US9993507B2 (en) 2016-01-07 2018-06-12 Ascus Biosciences, Inc. Methods of improving milk production and compositional characteristics
US10398154B2 (en) 2016-01-07 2019-09-03 Ascus Biosciences, Inc. Microbial compositions and methods of use for improving milk production
US11891647B2 (en) 2016-12-28 2024-02-06 Native Microbials, Inc. Methods, apparatuses, and systems for analyzing complete microorganism strains in complex heterogeneous communities, determining functional relationships and interactions thereof, and identifying and synthesizing bioreactive modificators based thereon
US11044924B2 (en) 2017-04-28 2021-06-29 Native Microbials, Inc. Methods for supporting grain intensive and or energy intensive diets in ruminants by administration of a synthetic bioensemble of microbes or purified strains therefor
US11871767B2 (en) 2017-04-28 2024-01-16 Native Microbials, Inc. Microbial compositions and methods for ruminant health and performance
CN110090231A (zh) * 2018-01-30 2019-08-06 青岛东海药业有限公司 酪酸梭菌在制备预防或治疗胆管癌制剂中的应用
CN110810662A (zh) * 2019-10-09 2020-02-21 北京首朗生物科技有限公司 一种乙醇梭菌蛋白在海水肉食性鱼类中的应用
CN112868928A (zh) * 2021-02-09 2021-06-01 中国农业大学 香味剂在调节母猪肠道微生物和提高繁殖性能中的应用
CN114231434A (zh) * 2021-10-13 2022-03-25 南宁市拜欧生物工程有限责任公司 一种用于饲用添加剂的丁酸梭菌及其应用
CN115141781A (zh) * 2022-05-07 2022-10-04 青岛农业大学 一种提高凝结芽孢杆菌中亚铁离子含量的方法和复合菌剂
CN115141781B (zh) * 2022-05-07 2023-06-16 青岛农业大学 一种提高凝结芽孢杆菌中亚铁离子含量的方法和复合菌剂
CN114874949A (zh) * 2022-06-06 2022-08-09 江苏三仪生物工程有限公司 一种丁酸梭菌、其发酵物、包含其的菌剂及动物饲料添加剂
CN114874949B (zh) * 2022-06-06 2023-09-12 江苏三仪生物工程有限公司 一种丁酸梭菌、其发酵物、包含其的菌剂及动物饲料添加剂
CN117617353A (zh) * 2024-01-09 2024-03-01 广东省农业科学院动物科学研究所 一种农业废弃物资源化利用方法、添加剂及其制备方法

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