CN111166880A - Preparation method of egg yolk antibody effervescent tablets for egg drop syndrome - Google Patents
Preparation method of egg yolk antibody effervescent tablets for egg drop syndrome Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
- A61K9/0007—Effervescent
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/20—Pills, tablets, discs, rods
- A61K9/2004—Excipients; Inactive ingredients
- A61K9/2022—Organic macromolecular compounds
- A61K9/2063—Proteins, e.g. gelatin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/20—Pills, tablets, discs, rods
- A61K9/2004—Excipients; Inactive ingredients
- A61K9/2068—Compounds of unknown constitution, e.g. material from plants or animals
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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
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
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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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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/11—Immunoglobulins specific features characterized by their source of isolation or production isolated from eggs
Abstract
The invention provides a preparation method of a yolk antibody effervescent tablet for egg drop syndrome, which comprises the steps of immunizing healthy high-yield laying hens by using an egg drop syndrome vaccine, collecting hyperimmune eggs, and extracting a yolk antibody; is prepared by adding probiotics and freeze-drying protective agent and effervescent material. The method creatively designs a new formulation of the egg-loss syndrome egg yolk antibody, enriches new ways for preventing and controlling the egg-loss syndrome, has low cost, high antibody titer, convenient use and high biological safety, and effectively ensures the treatment effect of the obtained egg yolk antibody.
Description
Technical Field
The invention belongs to the technical field of biological products for livestock, and particularly relates to a preparation method of a yolk antibody effervescent tablet for egg drop syndrome.
Background
The egg drop syndrome of the chicken is the egg drop syndrome of the chicken, the disease is an infectious disease which is caused by the egg drop syndrome virus and is characterized by egg drop or special egg laying, the infectious disease frequently occurs in a large-scale intensive chicken farm and has extremely high propagation speed, the disease is mainly infected by egg transmission, and the diseases are susceptible to quails, chickens, turkeys, ducks, geese, guinea fowls, sparrows and the like. The sick chicken is the main infection source of the disease, the susceptibility of different varieties of chicken is slightly different, the chicken laying brown shell eggs is most susceptible, the prevalence rate of the chicken laying white shell eggs is low, the disease is not obvious seasonal, the disease can occur all the year round, the reproductive system of the laying hens aged 26-34 weeks is mainly invaded, the eggshell formation mechanism of egg yolk discharge is disordered, the egg laying rate is reduced, and special-shaped eggs without shells or soft shells appear.
Eggs are an important food protein source on the human dining table, so that laying hen breeding is an important support for national dining table economy. At present, the prevention and control means of egg drop syndrome mainly depend on vaccines, but in recent years, the vaccine prevention and control effect is greatly reduced due to the fact that the food consumption of the vaccines is reduced in the empty window period and the stress is large.
Disclosure of Invention
The invention aims to provide a yolk antibody effervescent tablet for egg drop syndrome of chicken, which has novel design and solves the embarrassing situation of vaccine morbidity in the empty window period in the prior art; meanwhile, the composition has the effects of prevention and treatment, and avoids the problems of large stress, poor effect and the like.
In order to achieve the purpose, the invention is realized by the following technical scheme:
1. a preparation method of a yolk antibody effervescent tablet for egg drop syndrome of chicken comprises the following steps:
(1) taking non-immunized 23-week-old healthy laying hens, performing 3 times of immunization injection of the egg drop syndrome vaccine, wherein the immunization interval is 10 days, the dosage is 1 ml/egg, and collecting hyperimmune eggs when the serum antibody titer is detected to be 1: 512;
(2) cleaning and disinfecting the hyperimmune egg obtained in the step (1), collecting and stirring to obtain egg yolk homogenate;
(3) adding 1.5 times volume of physiological saline and 0.1% caprylic acid into the egg yolk homogenate obtained in the step (2), stirring for 0.5-1.5 hours while adding, standing at room temperature for 2-6 hours, centrifuging for 30 minutes at the rotating speed of 4200-5500 r/min, discarding the precipitate, adding 1.2% sulfate dextran into the supernatant, stirring for 10-20 minutes, filtering the supernatant by a cylindrical filter element with the aperture of 5 mu m and 1 mu m, concentrating by a hollow fiber ultrafiltration column with the cut-off molecular weight of 10kD for 3-4 times, adjusting the pH value to 6.8-7.0 by NaOH, and filtering and sterilizing by a cylindrical filter element with the aperture of 0.45 mu m and 0.22 mu m to obtain the filtrate with the antibody titer of 1: 256.
(4) Preparing probiotic liquid: respectively culturing the bifidobacterium lactis and the bacillus subtilis, and mixing the two bacterium liquids to obtain a probiotic liquid;
(5) uniformly mixing the filtrate obtained in the step (3) and the probiotic liquid obtained in the step (4) in a volume ratio of 1:1 under an aseptic condition, adding a freeze-drying protective agent in a volume ratio of 1:1, uniformly mixing, and carrying out freeze-vacuum drying treatment to obtain freeze-dried powder;
(6) and (4) uniformly mixing the freeze-dried powder obtained in the step (5) with an effervescent material, tabletting, and subpackaging to obtain the egg yolk antibody effervescent tablet for egg drop syndrome.
Moreover, the method for sterilizing the high-immunity eggs and collecting the yolk in the step (2) comprises the following steps: washing the high-immunity eggs obtained in the step 1) with clear water, soaking the eggs into 0.1% benzalkonium bromide water solution for disinfection for 15 minutes, taking out the eggs, airing the eggs, disinfecting the eggs with ethylene oxide aerosol for 30 minutes, feeding the eggs into an egg white and egg yolk separator for separation to obtain egg yolk, and stirring the egg yolk to obtain egg yolk homogenate.
And in the step (4), the bifidobacterium lactis and the bacillus subtilis are cultured according to the following steps: preheating an MRS liquid culture medium to 37 ℃, simultaneously inoculating bifidobacterium lactis seed liquid according to the inoculation amount of 1%, standing and culturing for 18-20 hours at 37 ℃, and harvesting the bacterial liquid to 2-8 ℃ for later use; preheating a bacillus culture medium to 37 ℃, simultaneously inoculating bacillus subtilis seed liquid according to the inoculation amount of 2%, standing and culturing for 16-20 hours at 37 ℃, and harvesting the bacterial liquid to 2-8 ℃ for storage and later use.
And the freeze-drying protective agent in the step (5) contains 10-30 parts of astragalus polysaccharide, 2-5 parts of gelatin, 5-10 parts of cane sugar and 5-20 parts of dandelion flavone in each 1000ml of phosphate buffer.
Furthermore, the preparation method of the effervescent material in the step (6) comprises the following steps:
weighing 22 parts of citric acid, 15 parts of cane sugar, 35 parts of sodium bicarbonate, 10 parts of sodium carboxymethyl starch, 1 part of sweet orange powder, 4 parts of 10% PVP absolute ethyl alcohol, 01 parts of PEG60001 and 0.5 part of magnesium stearate according to parts by weight, fully mixing the components, drying at 40-65 ℃, sieving with a 6-10 mesh sieve to prepare granules for later use, and performing Co-60 radiation sterilization to prepare the effervescent material.
The invention has the advantages and beneficial effects that:
1. the invention reduces the requirements of cold chain transportation and storage of products and increases the convenience for farmers to use.
2. The egg yolk antibody provided by the invention has the effects of a microecological preparation and the egg drop syndrome egg yolk antibody when being fed by drinking water, so that manpower and material resources are reduced, and the stress to chicken flocks is reduced.
3. The invention can enhance the immunity of chicken flocks, improve the overall immunity effect of the chicken flocks and reduce the risk of mixed infection diseases of the chicken flocks.
Detailed Description
The invention is further illustrated by the following examples. It should be noted that: the following examples are illustrative and not intended to be limiting, and are not intended to limit the scope of the invention.
The egg drop syndrome vaccine related by the invention can be purchased in the market, such as astragalus polysaccharide, dandelion flavone, sucrose, alum, bifidobacterium lactis and bacillus subtilis.
Example 1
A preparation method of a yolk antibody effervescent tablet for egg drop syndrome of chicken comprises the following steps:
(1) taking non-immunized 23-week-old healthy laying hens, performing 3 times of immunization injection on inactivated vaccine of infectious bursal disease oil emulsion, wherein the immunization interval is 10 days, the dosage is 1 ml/egg, and collecting hyperimmune eggs when the serum antibody titer is detected to be 1: 512;
(2) taking the high-immunity eggs obtained in the step (1), washing the high-immunity eggs by using clear water, soaking the high-immunity eggs into 0.1% benzalkonium bromide water solution for disinfection for 15 minutes, taking out the high-immunity eggs, airing the high-immunity eggs, disinfecting the high-immunity eggs by using ethylene oxide aerial fog for 30 minutes, sending the high-immunity eggs into an egg white and egg yolk separator for separation to obtain egg yolks, and stirring the egg yolk;
(3) adding 1.5 times volume of physiological saline and 0.1% n-octanoic acid into the egg yolk homogenate obtained in the step (2), stirring for 0.5 hour while adding, standing at room temperature for 2 hours, centrifuging at 4200r/min for 30 minutes, discarding the precipitate, adding 1.2% sulfate dextran into the supernatant, stirring for 10 minutes, filtering the supernatant with a cylindrical filter element with a pore size of 5 μm and a cylinder with a pore size of 1 μm, concentrating by a hollow fiber ultrafiltration column with a molecular weight cut-off of 10kD for 3 times, adjusting the pH value to 6.8 with NaOH, and filtering and sterilizing with a cylindrical filter element with a pore size of 0.45 μm and a cylinder with a pore size of 0.22 μm to obtain a filtrate with an antibody titer of 1: 256.
(4) Preparing probiotic liquid: preheating a lactobacillus liquid culture medium to 37 ℃, simultaneously inoculating bifidobacterium lactis seed liquid according to 1 percent of inoculation amount, standing and culturing for 18 hours at 37 ℃, and harvesting the bacterial liquid to 2-8 ℃ for later use; preheating a bacillus culture medium to 37 ℃, simultaneously inoculating bacillus subtilis seed liquid according to the inoculation amount of 2%, standing and culturing for 16 hours at 37 ℃, and harvesting the bacterial liquid to 2-8 ℃ for storage and later use.
(5) Preparing a freeze-drying protective agent: adding 10g of astragalus polysaccharide, 5g of dandelion flavone, 2g of gelatin and 5g of sucrose into each 1000ml of phosphate buffer to prepare a freeze-drying protective agent, uniformly mixing the filtrate obtained in the step (3) and the probiotic liquid obtained in the step (4) in a volume ratio of 1:1 under an aseptic condition, adding the freeze-drying protective agent in a volume ratio of 1:1, uniformly mixing, and carrying out freeze-drying treatment in vacuum to obtain freeze-dried powder;
(6) preparing an effervescent material: 22g of citric acid, 15g of cane sugar, 35g of sodium bicarbonate, 10g of sodium carboxymethyl starch, 1g of sweet orange powder, 4g of 10% PVP absolute ethyl alcohol, 4g of PEG60001g and 0.5g of magnesium stearate are weighed according to parts by weight, fully mixed, dried at 40 ℃ and sieved by a 6-mesh sieve to prepare granules for later use, and the granules are sterilized by Co-60 radiation to prepare the effervescent material.
(7) And (4) uniformly mixing the freeze-dried powder obtained in the step (5) with an effervescent material, tabletting, and subpackaging to obtain the egg drop syndrome egg yolk antibody effervescent tablets.
Example 2
A preparation method of a probiotics-free egg yolk antibody effervescent tablet for egg drop syndrome of chickens comprises the following steps:
(1) taking non-immunized 23-week-old healthy laying hens, performing 3 times of immunization injection of the egg drop syndrome vaccine, wherein the immunization interval is 10 days, the dosage is 1 ml/egg, and collecting hyperimmune eggs when the serum antibody titer is detected to be 1: 256;
(2) taking the high-immunity eggs obtained in the step (1), washing the high-immunity eggs by using clear water, soaking the high-immunity eggs into 0.1% benzalkonium bromide water solution for disinfection for 15 minutes, taking out the high-immunity eggs, airing the high-immunity eggs, disinfecting the high-immunity eggs by using ethylene oxide aerial fog for 30 minutes, sending the high-immunity eggs into an egg white and egg yolk separator for separation to obtain egg yolks, and stirring the egg yolk;
(3) adding 1.5 times volume of physiological saline and 0.1% n-caprylic acid into the egg yolk homogenate obtained in the step (2), stirring for 1.5 hours while adding, standing at room temperature for 6 hours, centrifuging for 30 minutes at the rotating speed of 5500r/min, discarding the precipitate, adding 1.2% sulfate dextran into the supernatant, stirring for 20 minutes, filtering the supernatant by a cylindrical filter element with the aperture of 5 mu m and the aperture of 1 mu m, concentrating by a hollow fiber ultrafiltration column with the molecular weight cutoff of 10kD for 3-4 times, adjusting the pH value to 7.0 by NaOH, and filtering and sterilizing by a cylindrical filter element with the molecular weight of 0.45 mu m and the 0.22 mu m to obtain the filtrate with the antibody titer of 1: 128.
(4) Preparing a freeze-drying protective agent: adding 30g of astragalus polysaccharide, 20g of dandelion flavone, 5g of gelatin and 10g of sucrose into each 1000ml of phosphate buffer solution to prepare a freeze-drying protective agent, adding the freeze-drying protective agent into the filtrate obtained in the step (3) according to the volume ratio of 1:1, uniformly mixing, and carrying out freeze-vacuum drying treatment to obtain freeze-dried powder;
(5) preparing an effervescent material: weighing 22g of citric acid, 15g of cane sugar, 35g of sodium bicarbonate, 10g of sodium carboxymethyl starch, 1 part of sweet orange powder, 4g of 10% PVP absolute ethyl alcohol, PEG60001g and 0.5g of magnesium stearate according to weight, fully mixing the above components, drying at 65 ℃, sieving with a 10-mesh sieve to obtain granules for later use, and performing Co-60 radiation sterilization to obtain the effervescent material.
(6) And (4) uniformly mixing the freeze-dried powder obtained in the step (4) with an effervescent material, tabletting and subpackaging to obtain the egg yolk antibody effervescent tablet without probiotics for egg drop syndrome.
Example 3
A preparation method of egg drop syndrome egg yolk antibody effervescent tablets without freeze-drying protective agents comprises the following steps:
(1) taking non-immunized 23-week-old healthy laying hens, performing 3 times of immunization injection of the egg drop syndrome vaccine, wherein the immunization interval is 10 days, the dosage is 1 ml/egg, and collecting hyperimmune eggs when the serum antibody titer is detected to be 1: 256;
(2) taking the high-immunity eggs obtained in the step (1), washing the high-immunity eggs by using clear water, soaking the high-immunity eggs into 0.1% benzalkonium bromide water solution for disinfection for 15 minutes, taking out the high-immunity eggs, airing the high-immunity eggs, disinfecting the high-immunity eggs by using ethylene oxide aerial fog for 30 minutes, sending the high-immunity eggs into an egg white and egg yolk separator for separation to obtain egg yolks, and stirring the egg yolk;
(3) adding 1.5 times volume of physiological saline and 0.1% n-caprylic acid into the egg yolk homogenate obtained in the step (2), stirring for 1.5 hours while adding, standing at room temperature for 6 hours, centrifuging for 30 minutes at the rotating speed of 5500r/min, discarding the precipitate, adding 1.2% sulfate dextran into the supernatant, stirring for 20 minutes, filtering the supernatant by a cylindrical filter element with the aperture of 5 mu m and the aperture of 1 mu m, concentrating by a hollow fiber ultrafiltration column with the molecular weight cutoff of 10kD for 3-4 times, adjusting the pH value to 7.0 by NaOH, and filtering and sterilizing by a cylindrical filter element with the molecular weight of 0.45 mu m and the 0.22 mu m to obtain the filtrate with the antibody titer of 1: 128.
(4) Preparing probiotic liquid: preheating a lactic acid bacteria liquid culture medium to 37 ℃, simultaneously inoculating bifidobacterium bifidum liquid according to 1 percent of inoculation amount, standing and culturing for 20 hours at 37 ℃, and harvesting the bacterial liquid to 2-8 ℃ for later use; preheating a bacillus culture medium to 37 ℃, simultaneously inoculating bacillus subtilis seed liquid according to the inoculation amount of 2%, standing and culturing for 20 hours at 37 ℃, and harvesting the bacterial liquid to 2-8 ℃ for later use.
(5) Uniformly mixing the filtrate obtained in the step (3) and the probiotic liquid obtained in the step (4) in a volume ratio of 1:1 under an aseptic condition, and carrying out freeze vacuum drying treatment to obtain dry powder;
(6) preparing an effervescent material: weighing 22g of citric acid, 15g of cane sugar, 35g of sodium bicarbonate, 10g of sodium carboxymethyl starch, 1g of sweet orange powder, 4g of 10% PVP absolute ethyl alcohol, PEG60001g and 0.5g of magnesium stearate according to weight, fully mixing the above components, drying at 65 ℃, sieving with a 10-mesh sieve to obtain granules for later use, and performing Co-60 radiation sterilization to obtain the effervescent material.
(7) And (4) uniformly mixing the dry powder obtained in the step (5) with an effervescent material, tabletting and subpackaging to obtain the egg drop syndrome egg yolk antibody effervescent tablet without the freeze-drying protective agent.
Clinical trial
1. Test materials:
the egg-drop syndrome egg yolk antibody effervescent tablet is prepared according to the example 1 and is used for the group 1;
preparing a probiotic-free egg drop syndrome egg yolk antibody effervescent tablet according to example 2; for group 2;
preparing a yolk antibody effervescent tablet of egg drop syndrome of the chicken without a freeze-drying protective agent according to the example 3; for 3 groups;
the general antibody and the chemical drug group are all commercial products and are used according to the instructions.
2. The use method of the invention comprises the following steps: group 1: the dosage is calculated according to the fact that 1 tablet of the invention is dissolved in 1kg of water, and the invention is freely drunk by poultry; the dosage of the 2 groups and the 3 groups is calculated according to 1 tablet dissolved in 1kg of water, and the invention is freely drunk by poultry.
3. Test grouping and processing
In 2019, egg drop syndrome occurs in a certain chicken farm in a Shandong Weifang, the egg yield is suddenly reduced, 50 feathers of sick white legging chickens of 3 weeks old are selected and randomly divided into 5 groups, each group comprises 10 chickens, 1-3 groups are test groups, 4 groups are chicken egg drop syndrome common antibody treatment groups, 5 groups are blank control groups, all the groups are fed with complete feed under the same feeding environment, 10 days are observed, and the test results are as shown in the following table.
Table effect test of the present invention for treating egg drop syndrome
Conclusion
The cure rate of the egg-yolk antibody effervescent tablet for egg drop syndrome of chicken is 100%, which is obviously superior to other groups; the egg yolk antibody effervescent tablets for the egg drop syndrome of the chickens, which do not contain probiotics, and the egg yolk antibody effervescent tablets for the egg drop syndrome of the chickens, which do not contain freeze-drying protective agents, have obvious antibody effects, but have low feed utilization rate and slightly reduced egg laying quality due to the influence on feed intake and physique before and after the immunization period, but still have better curative effect than the commercial ordinary antibody group, so that the effect of treating the egg drop syndrome of the chickens is obvious.
Claims (5)
1. A preparation method of a yolk antibody effervescent tablet for egg drop syndrome is characterized by comprising the following steps: the preparation method comprises the following steps:
(1) taking non-immunized 23-week-old healthy laying hens, performing 3 times of immunization injection of the egg drop syndrome vaccine, wherein the immunization interval is 10 days, the dosage is 1 ml/egg, and collecting hyperimmune eggs when the serum antibody titer is detected to be 1: 512;
(2) cleaning and disinfecting the hyperimmune egg obtained in the step (1), collecting and stirring to obtain egg yolk homogenate;
(3) adding 1.5 times volume of physiological saline and 0.1% caprylic acid into the egg yolk homogenate obtained in the step (2), stirring for 0.5-1.5 hours while adding, standing at room temperature for 2-6 hours, centrifuging for 30 minutes at the rotating speed of 4200-5500 r/min, discarding the precipitate, adding 1.2% sulfate dextran into the supernatant, stirring for 10-20 minutes, filtering the supernatant by a cylindrical filter element with the aperture of 5 mu m and 1 mu m, concentrating by a hollow fiber ultrafiltration column with the cut-off molecular weight of 10kD for 3-4 times, adjusting the pH value to 6.8-7.0 by NaOH, and filtering and sterilizing by a cylindrical filter element with the aperture of 0.45 mu m and 0.22 mu m to obtain the filtrate with the antibody titer of 1: 256.
(4) Preparing probiotic liquid: respectively culturing the bifidobacterium lactis and the bacillus subtilis, and mixing the two bacterium liquids to obtain a probiotic liquid;
(5) uniformly mixing the filtrate obtained in the step (3) and the probiotic liquid obtained in the step (4) in a volume ratio of 1:1 under an aseptic condition, adding a freeze-drying protective agent in a volume ratio of 1:1, uniformly mixing, and carrying out freeze-vacuum drying treatment to obtain freeze-dried powder;
(6) and (4) uniformly mixing the freeze-dried powder obtained in the step (5) with an effervescent material, tabletting, and subpackaging to obtain the egg yolk antibody effervescent tablet for egg drop syndrome.
2. The preparation method of the yolk antibody effervescent tablet for egg drop syndrome of chicken as claimed in claim 1, wherein the method for sterilizing the high-immunity egg and collecting yolk in step (2) is as follows: washing the high-immunity eggs obtained in the step 1) with clear water, soaking the eggs into 0.1% benzalkonium bromide water solution for disinfection for 15 minutes, taking out the eggs, airing the eggs, disinfecting the eggs with ethylene oxide aerosol for 30 minutes, feeding the eggs into an egg white and egg yolk separator for separation to obtain egg yolk, and stirring the egg yolk to obtain egg yolk homogenate.
3. The preparation method of the egg yolk antibody effervescent tablet for egg drop syndrome of chicken as claimed in claim 1, wherein the bifidobacterium lactis and the bacillus subtilis are cultured according to the following steps in the step (4) respectively: preheating an MRS liquid culture medium to 37 ℃, simultaneously inoculating bifidobacterium lactis seed liquid according to the inoculation amount of 1%, standing and culturing for 18-20 hours at 37 ℃, and harvesting the bacterial liquid to 2-8 ℃ for later use; preheating a bacillus culture medium to 37 ℃, simultaneously inoculating bacillus subtilis seed liquid according to the inoculation amount of 2%, standing and culturing for 16-20 hours at 37 ℃, and harvesting the bacterial liquid to 2-8 ℃ for storage and later use.
4. The method for preparing the yolk antibody effervescent tablet for egg drop syndrome of chicken as claimed in claim 1, wherein the lyoprotectant in step (5) comprises 10-30 parts of astragalus polysaccharides, 2-5 parts of gelatin, 5-10 parts of sucrose, and 5-20 parts of taraxylflavone per 1000ml of phosphate buffer.
5. The preparation method of the yolk antibody effervescent tablet for egg drop syndrome of chicken according to claim 1, characterized in that the preparation method of the effervescent material in step (6) comprises the following steps:
weighing 22 parts of citric acid, 15 parts of cane sugar, 35 parts of sodium bicarbonate, 10 parts of sodium carboxymethyl starch, 1 part of sweet orange powder, 4 parts of 10% PVP absolute ethyl alcohol, 01 parts of PEG60001 and 0.5 part of magnesium stearate according to parts by weight, fully mixing the components, drying at 40-65 ℃, sieving with a 6-10 mesh sieve to prepare granules for later use, and performing Co-60 radiation sterilization to prepare the effervescent material.
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| CN202010000048.8A CN111166880A (en) | 2020-01-01 | 2020-01-01 | Preparation method of egg yolk antibody effervescent tablets for egg drop syndrome |
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| CN202010000048.8A CN111166880A (en) | 2020-01-01 | 2020-01-01 | Preparation method of egg yolk antibody effervescent tablets for egg drop syndrome |
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Application publication date: 20200519 |