CN103550770B - Preparation method of compound effervescent tablets for treating chicken infectious bursal disease - Google Patents
Preparation method of compound effervescent tablets for treating chicken infectious bursal disease Download PDFInfo
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Abstract
The invention discloses a preparation method of compound effervescent tablets for treating chicken infectious bursal disease, which belongs to the technical field of veterinary biological products. The preparation method comprises the following steps: carrying out passage and culture on a vaccine cell; reproducing a cell virus seed; reproducing a vaccine virus liquid; preparing a probiotics mixing bacteria liquid; freeze-drying a mixed preparation; and preparing the effervescent tablets. The preparation method has the beneficial effects that the requirement on transportation of the compound vaccine effervescent tablets for treating chicken infection bursal disease prepared by the invention is lowered and the convenience of users is improved; the compound vaccine effervescent tablets are capable of feeding microecologics while immunizing a chicken infectious bursal disease live vaccine, so that manpower and material resources are reduced and the stress of chicken flocks is reduced; the compound vaccine effervescent tablets are capable of improving the immunity of the chicken flocks, enhancing the overall immune effects of the chicken flocks and reducing the antibody dispersion of the chicken flocks, so that the antibody of the chicken flocks is kept on the same level and the infection risk of the chicken flocks is reduced.
Description
Technical field
The invention belongs to veterinary biologics technical field, be specifically related to a kind of preparation method of infectious bursal disease compound effervescent tablet.
Background technology
Along with the rise of intensive aviculture and the increase of international trade, fowl diseases is popular to be on the rise, and causes the high mortality of poultry, seriously governs the development of poultry husbandry, causes the loss that world's poultry husbandry is serious, jeopardizes economic development, food safety and social stability.Infectious bursal disease (Infection bursal disease, IBD) be by infectious bursa of Fabricius virus (Infection bursal disease virus, a kind of high degree in contact sexually transmitted disease of the chicken IBDV) caused and turkey, main infection 3-12 week age young chicken, this virus is mainly in the lymphocyte internal breeding of fabricius bursa, and damage is in various degree caused to other immune organ, finally causes the fabricius bursa atrophy of chicken and cause the immunosuppressant of chicken.This disease is without significantly seasonal, all can occur throughout the year, propagate usually through approach such as contaminated feedstuff, drinking-water, feces, hen house apparatus, staff garments, incubation period is 1-5 days, and after there is symptom the 3rd day starts dead, within 4-6 days, be peak mortality phases, stop death gradually later, sickness rate can reach more than 90%, and mortality rate not etc., is not generally 10%-50%.Primary disease can cause losing weight clinically, poor growth and the symptom such as skeletal muscle is hemorrhage, often occur together when sanitary condition is poor other diseases, mortality rate improves further, and chickling individuality is little, and premunition is poor, once generation epidemic disease, then infect fast, mortality rate is high, and mortality rate reaches more than 80%, loss is larger, has carried out huge economic loss to poultry industrial belt.
Vaccine is the goods be made up after breeding and process of the good pathogenic microorganism of immunogenicity, after inoculation animal body, body is stimulated to produce specific antibody, after the antibody titer in body reaches certain numerical value, just can resist invasion and attack, the infection of this pathogenic microorganism, play the effect preventing this disease.Along with the fast development of China's aviculture, more and more important to the control of infectious bursal disease.
Polysaccharide is one of active ingredient of Chinese herbs, and large quantity research shows, polysaccharide and saccharide complex are not only in vivo as energy resource and constituent material, the more important thing is that it is present in all membrane structures, participates in the various activities of cell in biosis.Polysaccharose substance is the important component part of all Living organisms, has scavenging free radicals, improves the ability of activities of antioxidant enzymes and anti-lipid peroxidation.
Vitamin is requisite organic compound in organism metabolism.The chemical plant that body is very complicated just as one, constantly carries out various biochemical reaction.Its reaction has substantial connection with the catalytic action of enzyme.Enzyme will produce activity, and coenzyme must be had to participate in.Known many vitamin are the coenzyme of enzyme or the ingredient of coenzyme.Therefore, vitamin is the important substance maintaining and regulate body homergy.Vitamin is one of indispensable nutrient substance in growth of animal process, and especially when occurring, the effect of vitamin is more obvious.Suitable edible vitamin can reduce Animal stress, reduces disease incident, improves animal disease resistant ability.
Flavone compound is the polyatomic phenol material that occurring in nature exists, and is also one of main active in nature medicinal plants.It refers to have 15 carbon atoms and with the three ring natural organic matters that the mode of C6-C3-C6 is formed, be the secondary metabolism product that plant produces in long-term natural selection, now isolation identification have kind more than 4000.It is extensively present in fruit and vegerable, Chinese herbal medicine, have no side effect, it has the medicine healthy sofa functions such as significant scavenger interior free yl, aging resistance, mutation, Adjust-blood lipid blood pressure lowering, it is the natural organic oxidation-resistant agent that a class has DEVELOPMENT PROSPECT, these anti-oxidation active substances can reduce and scavenger interior free yl, have prophylactic effect.
Probiotic bacteria has prebiotic effect to people and animals.Probiotic bacteria in microbial ecological agent can produce antibiotic substance, and competes the Ecological niche with noxious bacteria, suppresses the growth and breeding of pathogenic bacterium, the colony balance of adjustment and recovery intestinal.In addition, microbial ecological agent can become the nonspecific immunity factor, and by antibacterial itself or cell wall constituent stimulation of host cell, activating immune system, improves body antibody horizontal or macrophage activity, thus enhancing body resistance.Profitable strain in microbial ecological agent is multiplied at digestive tract, can in facilitating digestion road a series of nutritional labeling such as several amino acids, vitamin effective synthesis and absorb, thus promote animal growth and development and weightening finish.Simultaneously in environmental protection, fecaluria stench can be removed, purifying ecological environment.Probiotic bacteria in microbial ecological agent decomposes and synthesis by carrying out fermentation to harmful substance, effectively reduces the content of poisonous and harmful substance.
At present, commercially available microbial ecological agent is mostly lyophilized powder, and can directly mix in feedstuff, oral administration mode be absorbed by animal body.Although this feed mode is simple and convenient, because microbial ecological agent is mostly lactobacillus, its vitality is comparatively fragile, mixes and significantly can reduce viable count in feedstuff.In addition, because same feedstuff is fed jointly, be difficult to the accurate-metering ensureing that animal absorbs, thus affect the treatment.The product of lyophilised state all will remain on " cold chain " environment under lower state of temperature in the whole storage used and transportation in addition, and this brings inconvenience to the storage of product and the use of transport and user.
Summary of the invention
For prior art Problems existing, the object of the invention is to design the technical scheme of the preparation method that a kind of infectious bursal disease compound effervescent tablet is provided.
The preparation method of described a kind of infectious bursal disease compound effervescent tablet, is characterized in that comprising following processing step:
1) the going down to posterity and cultivation of seedling cell: get chick embryo fibroblast continuous cell line DF-1 cell line through pancreatin cell dispersal liquid had digestive transfer culture, continuing to cultivate with cell growth medium, when forming good monolayer, going down to posterity or virus inoculation for continuing in culture bottle;
2) breeding of cell seed culture of viruses: get production infectious bursa of Fabricius virus B87 strain, being inoculated in by 1% of maintenance medium volume grows up in the chick embryo fibroblast passage cell of good monolayer, put 37 ~ 38 DEG C to continue to cultivate in maintenance medium, harvesting liquid when cytopathy variability reaches more than 75%;
3) breeding of seedling venom: get the cell line culture bottle having formed good monolayer in step 1), discard cell growth medium, inoculation step 2) obtain containing the maintenance medium of the cell adapted malicious B87 strain of 1% fabricius bursa, put 37 ~ 38 DEG C to continue to cultivate, venom is gathered in the crops when cytopathy reaches more than 75%, less than-15 DEG C preservations, obtain infections chicken cloacal bursa virus suspension;
4) preparation of probiotic bacteria mixed bacteria liquid: extracting lactic acid bacterium and bacillus cereus are cultivated respectively, then will the lactic acid bacterial liquid and the mixing of bacillus cereus bacterium liquid that obtain be cultivated, and adjust bacterium number, obtain probiotic bacteria mixed bacteria liquid;
5) lyophilizing of mix preparation: the probiotic bacteria mixed bacteria liquid that the infections chicken cloacal bursa virus suspension and the step 4) that step 3) are obtained obtain aseptically by volume 1:1 mix homogeneously, 1:1 adds freeze drying protectant mix homogeneously by volume again, through lyophilisation after subpackage, obtain mix preparation lyophilized powder;
6) effervescent tablet preparation: mix preparation lyophilized powder step 5) obtained is mixed homogeneously with effervescent materials, tabletting, and subpackage obtains infectious bursal disease compound effervescent tablet.
The preparation method of described a kind of infectious bursal disease compound effervescent tablet, to it is characterized in that in described step 4) lactobacillus be bifidobacterium bifidum (
bifidobacterium bifidum), lactobacillus lactis (
lactobacillus lactis), Lactobacillus buchneri (
lactobacillus buchneri), bifidobacterium longum (
bifidobacterium longum) in one or more, described bacillus cereus be Bacillus licheniformis (
bacillus licheniformis), Bacillus coagulans (
bacillus coagulans), bacillus subtilis (
bacillus subtilis) in one or more.
The preparation method of described a kind of infectious bursal disease compound effervescent tablet, it is characterized in that in described step 4), lactobacillus and bacillus cereus are cultivated according to following steps respectively: MRS fluid medium is preheated to 37 DEG C, simultaneously by 2% inoculum concentration inoculating lactic acid bacterium seed liquor, 37 DEG C of quiescent culture 18 ~ 24 hours, results bacterium liquid to 4 DEG C saves backup; Bacillus culture medium is preheated to 37 DEG C, simultaneously by 1% inoculum concentration inoculation bacillus cereus seed liquor, 37 DEG C of quiescent culture 18 ~ 24 hours, results bacterium liquid to 4 DEG C saves backup.
The preparation method of described a kind of infectious bursal disease compound effervescent tablet, it is characterized in that in described step 5), freeze drying protectant is containing herbal polysaccharide 30 ~ 80g in every 1000ml phosphate buffer, Chinese medicine flavone 20 ~ 50g, vitamin combination 6 ~ 10g, gelatin 4 ~ 6g and sucrose 40 ~ 70g, described herbal polysaccharide and Chinese medicine flavone extract and obtain from following Chinese medicine composition: Radix Codonopsis, Radix Ginseng, Fructus Lycii, Herba Andrographis, Radix Achyranthis Bidentatae, Poria, Folium Ginkgo, Radix Glycyrrhizae, the Radix Astragali, Herba Urticae Cannabinae, Herba Equiseti Arvinsis and Radix Salviae Miltiorrhizae, described vitamin is combined as vitamin C, vitamin E, vitamin D3, vitamin B1, vitamin K and vitamin B2.
The preparation method of described a kind of infectious bursal disease compound effervescent tablet, is characterized in that described freeze drying protectant is containing herbal polysaccharide 50 ~ 70g, Chinese medicine flavone 30 ~ 40g, vitamin combination 7 ~ 9g, gelatin 4.5 ~ 5.5g and sucrose 50 ~ 60g in every 1000ml phosphate buffer.
The preparation method of described a kind of infectious bursal disease compound effervescent tablet, is characterized in that in described vitamin combination containing vitamin C 20 ~ 40%, vitamin E 10 ~ 30%, vitamin D3 5 ~ 15%, vitamin B1 15 ~ 35%, vitamin K 10 ~ 20%, vitamin B2 5 ~ 20%.
The preparation method of described a kind of infectious bursal disease compound effervescent tablet, is characterized in that described herbal polysaccharide and Chinese medicine flavone are obtained by following steps:
A, take each raw material of Chinese medicine by Radix Codonopsis 10 ~ 20 parts, Radix Ginseng 5 ~ 20 parts, Fructus Lycii 10 ~ 15 parts, Herba Andrographis 10 ~ 30 parts, Radix Achyranthis Bidentatae 5 ~ 20 parts, 10 ~ 30 parts, Poria, Folium Ginkgo 20 ~ 40 parts, 10 ~ 30 parts, Radix Glycyrrhizae, the Radix Astragali 10 ~ 30 parts, Herba Urticae Cannabinae 5 ~ 20 parts, Herba Equiseti Arvinsis 10 ~ 30 parts, Radix Salviae Miltiorrhizae 5 ~ 20 parts, chopping, clean after spend the night by cold water soak, add the purified water of raw material weight 15 times again, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect medicinal liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out precipitate with ethanol, precipitation is rough herbal polysaccharide, and supernatant is rough Chinese medicine flavone;
D, with rough herbal polysaccharide in sterilizing purified water washing c, dissolves with 42 DEG C of sterilizing purified water of 20 times amount after washing, after dissolving, add active carbon 4 DEG C of attach overnight in 0.5% ratio, after absorption through 10000rpm centrifugal 10 minutes, discard precipitation, collection supernatant;
E, by supernatant in d after 0.22um membrane filtration is degerming, carry out 10 times and concentrate, obtained herbal polysaccharide concentrated solution;
F, herbal polysaccharide concentrated solution in e is carried out vacuum lyophilization namely obtain herbal polysaccharide;
G, Chinese medicine flavone rough in c is carried out reduced vacuum concentrate, concentrate time sterilized water for injection dilution cleaning, again carries out reduced vacuum and concentrates, after three times like this, more degerming through 0.22um membrane filtration, then namely obtain Chinese medicine flavone through lyophilisation.
The preparation method of described a kind of infectious bursal disease compound effervescent tablet, is characterized in that taking each raw material of Chinese medicine by Radix Codonopsis 14 ~ 16 parts, Radix Ginseng 8 ~ 12 parts, Fructus Lycii 12 ~ 14 parts, Herba Andrographis 15 ~ 25 parts, Radix Achyranthis Bidentatae 10 ~ 15 parts, 20 ~ 25 parts, Poria, Folium Ginkgo 25 ~ 35 parts, 15 ~ 25 parts, Radix Glycyrrhizae, the Radix Astragali 15 ~ 25 parts, Herba Urticae Cannabinae 10 ~ 15 parts, Herba Equiseti Arvinsis 20 ~ 25 parts, Radix Salviae Miltiorrhizae 8 ~ 12 parts in described step a.
The present invention has following beneficial effect:
1, the infectious bursal disease combination vaccine effervescent tablet prepared by the present invention, reduces the movement requirement of product, adds the convenience that user uses.
2, the infectious bursal disease combination vaccine effervescent tablet prepared by the present invention can carry out feeding of microbial ecological agent while immune infections chicken cloacal bursa live vaccine, decreases manpower and materials, decrease to chicken group stress.
3, the infectious bursal disease combination vaccine effervescent tablet prepared by the present invention can strengthen immunity for chickens power, improves the overall immune effect of chicken group, reduces chicken group antibody dispersion, makes chicken group antibody remain on same level, reduce the risk that chicken group catches.
Detailed description of the invention
In order to make the present invention easier to understand, below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention, NM specific experiment method in the following example, experimental technique carries out routinely usually.
Embodiment 1: cultivate infections chicken cloacal bursa virus by DF-1 cell line and prepare infections chicken cloacal bursa virus suspension
(1) the going down to posterity and cultivation of seedling cell: get chick embryo fibroblast continuous cell line DF-1 cell line through pancreatin cell dispersal liquid had digestive transfer culture, continuing to cultivate with cell growth medium, when forming good monolayer, going down to posterity or virus inoculation for continuing;
(2) breeding of cell seed culture of viruses: get production infectious bursa of Fabricius virus B87 strain (purchased from China Veterinery Drug Inspection Office) seed culture of viruses, being inoculated in by 1% of maintenance medium volume grows up in the chick embryo fibroblast passage cell of good monolayer, put 37 ~ 38 DEG C to continue to cultivate in maintenance medium, harvesting liquid when cytopathy variability reaches more than 75%;
(3) breeding of seedling venom: get in step (1) the above-mentioned cell line culture bottle having formed good monolayer, discard cell growth medium, the maintenance medium containing the cell adapted malicious B87 strain of 1% fabricius bursa that inoculation step (2) obtains, put 37 ~ 38 DEG C to continue to cultivate, venom is gathered in the crops when cytopathy reaches more than 75%, less than-15 DEG C preservations, obtain infections chicken cloacal bursa virus suspension.
Described cell growth medium is containing 90%DMEM/F12,10% hyclone, 90 ~ 110U/ml antibiotics, pH 7.2 ~ 7.3, DO 30 ~ 60%, cell maintenance medium contains 98%DMEM/F12,2% hyclone, 90 ~ 100U/ml antibiotics, pH 7.2 ~ 7.3, DO 30 ~ 50%.
Embodiment 2: the preparation of probiotic bacteria mixed bacteria liquid
(1) cultivation of strain
The cultivation of lactobacillus
MRS fluid medium is preheated to 37 DEG C, simultaneously by 2% inoculum concentration inoculating lactic acid bacterium seed liquor, 37 DEG C of quiescent culture 18 ~ 24 hours, results bacterium liquid saves backup to-4 DEG C;
The cultivation of bacillus cereus
Bacillus culture medium is preheated to 37 DEG C, simultaneously by 1% inoculum concentration inoculation bacillus cereus seed liquor, 37 DEG C of quiescent culture 18 ~ 24 hours, results bacterium liquid to 4 DEG C saves backup;
Described lactobacillus bacterium is: bifidobacterium bifidum (
bifidobacterium bifidum), lactobacillus lactis (
lactobacillus lactis), Lactobacillus buchneri (
lactobacillus buchneri), bifidobacterium longum (
bifidobacterium longum) in one or more;
Described bacillus cereus is: Bacillus licheniformis (
bacillus licheniformis), Bacillus coagulans (
bacillus coagulans), bacillus subtilis (
bacillus subtilis) in one or more;
Described lactobacillus MRS culture medium is:
Peptone 10.0 g, Carnis Bovis seu Bubali cream 10.0 g, yeast extract 5.0 g, diammonium hydrogen citrate 2.0 g, glucose 20.0 g, Tween 80 1.0ml, sodium acetate 5.0 g, dipotassium hydrogen phosphate 2.0 g, magnesium sulfate 0.58 g, manganese sulfate 0.25 g, agar 18.0 g, distilled water 1000 ml, pH6.2-6.6;
Described bacillus culture medium is:
Glucose 20g, peptone 15g, Carnis Bovis seu Bubali cream 0.5g, sodium chloride 5g, MnSo
41g, dipotassium hydrogen phosphate 3g, potassium dihydrogen phosphate 1.5g, magnesium sulfate 0.5g, iron chloride 0.1g, calcium carbonate 0.1g, distilled water 1000ml, 116 DEG C, autoclaving 25 minutes;
(2) mixing of bacterium liquid
To the lactic acid bacterial liquid and the mixing of bacillus cereus bacterium liquid that obtain be cultivated, and adjust bacterium number, obtain probiotic bacteria mixed bacteria liquid.
Embodiment 3: the preparation of herbal polysaccharide and Chinese medicine flavone
A, take each raw material of Chinese medicine by Radix Codonopsis 15 parts, Radix Ginseng 18 parts, Fructus Lycii 13 parts, Herba Andrographis 20 parts, Radix Achyranthis Bidentatae 15 parts, 20 parts, Poria, Folium Ginkgo 30 parts, 20 parts, Radix Glycyrrhizae, the Radix Astragali 20 parts, Herba Urticae Cannabinae 15 parts, Herba Equiseti Arvinsis 20 parts, Radix Salviae Miltiorrhizae 15 parts, chopping, clean after spend the night by cold water soak, add the purified water of raw material weight 15 times again, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect medicinal liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out precipitate with ethanol, precipitation is rough herbal polysaccharide, and supernatant is rough Chinese medicine flavone;
D, with rough herbal polysaccharide in sterilizing purified water washing c, dissolves with 42 DEG C of sterilizing purified water of 20 times amount after washing, after dissolving, add active carbon 4 DEG C of attach overnight in 0.5% ratio, after absorption through 10000rpm centrifugal 10 minutes, discard precipitation, collection supernatant;
E, by supernatant in d after 0.22um membrane filtration is degerming, carry out 10 times and concentrate, obtained herbal polysaccharide concentrated solution;
F, herbal polysaccharide concentrated solution in e is carried out vacuum lyophilization namely obtain herbal polysaccharide;
G, Chinese medicine flavone rough in c is carried out reduced vacuum concentrate, concentrate time sterilized water for injection dilution cleaning, again carries out reduced vacuum and concentrates, after three times like this, more degerming through 0.22um membrane filtration, then namely obtain Chinese medicine flavone through lyophilisation.
Also each raw material of Chinese medicine can be taken according to Radix Codonopsis 10 parts, Radix Ginseng 5 parts, Fructus Lycii 10 parts, Herba Andrographis 10 parts, Radix Achyranthis Bidentatae 20 parts, 30 parts, Poria, Folium Ginkgo 20 parts, 10 parts, Radix Glycyrrhizae, the Radix Astragali 30 parts, Herba Urticae Cannabinae 5 parts, Herba Equiseti Arvinsis 30 parts, Radix Salviae Miltiorrhizae 5 parts in this embodiment; Or take each raw material of Chinese medicine according to Radix Codonopsis 20 parts, Radix Ginseng 20 parts, Fructus Lycii 15 parts, Herba Andrographis 30 parts, Radix Achyranthis Bidentatae 5 parts, 10 parts, Poria, Folium Ginkgo 40 parts, 30 parts, Radix Glycyrrhizae, the Radix Astragali 10 parts, Herba Urticae Cannabinae 20 parts, Herba Equiseti Arvinsis 10 parts, Radix Salviae Miltiorrhizae 20 parts.
Embodiment 4: the preparation of freeze drying protectant
Freeze drying protectant is comprise following component in every 1000ml phosphate buffer: herbal polysaccharide 40g, Chinese medicine flavone 30g, vitamin combination 8g(vitamin C 1.6g, vitamin E 0.8g, vitamin D3 0.6g, vitamin B1 2.2g, vitamin K 1.6g, vitamin B2 1.2g), gelatin 5g, sucrose 50g; Wherein herbal polysaccharide and Chinese medicine flavone are obtained by embodiment 3.
Preparation method: accurately take each component, is dissolved in the phosphate buffer solution of 1000ml, degerming through 0.22um membrane filtration under aseptic condition, and 4 DEG C save backup.
Phosphate buffer by sodium chloride 6 ~ 10g, potassium chloride 0.05 ~ 0.5g, sodium hydrogen phosphate 1 ~ 1.2g, potassium dihydrogen phosphate 0.05 ~ 0.5g, calcium chloride 0.05 ~ 0.2g, the magnesium chloride 0.05 ~ 0.2g containing 6 water of crystallization is dissolved in 1000ml distilled water, through 116 DEG C, autoclaving obtains for 30 minutes.
In this embodiment, freeze drying protectant also can adopt herbal polysaccharide 30g, Chinese medicine flavone 50g, vitamin to combine 6g(vitamin C 1.2g, vitamin E 1.8g, vitamin D3 0.9g, vitamin B1 1.2g, vitamin K 0.6g, vitamin B2 0.3g), gelatin 4g and sucrose 70g; Or polysaccharide 80g, Chinese medicine flavone 20g, vitamin combination 10g(vitamin C 3.5g, vitamin E 1g, vitamin D3 0.5g, vitamin B1 1.5g, vitamin K 2g, vitamin B2 1.5g), gelatin 6g and sucrose 40g.
Embodiment 5: the preparation of effervescent materials
The preparation of a, acid material
Take citric acid 15 parts, lactose 10 parts, EDTA 15 parts, light blue indicator 0.5 part respectively by mass parts, fully mixed by mentioned component, cross 8 ~ 12 mesh sieves after 40 ~ 65 DEG C of oven dry, obtained acid particles is for subsequent use;
The preparation of b, basic matterial
Take sodium bicarbonate 15 parts, lactose 10 parts, light blue indicator 1 part respectively by mass parts, fully mixed by mentioned component, cross 8 ~ 12 mesh sieves after 40 ~ 65 DEG C of oven dry, obtained alkali grain is for subsequent use;
C, above-mentioned basic matterial to be mixed homogeneously by 1:1 with acid material, and add the levamisole of 0.5 ~ 1 mass parts, obtain effervescent materials through Co-60 radiation sterilization.
Embodiment 6: the preparation of infectious bursal disease combination vaccine effervescent tablet
A, the infections chicken cloacal bursa virus suspension sterilizing PBS that embodiment 1 is obtained is adjusted to viral level is 10
8.5tCID
50/ 1ml;
B, the bacterium number in probiotic bacteria mixed bacteria liquid obtained for embodiment 2 is adjusted to 16,000,000,000/ml(wherein lactobacillus content be 8,000,000,000/ml, bacillus cereus content is 8,000,000,000/ml);
C, by infections chicken cloacal bursa virus suspension in a and the probiotic bacteria mixed bacteria liquid in b by volume 1:1 mix;
D, by mixed complex by volume 1:1 add the obtained freeze drying protectant of embodiment 4, through lyophilisation after subpackage, obtain infectious bursal disease compound lyophilized powder;
Prepared by e, effervescent tablet
After the effervescent materials obtained with embodiment 5 by the infectious bursal disease compound lyophilized powder of above-mentioned preparation being mixed homogeneously by weight 2:1 ~ 3, be pressed into tablet with tablet machine, quantitative separating.
The application of test example 1 infectious bursal disease compound effervescent tablet of the present invention
1 material
1.1 infectious bursal disease compound effervescent tablet
Obtain by the embodiment of the present invention 6.
1.2 infectious bursal disease live-vaccine effervescent tablets
Obtained by infections chicken cloacal bursa virus lyophilized formulations and infectious bursal disease live-vaccine effervescent tablet immunological adjuvant.
1.3 tests chicken 200 age in days sea blue laying breed 4000; 10 age in days Sanhuang broilers 4000.
2 methods
4000 200 blue laying breed in age in days sea are divided into four groups, first group 1000 by 2.1, as experiment one group, and infectious bursal disease compound effervescent tablet of the present invention 5 of feeding while complete feed of feeding (with reference to fowl raising standard NY/33-2004); Second group 1000, as experiment two groups, infectious bursal disease compound effervescent tablet of the present invention 10 of feeding while complete feed of feeding (with reference to fowl raising standard NY/33-2004); 3rd group 1000, one group in contrast, feed not containing the infectious bursal disease live-vaccine effervescent tablet 10 of microbial ecological agent while complete feed of feeding (with reference to fowl raising standard NY/33-2004).4th group 1000, two groups in contrast, complete feed of only feeding.The indexs such as laying rate, rate of fertilization, death rate, infections chicken cloacal bursa antibody horizontal, newcastle epidemic disease antibody level respectively organized in test implementation period record, and the observation period is surrounding.
4000 10 age in days Sanhuang broilers are divided into four groups, first group 1000 by 2.2, as experiment one group, and infectious bursal disease compound effervescent tablet of the present invention 5 of feeding while complete feed of feeding (with reference to fowl raising standard NY/33-2004); Second group 1000, as experiment two groups, infectious bursal disease compound effervescent tablet of the present invention 10 of feeding while complete feed of feeding (with reference to fowl raising standard NY/33-2004); 3rd group 1000, one group in contrast, feed not containing the infectious bursal disease live-vaccine effervescent tablet 10 of microbial ecological agent while complete feed of feeding (with reference to fowl raising standard NY/33-2004).4th group 1000, two groups in contrast, complete feed of only feeding.Average daily gain, death rate and infections chicken cloacal bursa antibody horizontal, antibody IgY against chicken Newcastle Disease level respectively organized in test implementation period record, and the observation period is 4 weeks.
3 results
3.1 disintegration result
Get 1 effervescent tablet, put and fill in the 250ml beaker of 200ml water, water temperature 15-25 DEG C, observe and record the consoluet time, the results are shown in Table 1.
Table 1 assay disintegration
The 3.2 200 blue laying breed result of the tests in age in days sea, in table 2.
The blue laying breed result of the test in table 2 200 age in days sea
3.3 10 age in days Sanhuang broiler result of the tests, in table 3.
Table 3 10 age in days Sanhuang broiler result of the test
4 conclusions
In sum, infectious bursal disease compound effervescent tablet of the present invention carries out feeding of microbial ecological agent while immune infections chicken cloacal bursa live vaccine, can improve immunity of organisms.Infections chicken cloacal bursa antibody titer can be improved when chicken group uses, improve laying rate, improve fertility rate of hatching egg, improve broiler daily gain, reduce chicken group death rate.The use of infectious bursal disease compound effervescent tablet of the present invention improves the overall immune effect of chicken group, reduces chicken group antibody dispersion, makes chicken group antibody remain on same level, reduce the risk that chicken group catches.
Claims (6)
1. a preparation method for infectious bursal disease compound effervescent tablet, is characterized in that comprising following processing step:
1) the going down to posterity and cultivation of seedling cell: get chick embryo fibroblast continuous cell line DF-1 cell line through pancreatin cell dispersal liquid had digestive transfer culture, continuing to cultivate with cell growth medium, when forming good monolayer, going down to posterity or virus inoculation for continuing in culture bottle;
2) breeding of cell seed culture of viruses: get production infectious bursa of Fabricius virus B87 strain, being inoculated in by 1% of maintenance medium volume grows up in the chick embryo fibroblast passage cell of good monolayer, put 37 ~ 38 DEG C to continue to cultivate in maintenance medium, harvesting liquid when cytopathy variability reaches more than 75%;
3) breeding of seedling venom: get the cell line culture bottle having formed good monolayer in step 1), discard cell growth medium, inoculation step 2) obtain containing the maintenance medium of the cell adapted malicious B87 strain of 1% fabricius bursa, put 37 ~ 38 DEG C to continue to cultivate, venom is gathered in the crops when cytopathy reaches more than 75%, less than-15 DEG C preservations, obtain infections chicken cloacal bursa virus suspension;
4) preparation of probiotic bacteria mixed bacteria liquid: extracting lactic acid bacterium and bacillus cereus are cultivated respectively, then will the lactic acid bacterial liquid and the mixing of bacillus cereus bacterium liquid that obtain be cultivated, and adjust bacterium number, obtain probiotic bacteria mixed bacteria liquid;
5) lyophilizing of mix preparation: the probiotic bacteria mixed bacteria liquid that the infections chicken cloacal bursa virus suspension and the step 4) that step 3) are obtained obtain aseptically by volume 1:1 mix homogeneously, 1:1 adds freeze drying protectant mix homogeneously by volume again, through lyophilisation after subpackage, obtain mix preparation lyophilized powder, described freeze drying protectant is containing herbal polysaccharide 30 ~ 80g in every 1000ml phosphate buffer, Chinese medicine flavone 20 ~ 50g, vitamin combination 6 ~ 10g, gelatin 4 ~ 6g and sucrose 40 ~ 70g, described herbal polysaccharide and Chinese medicine flavone extract and obtain from following Chinese medicine composition: Radix Codonopsis 10 ~ 20 parts, Radix Ginseng 5 ~ 20 parts, Fructus Lycii 10 ~ 15 parts, Herba Andrographis 10 ~ 30 parts, Radix Achyranthis Bidentatae 5 ~ 20 parts, 10 ~ 30 parts, Poria, Folium Ginkgo 20 ~ 40 parts, 10 ~ 30 parts, Radix Glycyrrhizae, the Radix Astragali 10 ~ 30 parts, Herba Urticae Cannabinae 5 ~ 20 parts, Herba Equiseti Arvinsis 10 ~ 30 parts, Radix Salviae Miltiorrhizae 5 ~ 20 parts, described vitamin combination is containing vitamin C 20 ~ 40%, vitamin E 10 ~ 30%, vitamin D3 5 ~ 15%, vitamin B1 15 ~ 35%, vitamin K 10 ~ 20%, vitamin B2 5 ~ 20%,
6) effervescent tablet preparation: mix preparation lyophilized powder step 5) obtained is mixed homogeneously with effervescent materials, tabletting, and subpackage obtains infectious bursal disease compound effervescent tablet.
2. the preparation method of a kind of infectious bursal disease compound effervescent tablet as claimed in claim 1, to it is characterized in that in described step 4) lactobacillus be bifidobacterium bifidum (
bifidobacterium bifidum), lactobacillus lactis (
lactobacillus lactis), Lactobacillus buchneri (
lactobacillus buchneri), bifidobacterium longum (
bifidobacterium longum) in one or more, described bacillus cereus be Bacillus licheniformis (
bacillus licheniformis), Bacillus coagulans (
bacillus coagulans), bacillus subtilis (
bacillus subtilis) in one or more.
3. the preparation method of a kind of infectious bursal disease compound effervescent tablet as claimed in claim 1, it is characterized in that in described step 4), lactobacillus and bacillus cereus are cultivated according to following steps respectively: MRS fluid medium is preheated to 37 DEG C, simultaneously by 2% inoculum concentration inoculating lactic acid bacterium seed liquor, 37 DEG C of quiescent culture 18 ~ 24 hours, results bacterium liquid to 4 DEG C saves backup; Bacillus culture medium is preheated to 37 DEG C, simultaneously by 1% inoculum concentration inoculation bacillus cereus seed liquor, 37 DEG C of quiescent culture 18 ~ 24 hours, results bacterium liquid to 4 DEG C saves backup.
4. the preparation method of a kind of infectious bursal disease compound effervescent tablet as claimed in claim 1, is characterized in that described freeze drying protectant is containing herbal polysaccharide 50 ~ 70g, Chinese medicine flavone 30 ~ 40g, vitamin combination 7 ~ 9g, gelatin 4.5 ~ 5.5g and sucrose 50 ~ 60g in every 1000ml phosphate buffer.
5. the preparation method of a kind of infectious bursal disease compound effervescent tablet as claimed in claim 1, is characterized in that described herbal polysaccharide and Chinese medicine flavone are obtained by following steps:
A, take each raw material of Chinese medicine by Radix Codonopsis 10 ~ 20 parts, Radix Ginseng 5 ~ 20 parts, Fructus Lycii 10 ~ 15 parts, Herba Andrographis 10 ~ 30 parts, Radix Achyranthis Bidentatae 5 ~ 20 parts, 10 ~ 30 parts, Poria, Folium Ginkgo 20 ~ 40 parts, 10 ~ 30 parts, Radix Glycyrrhizae, the Radix Astragali 10 ~ 30 parts, Herba Urticae Cannabinae 5 ~ 20 parts, Herba Equiseti Arvinsis 10 ~ 30 parts, Radix Salviae Miltiorrhizae 5 ~ 20 parts, chopping, clean after spend the night by cold water soak, add the purified water of raw material weight 15 times again, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect medicinal liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out precipitate with ethanol, precipitation is rough herbal polysaccharide, and supernatant is rough Chinese medicine flavone;
D, with rough herbal polysaccharide in sterilizing purified water washing c, dissolves with 42 DEG C of sterilizing purified water of 20 times amount after washing, after dissolving, add active carbon 4 DEG C of attach overnight in 0.5% ratio, after absorption through 10000rpm centrifugal 10 minutes, discard precipitation, collection supernatant;
E, by supernatant in d after 0.22um membrane filtration is degerming, carry out 10 times and concentrate, obtained herbal polysaccharide concentrated solution;
F, herbal polysaccharide concentrated solution in e is carried out vacuum lyophilization namely obtain herbal polysaccharide;
G, Chinese medicine flavone rough in c is carried out reduced vacuum concentrate, concentrate time sterilized water for injection dilution cleaning, again carries out reduced vacuum and concentrates, after three times like this, more degerming through 0.22um membrane filtration, then namely obtain Chinese medicine flavone through lyophilisation.
6. the preparation method of a kind of infectious bursal disease compound effervescent tablet as claimed in claim 5, is characterized in that taking each raw material of Chinese medicine by Radix Codonopsis 14 ~ 16 parts, Radix Ginseng 8 ~ 12 parts, Fructus Lycii 12 ~ 14 parts, Herba Andrographis 15 ~ 25 parts, Radix Achyranthis Bidentatae 10 ~ 15 parts, 20 ~ 25 parts, Poria, Folium Ginkgo 25 ~ 35 parts, 15 ~ 25 parts, Radix Glycyrrhizae, the Radix Astragali 15 ~ 25 parts, Herba Urticae Cannabinae 10 ~ 15 parts, Herba Equiseti Arvinsis 20 ~ 25 parts, Radix Salviae Miltiorrhizae 8 ~ 12 parts in described step a.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1528453A (en) * | 2001-12-21 | 2004-09-15 | 卫广森 | Heat resisting lyophilized protectant for chicken infections Fabianite bursa hypotpxicity (B87) lyophilized vaccine and preparing process thereof |
| CN101933909A (en) * | 2010-09-15 | 2011-01-05 | 河南省康星药业有限公司 | Vaccine adjuvant effervescent tablet and preparation method thereof |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1528453A (en) * | 2001-12-21 | 2004-09-15 | 卫广森 | Heat resisting lyophilized protectant for chicken infections Fabianite bursa hypotpxicity (B87) lyophilized vaccine and preparing process thereof |
| CN101933909A (en) * | 2010-09-15 | 2011-01-05 | 河南省康星药业有限公司 | Vaccine adjuvant effervescent tablet and preparation method thereof |
Non-Patent Citations (5)
| Title |
|---|
| .2012,第14页第1段-第33页最后1段. * |
| 中国优秀硕士论文全文数据库 农业科技辑> * |
| 中药作为疫苗佐剂的研究概况;罗燕等;《中国兽医杂志》;20091231;第45卷(第8期);61-63 * |
| 复合微生态制剂对肉仔鸡免疫功能的影响;张超范等;《动物医学进展》;20041231;第25卷(第4期);摘要，第102页1.4疫苗、1.5实验方法和1.6疫苗免疫部分、第103页2.3益生菌对肉仔鸡血清HI抗体效价的测定结果部分 * |
| 王永生.应用DF-1细胞和激流式生物反应器大规模繁殖IBDV的研究.< * |
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Denomination of invention: Preparation method of compound effervescent tablets for treating chicken infectious bursal disease Effective date of registration: 20191211 Granted publication date: 20150304 Pledgee: Agricultural Bank of China Limited by Share Ltd. Jinhua Economic Development Zone Branch Pledgor: ZHEJIANG MIBOLERONE BIOLOGICAL TECHNOLOGY Co.,Ltd. Registration number: Y2019330000279 |
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Date of cancellation: 20221020 Granted publication date: 20150304 Pledgee: Agricultural Bank of China Limited by Share Ltd. Jinhua Economic Development Zone Branch Pledgor: ZHEJIANG MIBOLERONE BIOLOGICAL TECHNOLOGY Co.,Ltd. Registration number: Y2019330000279 |