CN113967243B - Fermented traditional Chinese medicine preparation for preventing and treating diarrhea of poultry and preparation method thereof - Google Patents
Fermented traditional Chinese medicine preparation for preventing and treating diarrhea of poultry and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/75—Rutaceae (Rue family)
- A61K36/756—Phellodendron, e.g. corktree
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/284—Atractylodes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/481—Astragalus (milkvetch)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/57—Magnoliaceae (Magnolia family)
- A61K36/575—Magnolia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/734—Crataegus (hawthorn)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/736—Prunus, e.g. plum, cherry, peach, apricot or almond
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/76—Salicaceae (Willow family), e.g. poplar
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/906—Zingiberaceae (Ginger family)
- A61K36/9068—Zingiber, e.g. garden ginger
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/12—Antidiarrhoeals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Abstract
The invention discloses a fermented traditional Chinese medicine preparation for preventing and treating diarrhea of poultry and a preparation method thereof. The traditional Chinese medicine preparation is prepared by fermenting the following raw materials: bran, bean pulp, traditional Chinese medicine mixed powder, zymophyte liquid and water. The invention can be widely applied to preventing and treating diarrhea of poultry, can effectively improve the immunity of the poultry, and can also avoid the problems of drug resistance and veterinary drug residues caused by pathogenic bacteria. The traditional Chinese medicine preparation has reasonable formula, easily obtained medicinal materials, simple preparation process and low production cost, and is beneficial to expanded production.
Description
Technical Field
The invention belongs to the technical field of veterinary drug fermentation, and in particular relates to a fermented traditional Chinese medicine preparation for preventing and treating diarrhea of poultry and a preparation method thereof.
Background
The Chinese herbal medicine is a valuable cultural heritage in China, is an intelligent crystal for struggling with diseases of Chinese nationality for thousands of years, and is increasingly valued as a new Chinese medicine processing method along with the deep research of Chinese herbal medicine preparations, and compared with the Chinese medicine, the fermented Chinese medicine has obvious advantages. The modern traditional Chinese medicine fermentation technology, also called traditional Chinese medicine bioconversion technology, is a new technology of traditional Chinese medicine modernization pharmacy formed by combining the subjects of modern biotechnology and the like on the basis of inheriting the traditional Chinese medicine fermentation processing method. The research of modern traditional Chinese medicine fermentation technology starts in the 80 s of the 20 th century, and the research of traditional Chinese medicine fermentation technology is mostly single traditional Chinese medicine fermentation, and mainly focuses on fungus self fermentation, such as ganoderma lucidum and cordyceps sinensis mycelium fermentation to produce secondary metabolites. At present, the research of traditional Chinese medicine fermentation has been developed from single medicine to compound medicine, and the fermented traditional Chinese medicine preparation can change the medicine property to a greater extent, improve the curative effect, reduce the toxic and side effects and enlarge the indication. They can decompose and convert the drugs into new active ingredients, and the new drug effect is mainly produced in the following 5 aspects: (1) taking the active ingredients as precursors, and forming new compounds through microbial metabolism; (2) the secondary metabolite produced by the growth of the microorganism contains the medicine with good efficacy; (3) the secondary metabolite can produce synergistic effect with the effective components of the traditional Chinese medicine; (4) certain substances of the traditional Chinese medicine can promote or inhibit the growth and metabolism of microorganisms and active ingredients, and the microorganisms can change own metabolic pathways to form new active ingredients or change the mutual proportion of the active ingredients; (5) the microorganism can decompose toxic substances of the traditional Chinese medicine and reduce toxic and side effects of the medicine; (6) the microorganism can convert macromolecular substances into small molecules which are easy to absorb, so that the effective components in blood can reach the effective concentration rapidly. Therefore, the processed traditional Chinese medicine has advantages over the common physical or chemical means. From the development history of traditional Chinese medicine fermentation, the fermentation is the same as the traditional food fermentation, and some varieties even serve as food and medicine. Since research on dominant strain separation identification and biochemical reaction mechanism in fermentation process is the basis for explaining the traditional Chinese medicine fermentation principle, if the technology and the achievement of strain identification and fermentation mechanism research of food fermentation are introduced into traditional Chinese medicine fermentation, the research level of the technology and the achievement of fermentation mechanism research is improved.
With the rapid development of the cultivation in China, the spread and outbreak of seasonal, epidemic and infectious diseases of poultry and livestock are more frequently seen, especially in the north of early spring, autumn and winter, the temperature difference between day and night is large, cold and dry, and various complex environmental factors are added, so that various diseases are often caused, the cultivation industry is greatly damaged, whether the digestive tract diseases or the respiratory tract diseases are usually represented as weak resistance, slow growth, increased morbidity and mortality of animals, one of the main factors restricting the development of the animal industry is also one of the reasons causing economic losses of farmers.
Lactobacillus plantarum belongs to gram-positive bacteria and is an anaerobic or facultative anaerobic bacterium. Is widely existed in the nature and also exists in the human gastrointestinal tract, plays a beneficial role through the stomach and is planted in the intestinal tract, and is a probiotic flora in the human gastrointestinal tract. At present, a great deal of researches show that lactobacillus plantarum has high safety and various probiotic functions such as regulating intestinal flora balance, improving gastrointestinal tract functions, participating in immune system regulation and the like.
The bacillus is an internationally accepted microorganism strain which can be directly added into word materials, has the characteristics of no toxicity and no residue, and the bacillus subtilis has the characteristics of acid resistance, high temperature resistance and the like, and can be widely applied to industrial word materials. The probiotics mainly comprising bacillus subtilis and the prebiotics represented by oligosaccharide have the functions of regulating the microecological balance of animal, improving the digestion utilization rate of word materials, promoting the improvement of the immune function of organisms and the like. The bacillus with low-temperature culture and strong stress resistance is used for influencing the production performance and immune function of broiler chickens, so that the effect of the broiler chickens is stable, and the effect of the broiler chickens can be ensured under certain adverse conditions such as adverse conditions and indistinct medication conditions. The microecological environment of animal organism is affected by many factors, and the intestinal microecological flora is different under the conditions of different varieties, different feeding environments, different nutrition levels, different feed compositions and the like. The preparation containing the bacillus subtilis has high pertinence to the breeding industry which needs green and environment protection in China.
Under the current high-density cultivation and repeated drug application cultivation environment, the resistance of the poultry body is relatively low due to the influence of a plurality of factors such as environment, drug application and the like, the occurrence of intestinal diseases can cause the reduction of egg yield and meat yield, the method is a very serious test for the cultivation industry, and as the harm of abusing antibiotics is more serious, no resistance and no replacement resistance become the current research hot spot. Research on traditional Chinese medicines by probiotic fermentation is getting more and more important, and the traditional Chinese medicines after fermentation treatment are rich in vitamins, enzymes, growth factors and the like. Has the characteristics of green and high efficiency, and has the advantages of difficult generation of drug resistance and difficult generation of dysbacteriosis. Therefore, development of a fermented traditional Chinese medicine preparation for preventing and treating diarrhea is urgent.
Disclosure of Invention
Based on the reasons, the invention combines the traditional theory of traditional Chinese medicine and Chinese veterinarian and the related research of microecologics to develop a new Chinese veterinary medicine preparation, provides a fermented traditional Chinese medicine preparation for preventing and treating diarrhea of poultry, and has the advantages of convenient administration, safety, reliability, immunity enhancement, digestion promotion, and intestinal tract harmful bacteria inhibition.
The invention is realized mainly by the following technical scheme.
In one aspect of the invention, a fermented traditional Chinese medicine preparation for preventing and treating diarrhea of poultry is provided, and the preparation is formed by fermenting a fermentation substrate and fermentation bacteria liquid.
The addition amount of the fermentation bacteria liquid is 2% -5% of the total amount of the fermentation substrate. Preferably, the addition amount of the fermentation broth is 2.5% of the total amount of the fermentation substrate.
The fermentation substrate consists of bran, bean pulp and traditional Chinese medicine mixed powder, wherein the weight percentages of the components are as follows: 10-30% of bran, 10-30% of bean pulp and 40-70% of traditional Chinese medicine mixed powder.
Preferably, the weight percentage of each component in the fermentation substrate is as follows: 20-30% of bran, 20-30% of bean pulp and 40-60% of traditional Chinese medicine mixed powder.
More preferably, the fermentation substrate comprises the following components in percentage by weight: 20% of bran, 20% of bean pulp and 60% of traditional Chinese medicine mixed powder.
The traditional Chinese medicine mixed powder comprises the following traditional Chinese medicine components in parts by weight: 20-50 parts of amur corktree bark, 10-30 parts of dark plum fruit, 50-100 parts of poplar flower, 20-50 parts of purslane, 10-30 parts of dried ginger, 30-50 parts of largehead atractylodes rhizome, 30-50 parts of officinal magnolia bark, 50-100 parts of medicated leaven, 10-30 parts of hawthorn fruit, 10-30 parts of myrobalan and 30-50 parts of astragalus membranaceus.
Preferably, the traditional Chinese medicine mixed powder comprises the following traditional Chinese medicine components in parts by weight: 30 parts of phellodendron, 20 parts of dark plum, 70 parts of poplar flower, 30 parts of purslane, 20 parts of dried ginger, 40 parts of bighead atractylodes rhizome, 40 parts of magnolia officinalis, 70 parts of medicated leaven, 20 parts of hawthorn, 20 parts of myrobalan and 40 parts of astragalus membranaceus.
The fermentation broth contains lactobacillus plantarum 1×10 8～9 cfu/mL, bacillus subtilis 1×10 8～9 cfu/mL, microzyme 1×10 8～9 cfu/mL。
Preferably, the lactobacillus plantarum is lactobacillus plantarum RP1, is classified and named as lactobacillus plantarum (Lactobacillus plantarum), has a strain number of RP1 and is preserved in China center for type culture Collection, with a preservation address of university of Wuhan and a preservation number of CCTCC NO: m2018802, date of preservation 2018.11.19.
In another aspect of the invention, a preparation method of a fermented traditional Chinese medicine preparation is provided, which comprises the following specific steps:
(1) Preparing fermentation bacteria liquid: inoculating the strains into special culture mediums respectively, culturing seed solutions according to the special conditions, and stopping culturing when the strains grow to logarithmic phase and are transferred into a fermentation tank for expansion culture respectively and reach the logarithmic phase.
(2) The preparation of the traditional Chinese medicine mixed powder comprises the following steps: removing impurities from the medicinal materials respectively, crushing the medicinal materials to 80-100 meshes, and mixing the medicinal materials according to the weight parts.
(3) Removing impurities from bran and bean pulp, pulverizing in a pulverizer, taking out, placing in a mixer, and mixing the prepared Chinese medicinal powder.
(4) Spraying fermentation bacteria liquid into a mixer for mixing, adding a certain amount of water, fully and uniformly mixing, transferring the obtained material to a discharging machine, accurately weighing, filling into a fermentation bag with a one-way breather valve, sealing, and placing into a fermentation chamber for fermentation.
The fermentation conditions in the fermentation chamber are as follows: and fermenting at room temperature of 25-37 deg.c and humidity of 50-70% for 7-14 days.
Preferably, the fermentation conditions in the fermentation chamber are: the room temperature is 28 ℃, the humidity is 65 percent, and the fermentation is carried out for 12 days.
The amount of the added water in the step (4) is controlled to be 40-60% of the total water content.
Preferably, the amount of added water in step (4) is controlled to 55% of the total water content.
The fermented traditional Chinese medicine preparation for preventing and treating diarrhea of poultry can be used for preparing feed additives.
The beneficial effects are that:
1. the mixed strain in the fermentation broth can be quickly propagated into dominant strain, so that the raw materials can be inoculated for fermentation without sterilization.
2. The probiotics in the invention can generate a large amount of enzymes in the fermentation process, and can promote the dissolution rate of active ingredients in the traditional Chinese medicinal materials, thereby improving the medicinal effect.
3. The traditional Chinese medicine composition contains components capable of obviously inhibiting bacteria, plays a role in inhibiting harmful intestinal flora, and has the prospect of reducing and replacing resistance.
4. The fermented traditional Chinese medicine preparation can obviously improve the diarrhea condition of broiler chickens caused by escherichia coli and reduce the death rate.
5. The fermented traditional Chinese medicine preparation can improve immunity, improve production performance and reduce diarrhea rate, so that disease resistance of animal organisms is improved.
Detailed Description
The foregoing will be described in further detail by way of the following detailed description of the embodiments. It should not be construed that the scope of the above subject matter is limited to the following examples. All techniques implemented based on this disclosure are within the scope.
Example 1
A fermented Chinese medicinal preparation for preventing and treating diarrhea of poultry is prepared by fermenting fermentation substrate and fermentation bacteria liquid, wherein the fermentation substrate contains Chinese medicinal mixed powder 60%, bran 20% and soybean meal 20%. The traditional Chinese medicine mixed powder comprises the following traditional Chinese medicine components: 30 parts of phellodendron, 20 parts of dark plum, 70 parts of poplar flower, 30 parts of purslane, 20 parts of dried ginger, 40 parts of bighead atractylodes rhizome, 40 parts of magnolia officinalis, 70 parts of medicated leaven, 20 parts of hawthorn, 20 parts of myrobalan and 40 parts of astragalus membranaceus. The specific preparation method of the fermented traditional Chinese medicine preparation comprises the following steps:
(1) Preparing fermentation bacteria liquid: inoculating lactobacillus plantarum, bacillus subtilis and saccharomycetes into special culture mediums respectively, culturing seed solutions according to the special conditions, and stopping culturing when the strains grow to the logarithmic growth phase and are respectively transferred into a fermentation tank for expansion culture and reach the logarithmic growth phase.
(2) The preparation of the traditional Chinese medicine mixed powder comprises the following steps: removing impurities from the medicinal materials respectively, crushing the medicinal materials to 80-100 meshes, and uniformly mixing the medicinal materials according to the weight parts.
(3) Removing impurities from bran and bean pulp, pulverizing in a pulverizer, taking out, placing in a mixer, and mixing the prepared Chinese medicinal powder.
(4) Fermenting bacteria liquid accounting for 2.5% of the total amount of fermentation substrate (the concentration of each bacteria is controlled to be 1×10) 9 cfu/mL) is sprayed into a mixer for mixing, a certain amount of water (the water adding amount is controlled to be 55% of the whole water content) is added, the obtained material is transferred to a discharging machine after fully and uniformly mixing, and after accurately weighing, the material is filled into a fermentation bag with a one-way breather valve and sealed, and is placed into a fermentation chamber for fermentation for 12 days at the room temperature of 28 ℃ and the humidity of 65%.
Example 2
A fermented Chinese medicinal preparation for preventing and treating diarrhea of poultry is prepared by fermenting fermentation substrate and fermentation bacteria liquid, wherein the fermentation substrate contains 40% of Chinese medicinal mixed powder, 30% of bran and 30% of soybean meal. The traditional Chinese medicine mixed powder comprises the following traditional Chinese medicine components: 20 parts of phellodendron, 10 parts of dark plum, 50 parts of poplar flower, 20 parts of purslane, 10 parts of dried ginger, 30 parts of bighead atractylodes rhizome, 30 parts of magnolia officinalis, 50 parts of medicated leaven, 10 parts of hawthorn, 10 parts of myrobalan and 30 parts of astragalus membranaceus. The specific preparation method of the fermented traditional Chinese medicine preparation comprises the following steps:
(1) Preparing fermentation bacteria liquid: inoculating lactobacillus plantarum, bacillus subtilis and saccharomycetes into special culture mediums respectively, culturing seed solutions according to the special conditions, and stopping culturing when the strains grow to the logarithmic growth phase and are respectively transferred into a fermentation tank for expansion culture and reach the logarithmic growth phase.
(2) The preparation of the traditional Chinese medicine mixed powder comprises the following steps: removing impurities from the medicinal materials respectively, crushing the medicinal materials to 80-100 meshes, and uniformly mixing the medicinal materials according to the weight parts.
(3) Removing impurities from bran and bean pulp, pulverizing in a pulverizer, taking out, placing in a mixer, and mixing the prepared Chinese medicinal powder.
(4) Fermenting bacteria solution (concentration of each bacteria is controlled to be 1×10) accounting for 3.5% of total fermentation substrate 9 cfu/mL) is sprayed into a mixer for mixing, a certain amount of water (the water adding amount is controlled to be 45% of the whole water content) is added, the obtained material is transferred to a discharging machine after fully and uniformly mixing, and after accurately weighing, the material is filled into a fermentation bag with a one-way breather valve and sealed, and is placed into a fermentation chamber for fermentation for 7 days at the room temperature of 25 ℃ and the humidity of 50%.
Example 3
A fermented Chinese medicinal preparation for preventing and treating diarrhea of poultry is prepared by fermenting fermentation substrate and fermentation bacteria liquid, wherein the fermentation substrate contains Chinese medicinal mixed powder 70%, bran 10% and soybean meal 20%. The traditional Chinese medicine mixed powder comprises the following traditional Chinese medicine components: 50 parts of phellodendron, 30 parts of dark plum, 100 parts of poplar flower, 50 parts of purslane, 30 parts of dried ginger, 50 parts of bighead atractylodes rhizome, 50 parts of magnolia officinalis, 100 parts of medicated leaven, 30 parts of hawthorn, 30 parts of myrobalan and 50 parts of astragalus membranaceus. The specific preparation method of the fermented traditional Chinese medicine preparation comprises the following steps:
(1) Preparing fermentation bacteria liquid: inoculating lactobacillus plantarum, bacillus subtilis and saccharomycetes into special culture mediums respectively, culturing seed solutions according to the special conditions, and stopping culturing when the strains grow to the logarithmic growth phase and are respectively transferred into a fermentation tank for expansion culture and reach the logarithmic growth phase.
(2) The preparation of the traditional Chinese medicine mixed powder comprises the following steps: removing impurities from the medicinal materials respectively, crushing the medicinal materials to 80-100 meshes, and uniformly mixing the medicinal materials according to the weight parts.
(3) Removing impurities from bran and bean pulp, pulverizing in a pulverizer, taking out, placing in a mixer, and mixing the prepared Chinese medicinal powder.
(4) Fermenting bacteria solution accounting for 4.5% of the total amount of fermentation substrate (the concentration of each bacteria is controlled to be 1×10) 9 cfu/mL) is sprayed into a mixer for mixing, a certain amount of water (the water adding amount is controlled to be 60% of the whole water content) is added, the obtained material is transferred to a discharging machine after fully and uniformly mixing, and after accurately weighing, the material is filled into a fermentation bag with a one-way breather valve and sealed, and is placed into a fermentation chamber for fermentation for 14 days at the room temperature of 37 ℃ and the humidity of 70%.
Example 4 extract assay
Extract assay according to the invention
1. Test purpose: it was determined whether the extract content of the present invention increased after fermentation.
2. Test protocol
2.1 test materials
Purified water, ethanol, experimental group 1 (sample of example 1 without fermentation), experimental group 2 (sample of example 1 according to the invention), experimental group 3 (comparative group of different species in fermentation broth).
The comparative example group is: the strain in the fermentation broth is a sample of example 1 consisting of lactobacillus plantarum, bacillus subtilis, saccharomycetes and enterococcus faecalis, and other conditions are unchanged.
2.2 test procedure
(1) Water soluble extract content: the test group is placed in a 60 ℃ oven for drying, about 2-4 g of the test sample is taken, the test sample is precisely formed, the test sample is placed in a 250ml conical flask, 100ml of water is precisely added, the test sample is sealed, the test sample is weighed, the test sample is placed for 1 hour, a reflux condenser is connected, the test sample is heated to boiling, and the test sample is kept slightly boiling for 1 hour. Taking down the conical flask after cooling, sealing, weighing again, supplementing the lost weight with water, shaking uniformly, filtering with a drying filter, precisely measuring 25ml of filtrate, placing the filtrate in a dried evaporation dish with constant weight, evaporating in water bath, drying for 3 hours at 105 ℃, placing the dried product in a dryer for cooling again for 30 minutes, and rapidly and precisely weighing. The water-soluble extract content (%) in the formulation was calculated as a dry product unless otherwise specified.
(2) Alcohol extract content: the experimental group is placed in a 60 ℃ oven for drying, about 2-4 g of the sample to be tested is taken, the sample is precisely formed, the sample is placed in a 250ml conical flask, 100ml of ethanol is precisely added, the sample is sealed, the sample is weighed, the sample is kept stand for 1 hour, a reflux condenser tube is connected, the sample is heated to boiling in a water bath, and the sample is kept slightly boiling for 1 hour. Taking down the conical flask after cooling, sealing, weighing again, supplementing the lost weight with ethanol, shaking uniformly, filtering with a drying filter, precisely measuring 25ml of filtrate, placing the filtrate in a dried evaporation dish with constant weight, evaporating in water bath, drying for 3 hours at 105 ℃, placing the dried product in a dryer for cooling again for 30 minutes, and rapidly and precisely weighing. The alcohol-soluble extract content (%) in the formulation was calculated as a dry product unless otherwise specified.
3. Test results
| Group of | Water-soluble extract | Alcohol soluble extract |
| Experiment group 1 | 28.4％ | 12.1％ |
| Experiment group 2 | 36.1％ | 14.2％ |
| Experiment group 3 | 30.2％ | 14.3％ |
From the above experiments, it was concluded that the preferred combination of species produced a significantly higher extract content after fermentation of the pharmaceutical composition than the unfermented group and other combinations of species.
The commercial pulsatilla root oral liquid described in the following examples is pulsatilla root oral liquid used in a normal administration procedure of a certain farm, and the pulsatilla root oral liquid can be effectively used for treating diarrhea diseases.
Example 5 bacteriostasis test
The antibacterial effect test of the invention
1. Purpose of test
Verification of the bacteriostatic effect of the invention
2. Test protocol
2.1 test materials
(1) Preparation of test samples: taking 50g of each sample of the embodiment 1 and the embodiment 2, respectively adding 500ml of ultrapure water, heating and refluxing for 1h, filtering, centrifuging at 3000rpm for 10min, taking supernatant, concentrating under reduced pressure until 1ml of the supernatant contains 1g of crude drug, keeping at 4 ℃ for later use, sterilizing at high pressure before use, and cooling to normal temperature for use.
(2) Test materials: MH agar medium, clinical chicken colibacillus strain, puncher and vernier caliper.
2.2 test procedure
The experiments were divided into a negative control group, a model group, and example 1 and example 2, and a commercial white head Weng Koufu liquid group.
The sensitivity of the invention to clinically isolated E.coli was examined by a well-punch method. The coliform bacteria liquid is adjusted to be 1.5X10 8 CFU/ml, 100. Mu.L of the mixture was uniformly spread on a plated MH plate medium, and the mixture was allowed to stand aseptically for about 15 minutes. The agar in the wells is removed by uniformly punching (6 mm) 4 plates with a sterile punch, and 100. Mu.L of the Chinese medicinal liquid is added to the wells. After 16h incubation at 37 ℃, the bacteriostasis was checked, the diameter of the zone of inhibition was measured and the average value was calculated. Judging the sensitivity according to literature records: the diameter of the inhibition zone is less than 10mm, the inhibition zone is weakly sensitive, the inhibition zone is moderately sensitive, and the inhibition zone is highly sensitive when the diameter is more than 15 mm.
3. Test results
| Grouping | Diameter of inhibition zone (mm) |
| Example 1 | 13.5±0.12 |
| Example 2 | 18.2±0.24 |
| Commercial white head Weng Koufu liquid group | 12.23±0.04 |
Example 6 immune index, intestinal flora assay
Test of the Effect of the present invention on poultry immunization
1. Purpose of test
By using the invention on animal bodies, the effect influence of the invention on immunity is examined
2. Test protocol
2.1 test subjects
The test animals were 400 healthy 1 day-old broilers. The medicine is randomly divided into four groups, wherein one group is the low-dose group of the invention, two groups are the medium-dose group of the invention, three groups are the high-dose group of the invention, and four groups are the control group.
2.2 test materials
The samples of the test are provided by veterinary drug research and development centers of pharmaceutical research and development institute of the biological pharmaceutical industry, inc., and the experimental groups are low, medium and high, 500 g/bag and lot number 20191001 of the fermented traditional Chinese medicine preparation in the embodiment 2 of the invention.
2.3 test methods
400 broilers are randomly divided into 4 groups, 100 broilers are in each group, the test period is 42 days, one group is a control group, 3 groups are experimental groups, wherein a low-dose group is fed according to an additive amount of 1%o, a medium-dose group is fed according to an additive amount of 3%o, a high-dose group is fed according to an additive amount of 5%o, the control group is fed according to a conventional mode, free feeding is carried out, the feeding amount and death amount of each group are recorded every day, and weighing is carried out on 7 days, 14 days, 21 days, 28 days, 35 days and 42 days.
(1) Immune index assay: in the 42-day-old 4-group experiments, 20 healthy chickens with body weights close to the average body weight were selected for slaughter, the bursa, thymus and spleen were removed completely, weighed and recorded, and the bursa index, thymus index and spleen index of each group were calculated.
(2) Effect test of intestinal flora: in 4 groups of experiments at 42 days of age, 20 healthy chickens with weight close to average weight are selected in each group, after slaughter is carried out on an empty stomach, the left cecum is immediately taken out, 0.5g of content and asepsis audience are taken out in an ultra clean bench, 4.5ml of asepsis physiological saline is added, the mixture is vibrated for 5min by an oscillator and centrifuged for 10min (1000 r/min), the supernatant is taken and subjected to 10-time gradient dilution, and the number of the escherichia coli, the lactobacillus and the lactobacillus is measured by a plate counting method. Bacterial numbers were counted using plate colony counting and the results were expressed as logarithm of the number of bacteria per gram of advocated content (lgCFU/g).
3. Test results
(1) Immune index measurement test
| Group of | Bursal index | Thymus index | Spleen index |
| Low dose group | 1.32±0.19 a | 2.45±0.35 a | 1.35±0.54 a |
| Medium dose group | 1.48±0.18 b | 3.23±0.52 b | 1.63±0.42 b |
| High dose group | 1.39±0.22 a | 2.73±0.22 a | 1.44±0.22 a |
| Control group | 1.23±0.23 a | 2.32±0.19 a | 1.31±0.42 a |
(2) Intestinal flora effect test (lgCFU/g)
| Group of | Coli (lgCFU/g) | Lactobacillus (lgCFU/g) |
| Low dose group | 6.98±0.04a | 8.12±0.06 a |
| Medium dose group | 7.32±0.07a | 8.05±0.05 a |
| High dose group | 7.41±0.05 a | 8.13±0.08 a |
| Control group | 7.75±0.06 a | 7.56±0.04 a |
Example 7 diarrhea model efficacy validation test
The invention tests the treatment effect of the broiler chicken diarrhea caused by colibacillus
1. Purpose of test
It is verified whether the present invention can produce a therapeutic effect on a broiler model of E.coli diarrhea.
2. Test protocol
2.1 test subjects
The test selects 1 day-old immunity-free and drug-free broiler chickens, and feeds the broiler chickens to 7 days-old chickens, and the whole course uses the antibiotic-free feed. The experimental animals are randomly divided into groups, wherein one group is the group of the invention, two groups are the group of the unfermented Chinese medicinal mixed powder, three groups are positive control groups, four groups are model groups, and five groups are blank groups.
2.2 test materials
The samples of the test are provided by veterinary drug research and development centers of pharmaceutical research and development institute of the biological pharmaceutical industry, inc., of Repujin, and the experiment 1 group is 1000 g/bag of the fermentation preparation in the embodiment 1 of the invention, and lot number 20191201. The experiment 2 group is the traditional Chinese medicine mixed powder in the invention, and the positive medicine group (experiment 3 group) isCommercially available pulsatilla chinensis bunge oral liquid; the strain of Escherichia coli HuB200628 provided by Reep biopharmaceutical institute is used for tapping, and the concentration of bacterial solution is 1×10 11 CFU/ml。
2.3 test methods
2.3.1 method of Forming mold
Purchasing 1 day old broiler, feeding to 7 days old, randomly grouping (15 per group) and fin numbering, collecting HuB200628 Escherichia coli strain, and controlling bacterial liquid concentration at 1×10 8 CFU/ml, adopting the mode of gastric lavage and rectal tapping, wherein the gastric lavage dosage is 1ml each, the rectal tapping dosage is 0.5ml, tapping is carried out again at intervals of 5 hours after the first tapping, the steps are the same as above (a blank group uses sterile physiological saline to replace tapping), after diarrhea symptoms appear, weighing is carried out, diarrhea scoring is carried out, and then the medicine is administered according to the following administration mode.
2.3.2 methods of administration
Experiment 1 group: the fermented traditional Chinese medicine preparation in the embodiment 1 of the invention is used for 7 days once a day according to the proportion of 1 jin of the mixed material per 1g (one bag) of the product;
experiment 2 group: when the traditional Chinese medicine mixed powder is used, 1 jin of materials are mixed with 1g of the traditional Chinese medicine mixed powder every day for 7 days;
experiment 3 group: the commercial pulsatilla chinensis bunge oral liquid is used for 7 days.
One day after the end of the dose was observed, followed by weighing and diarrhea scoring.
2.3.3 observations index
(1) Daily observation: mainly observe animal appearance, activity condition, respiration state, fecal morphology, nutritional status, fur luster, mortality, etc.
(2) Diarrhea score: diarrhea scoring of animals daily before and after molding
The following is shown:
3. test results
3.1 overall case
The chickens begin to show symptoms 4-5 hours after the stomach irrigation and the rectum sterilization in the morning, and the symptoms are mainly manifested by mental depression, appetite reduction, rough feathers, eye closing and sleepiness, unwilling activities, incapacity of lying on the ground, white diarrhea or watery feces and the like of serious people. The dissected dead chicken can be seen to have a large amount of cheese-like substances in the abdominal cavity, and the phenomena of pericardium, liver and the like can also occur. The death number of the model group is 5 (15 in total) within 1 week after the virus attack, the death number of the blank group is 1 (15 in total), and the success of model establishment can be primarily judged from clinical symptoms and the condition of section examination.
3.2 diarrhea score and mortality analysis Table
Note that: * Represents P < 0.05 compared to the blank, and represents P < 0.01 compared to the blank;
# represents P < 0.05 compared to the model group, # represents P < 0.01 compared to the blank group
From the above results, it can be seen that, through animal model verification, the effect of the embodiment 1 (fermented traditional Chinese medicine preparation group) and the traditional Chinese medicine mixed powder group of the invention on reducing diarrhea rate of the broiler chickens of the escherichia coli is superior to that of the commercial pulsatilla chinensis oral liquid, and mortality is obviously lower than that of the model group, so that the invention has obvious effect of improving diarrhea of the broiler chickens caused by the escherichia coli.
Example 8 clinical efficacy test
The invention has the effect test for preventing and treating the intestinal diseases of poultry
1. Purpose of test
Through clinical application of the preparation provided by the invention, whether the preparation provided by the invention can prevent or reduce intestinal diseases of chicken flocks is verified.
2. Test protocol
2.1 test subjects
The test selects chicken groups with similar ages in days and similar medication procedures (the number of chickens with the ages of 7 days is thirty-ten thousand). Randomly dividing into three groups, wherein one group is the group of the traditional Chinese medicine mixed powder without fermentation in the embodiment 1 of the invention, the other group is the commercial white head Weng Koufu liquid in the control group (according to the normal treatment mode in cultivation, the drug type and mode are recorded, and the effect comparison is carried out).
2.2 test materials
The samples of the test are provided by veterinary drug research and development centers of pharmaceutical research and development institute of the biological pharmaceutical industry, inc., and the experiment 1 group is 1000 g/bag of the fermented traditional Chinese medicine preparation in the embodiment 1 of the invention, and the batch number is 20191201. Experiment 2 is the traditional Chinese medicine mixed powder in the invention, and the control group is the commercial white head Weng Koufu liquid (which can be effectively used for treating diarrhea diseases).
2.3 test methods
The control group and the test group adopt consistent feeding standards and daily ration of a test chicken factory. Test group dosing method: the dosage of the drinking water administration method is as follows: in the experiment 1, the fermented traditional Chinese medicine preparation in the invention example 1 is used, 1000 jin of materials are mixed with 1000g (one bag) of the product, and the mixture is used for 1 week once a day; in the experiment 2, the traditional Chinese medicine mixed powder is used, 1000 jin of materials are mixed according to 1000g (one bag) of the traditional Chinese medicine mixed powder, and the mixed powder is used for 1 week once a day; the control group is commercial white head Weng Koufu liquid, and is added with water 800 jin per 500ml, once daily for 1 week.
| Group of | Number of chickens (Only) | Diarrhea rate | Cure rate | Mortality rate of |
| Experiment 1 group | 10000 | 8.2％ | 75％ | 2％ |
| Experiment 2 group | 10000 | 12.6％ | 65％ | 4％ |
| Control group | 10000 | 22.5％ | 10％ | 10％ |
3 test results
From the results, clinical verification proves that the effects of the traditional Chinese medicine fermentation group and the traditional Chinese medicine mixed powder on reducing the diarrhea rate of the broiler chickens are superior to those of the commercial pulsatilla chinensis bunge oral liquid, and the mortality rate is obviously lower than that of a control group. In particular, the traditional Chinese medicine fermentation group has more remarkable effect. Therefore, the product of the invention can effectively prevent and treat the occurrence of intestinal diseases of poultry.
The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (6)
1. A fermented Chinese medicinal preparation for preventing and treating diarrhea of fowl is prepared by fermenting substrate and fermentation brothThe fermentation substrate is characterized by comprising the following components in percentage by weight: 20% of bran, 20% of bean pulp and 60% of traditional Chinese medicine mixed powder; the addition amount of the fermentation bacteria liquid is 2.5 percent of the total amount of fermentation substrates, and the fermentation bacteria liquid contains lactobacillus plantarum 1 multiplied by 10 8～9 cfu/mL, bacillus subtilis 1×10 8～9 cfu/mL, microzyme 1×10 8～9 cfu/mL, wherein the lactobacillus plantarum is lactobacillus plantarum RP1 and is preserved in China center for type culture Collection, the preservation address is university of Wuhan, and the preservation number is CCTCCNO: m2018802;
wherein, each traditional Chinese medicine component of the traditional Chinese medicine mixed powder is as follows: cortex phellodendri, dark plum, poplar flower, purslane, dried ginger, bighead atractylodes rhizome, magnolia officinalis, medicated leaven, hawthorn, myrobalan and astragalus, and the traditional Chinese medicine mixed powder comprises the following traditional Chinese medicine components in parts by weight: 30 parts of phellodendron, 20 parts of dark plum, 70 parts of poplar flower, 30 parts of purslane, 20 parts of dried ginger, 40 parts of bighead atractylodes rhizome, 40 parts of magnolia officinalis, 70 parts of medicated leaven, 20 parts of hawthorn, 20 parts of myrobalan and 40 parts of astragalus membranaceus.
2. A method for preparing a fermented traditional Chinese medicine preparation for preventing and treating diarrhea of poultry according to claim 1, which comprises the following specific steps:
(1) Preparing fermentation bacteria liquid: inoculating the strains into special culture mediums respectively, culturing seed solutions according to the special conditions, and stopping culturing when the strains grow to the logarithmic growth phase and are respectively transferred into a fermentation tank for expansion culture and reach the logarithmic growth phase;
(2) The preparation of the traditional Chinese medicine mixed powder comprises the following steps: removing impurities from the medicinal materials respectively, crushing the medicinal materials to 80-100 meshes, and mixing the medicinal materials according to the weight parts;
(3) Removing impurities from bran and bean pulp, pulverizing in a pulverizer, taking out, placing in a mixer, and adding the prepared traditional Chinese medicine mixed powder into the mixer for fully and uniformly mixing;
(4) Spraying the fermentation broth into a mixer for mixing, adding a certain amount of water, and fermenting after fully and uniformly mixing.
3. The method according to claim 2, wherein the fermentation method in the step (4) is: after the fermentation bacteria liquid and the water are added and fully mixed, the materials are transferred to a discharging machine, and after being accurately weighed, the materials are filled in a fermentation bag with a one-way breather valve and sealed, and are placed in a fermentation chamber for fermentation.
4. A method according to claim 3, wherein the conditions under which fermentation is carried out in the fermentation chamber are: and fermenting at room temperature of 25-37 deg.c and humidity of 50-70% for 7-14 days.
5. The method according to claim 4, wherein the conditions for fermentation in the fermentation chamber are: the room temperature is 28 ℃, the humidity is 65 percent, and the fermentation is carried out for 12 days.
6. The method according to claim 2, wherein the amount of the added water in the step (4) is controlled to 40 to 60% of the total water content.
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| CN110946907A (en) * | 2019-12-25 | 2020-04-03 | 六安恒佳生物科技有限公司 | Traditional Chinese medicine composition for young poultry, liquid fermented traditional Chinese medicine preparation and preparation method thereof |
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| CN110946907A (en) * | 2019-12-25 | 2020-04-03 | 六安恒佳生物科技有限公司 | Traditional Chinese medicine composition for young poultry, liquid fermented traditional Chinese medicine preparation and preparation method thereof |
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| Title |
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| 自拟中药乌梅散超微粉治疗鸭腹泻试验观察;张丁华等;《中兽医医药杂志》;20151231(第5期);第48-50页 * |
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