CN101019940A - Organic composite Chinese medicine immunopotentiator/adjuvant and its prepn process - Google Patents
Organic composite Chinese medicine immunopotentiator/adjuvant and its prepn process Download PDFInfo
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Abstract
The present invention proposes one kind of organic composite Chinese medicine immunopotentiator/adjuvant for preventing and treating animal contagious diseases and its preparation process. The Chinese medicine immunopotentiator/adjuvant is prepared with propolis and eight kinds of Chinese medicinal materials, including astragalus root, fortune euonymus stem or leaf, Dangshen, glossy ganoderma, ginseng, etc. and through compounding the extracted liquid of the Chinese medicinal materials and the alcohol leaching liquid of propolis. It can enhance the immunity of available vaccine and strengthen animal's disease resistance.
Description
Technical field
The present invention relates to a kind of pure Chinese medicine composite immune reinforcing agent/adjuvant of preventing and treating poultry infectious disease vaccine and preparation method thereof.
Background technology
The poultry infectious disease is the disease that serious harm herding is produced, it not only can cause the large quantities of death of poultry, livestock and poultry production performance reduces, animal products downgrade and directly influence peasants and herdsmen's income and foreign trade, and also some zoonosis is also threatening human health.Present stage, control for infectious disease, depend on the immunity inoculation of multiple epidemic disease (bacterium) Seedling and the use of medicine morely, but owing to causing body's immunity, a large amount of for a long time Drug abuse and irrational vaccination descends, particularly at a low ebb the or inhibitory state of immunocyte such as lymphocyte, macrophage; New in recent years in addition livestock and poultry pestilence constantly occurs and the pathogen variation and the virulence enhancing of old eqpidemic disease, causes conventional vaccine immunity inoculation preventive effect not good, and dosage strengthens gradually, and does not reach ideal treatment and preventive effect yet.
For immune protective effect and the drug effect that strengthens vaccine, though have panimmunity reinforcing agent and adjuvant to comprise immunological adjuvant (lipopolysaccharide, mycobacterium bovis bcg, mycobacteria segment), inorganic salts immunopotentiator/adjuvant, the synthetics para-immunity reinforcing agent/immunological adjuvant (liposome, levamisole, glucosan sulfuric acid ester) of oils immunological adjuvant, antibacterial or bacterial origin both at home and abroad.Owing to multiple reasons such as some immunopotentiator/adjuvant cost an arm and a leg, majority still is in laboratory stage; Chemicals para-immunity reinforcing agent exists to some extent and reduces the body fertility performance, causes toxic and side effects and drug residue, and drawback such as be detrimental to health; Vaccine adjuvant is used and comparatively extensively also is mainly mineral oil, yet, be prone to stronger stress behind the injection oil seepage, and because of oily adjuvant mineral oil in fluid in vivo because of energy metabolism long-term existence not, often cause local tissue damage and form the scleroma that is difficult to dissipate, these mineral oil that can't absorb in the poultry body cause that easily carcase quality descends, and directly enter human body through food chain, have a strong impact on the healthy of people.Also there is the exploration and the report of some propolis adjuvant inactivated vaccines fowl infectious disease (as avian cholera, newcastle disease) prevention aspect in addition, but the adjuvant type is single, mostly is simple propolis, and object of prevention is chicken, duck, goose.
Summary of the invention
The applicant examines and carries out on the basis of research work preventing and treating facing of fowl diseases, biological preparation such as poultry vaccine, antibody, immunostimulant and Chinese medicine preparation have been carried out a large amount of research, proposed pure Chinese medicine compound immunopotentiator/adjuvant of a kind of efficient, safe, stable control poultry infectious disease and preparation method thereof, this is a purpose of the present invention.The present invention can be to obtain the comparatively ideal immunopotentiator/adjuvant that is suitable for preventing and treating livestock and poultry than low production cost, it can strengthen the immunity of existing vaccine, can strengthen the nonspecific defense mechanism of poultry resistance against diseases again.
Realize that the technical scheme that the object of the invention adopted is as follows:
1, composite immune reinforcing agent/adjuvant prescription screening
Utilize Chinese medicines such as propolis, Caulis Seu Folium Euonymi Fortunei, the Radix Astragali, Radix Codonopsis, Ganoderma, contain compositions such as multiple flavonoid, saponins, amino acids, alkaloid, phenolic compound, phyllins, carotene, extract active ingredient by certain technology, reasonable combination, as immunological adjuvant good immunological enhancement is arranged, function with QI invigorating, resisting fatigue and raising body reticuloendothelial system, the leukocyte increasing phagocytic function, inducement interferon, the formation of enhancing antibody, improve the specificity and the non-specific immunity of body, the enhancing body premunition.
By Embryo Gallus domesticus neutralization test, animal (chicken) immunity test, from the extracting solution of various Chinese medicines, filter out based on propolis, in conjunction with the Radix Astragali, Caulis Seu Folium Euonymi Fortunei, Radix Codonopsis, Ganoderma, become new prescription with the certain proportion organic assembling.
The crude drug prescription of the Chinese medicine compound immunopotentiator/adjuvant that is obtained is (by weight ratio):
0～3 part of 0.01～0.03 part of propolis, 1～3 part of the Radix Astragali, 1～3 part of Caulis Seu Folium Euonymi Fortunei, 1～3 part of Radix Codonopsis, 1～3 part of Ganoderma, 0～3 part of Radix Ginseng, 0～3 part of Rhizoma Coptidis, 0～3 part of Flos Lonicerae and Fructus Forsythiae.
2, the preparation of composite immune reinforcing agent/adjuvant
By above-mentioned prescription weighting raw materials, follow these steps to prepare composite immune reinforcing agent/adjuvant:
A) with the propolis that takes by weighing, the ethanol with 95% soaks molten, makes the propolis-containing mass concentration and be 20%～50% propolis tincture;
B) with the crude drug Radix Astragali, Caulis Seu Folium Euonymi Fortunei, Radix Codonopsis, Ganoderma, Radix Ginseng, Rhizoma Coptidis, Flos Lonicerae and the Fructus Forsythiae that take by weighing, decoct with water respectively 2 times, each 2～3 hours, collecting decoction filters, and filtrate decompression is concentrated, handle 2 times through ethanol precipitation again, after ethanol extract reclaimed ethanol, 4～30 times of amounts of pressing gains weight added injection waters, put 4 ℃ 48 hours, filter, get supernatant, boiled 10 minutes, refilter, get supernatant, regulate pH value to 4.5～5.0 with 10% sodium hydroxide solution, adding tween 80 to its mass concentration is 1%～6%, makes extracting solution;
C) add benzyl alcohol in the gained extracting solution, the mass concentration that makes benzyl alcohol is 1%～5%, mixes with the above-mentioned propolis tincture that makes again, promptly makes organic Chinese medicine composite immune reinforcing agent of the present invention/adjuvant suspension.
At above-mentioned steps b) in, when the organic Chinese medicine complex immunity adjuvant of preparation, after the alcohol extract of described crude drug reclaims ethanol, can add injection waters by 4～10 times of amounts of gains weight; When the organic Chinese medicine composite immune reinforcing agent of preparation, after the alcohol extract of described crude drug reclaims ethanol, can add injection waters by 10～30 times of amounts of gains weight.
This Chinese medicine composite immune reinforcing agent/adjuvant uses as the Chinese medicine composite immune reinforcing agent, can directly give fowl, raise orally, also can make muscle or subcutaneous injection, and using method is as follows:
1. muscle or subcutaneous injection: fowl: 0.5 milliliter～1 milliliter/plumage part (0.5 kilogram weight); Pig: 2 milliliters～4 milliliters/(50 kilogram weights).
2. oral: fowl: 0.5 milliliter～1 milliliter/plumage part (0.5 kilogram weight); Pig: 4 milliliters～8 milliliters/(50 kilogram weights).
The scope of application: birds and pigs such as chicken, duck, goose, Columba livia.
Be mixed as adjuvant and PA/PMV-I (Columba livia fowl I type paramyxovirus) or NDV inactivation antigens such as (viruses of newcastle disease) with this organic Chinese medicine composite immune reinforcing agent/adjuvant and can be mixed with Chinese medicine composite adjuvant inactivated vaccine.Its compound method is as follows:
To transfer to inactivation antigens such as qualified PA/PMV-I or NDV and this Chinese medicine complex immunity adjuvant suspension through titration by 1～3: 1～5 mixed, after the tween 80 of will sterilizing add in the mixed liquor by 1%～2%, mix, impouring homogenate pump emulsifying again, packing after the aseptic and safety inspection, promptly become pure Chinese medicine composite adjuvant inactivated vaccine, be applicable to that muscle or subcutaneous injection use.The using method of Chinese medicine composite adjuvant PA/PMV-I (NDV) inactivated vaccine is as follows:
1. prevention: birdss such as Columba livia, chicken, muscle or 0.5 milliliter/plumage of subcutaneous injection part;
2. urgent epidemic prevention inoculation: Columba livia, chicken, 0.8 milliliter～1.0 milliliters/plumage of muscle or subcutaneous injection part.
The technique effect that the present invention reaches
1. animal experiment 1: getting 60 of the yellow broiler of the healthy Guangdong of 1 age in days fowl is test chicken, is divided into 2 groups (I is a test group, and II is the blank group) 30 every group at random.Test group is when 4～8 ages in days, in drinking-water, press this Chinese medicine of 0.5 milliliter of interpolation of every chicken every day composite immune reinforcing agent, matched group does not then add this preparation and makes blank, and when 14 ages in days, 21 ages in days, 35 ages in days every group get 10 chickens respectively sterile blood sampling sample of back of weighing at random, carry out in lymphocyte transformation test and the blood Superoxide Dismutase and measure; With test chicken jugular vein sacrificed by exsanguination, dissect and separation fabricius bursa and spleen then, weigh respectively, calculate fabricius bursa coefficient and spleen coefficient.The result shows, the thymus index of test group chickling, fabricius bursa index, index and spleen index be all apparently higher than the blank group, between any two inequality heteropole significantly (p＜0.01); Record test group SOD vigor simultaneously when 14 ages in days and matched group significant difference (p＜0.05), test group SOD vigor is compared difference extremely significantly (p＜0.01) with matched group when 21 ages in days, 35 ages in days.The result illustrates that comprehensively this Chinese medicine composite immune reinforcing agent can obviously promote the propagation of immunologically competent cell in the immune organ, improves the function of body cell immunity and humoral immunization.
2. in order to verify this Chinese medicine composite immune reinforcing agent using value in Production of Livestock and Poultry, we add the Chinese medicine composite immune reinforcing agent in feedstuff after, survival rate, gain in weight and the feedstuff-meat ratio of broiler, squab carried out determination test, obtained effect preferably.
2.1. getting 500 of the yellow high quality meat chickens of the healthy Guangdong of 1 age in days fowl is test chicken, is divided into 2 groups at random, 250 every group.Test group is when 4～8 ages in days, and by this Chinese medicine of 0.5 milliliter of interpolation of every chicken every day composite immune reinforcing agent, matched group does not then add this preparation and makes blank in drinking-water, the conventional raising, and experimental period is 30 days.When off-test, weigh, and calculate its survival rate, gain in weight and feedstuff-meat ratio.We find on the survival rate of on average brooding, two indexs of weightening finish, added composite immune reinforcing agent test group (98.6%, 450.65 gram), all greater than the matched group (92.3% that does not add composite immune reinforcing agent, 389.5 gram), through data statistic analysis, difference extremely significantly (P＜0.01) between any two; Feedstuff-meat ratio test group (1.56: 1) is obviously low than matched group (1.72: 1).
2.2. get 50 of 15～16 age in days squabs, be divided at random test group and matched group each 25, from nest to fattening the storehouse feeding for fattening.Simultaneously, establish a repetition for every group.The test group squab is respectively at 20～24 ages in days, and each by only gavaging 0.5 milliliter of composite immune reinforcing agent, every morning, afternoon, each was irritated once.Each organizes squab when 15 ages in days and 35 ages in days, counts respectively and weighs.Result of the test shows that on weightening finish and two indexs of material anharmonic ratio, test group (123.2 grams, 4.86: 1) all is better than matched group (95.2 grams, 6.16: 1).
3. the laboratory potency test of Columba livia paramyxovirus disease (PA/PMV-I) Chinese medicine composite adjuvant inactivated vaccine
Get 240 of the non-immune Columba livias of 3 ages in days, be divided into 2 test group at random, the 1st group of (120) intramuscular inoculation Chinese medicine adjuvant inactivated vaccine 0.5ml/ only, the 2nd group (120) do not carry out vaccine immunity with in contrast.Regular before and after immunity (preceding 1 day of immunity and immunity back 3 days, 6 days, 9 days, 14 days, 21 days, 35 days, 60 days, 90 days, 120 days, 150 days, 180 days) blood drawing 1ml separation of serum is with Columba livia A/PMV-I GDP
199902Be antigen, micromethod is measured its HI valency respectively routinely, and its HI valency result judges that the high dilution to suppress blood clotting fully is a terminal point, with Xlog
2Expression, each mensuration all repeats once, averages.And in immunity back 21 days, 35 days, 90 days, 120 days, 150 days, 180 days, 2 groups were respectively randomly drawed 20, carried out strong poison (200 ELD
50/ 0.1ml) to attack, isolated rearing was observed 25 days continuously, to detect its immune protective effect.Result of the test shows, uses behind the Chinese medicine composite adjuvant can to detect antibody in 4 days, keeps summit (10.8～11.5log in 14 days to 30 days
2), still can keep higher antibody horizontal (6.0～6.9log in 180 days
2), and do not have oil seepage be difficult to inject and the local problem that forms scleroma.In laboratory, test group was used 200 ELD of virulent strain in back 21 days, 60 days, 90 days, 120 days, 150 days, 180 days respectively in immunity
50/ 0.1ml attacks, and its protection index is respectively 100%, 100%, 100%, 94.4%, 88.9%, 83.5%.At this moment, detect its serum antibody HI valency, average (10.5log
2～11.5log
2) log
2More than.According to " test report of HI antibody titer and counteracting toxic substances protective rate parallel relation " we formulate for immunity measure after 21 days HI tire on average should 〉=6log
2
4. the laboratory potency test of newcastle disease (NDV) Chinese medicine composite adjuvant inactivated vaccine
Get 240 of the non-immune chickens of 10 ages in days, be divided into 2 test group at random, the 1st group of (180) intramuscular inoculation Chinese medicine adjuvant inactivated vaccine 0.5ml/ only only therefrom takes out 60 intramuscular inoculation Chinese medicine adjuvant inactivated vaccine 0.8ml/ at random after 21 days; The 2nd group (180) do not carry out vaccine immunity with in contrast.Regular before and after immunity (preceding 1 day of immunity and immunity back 4 days, 6 days, 9 days, 14 days, 21 days, 35 days, 60 days, 90 days, 120 days, 150 days, 180 days) blood drawing 1ml separation of serum, with NDIV is antigen, micromethod is measured its HI valency respectively routinely, its HI valency result judges that the high dilution to suppress blood clotting fully is a terminal point, with Xlog
2Expression, each mensuration all repeats once, averages.And in immunity back 21 days, 35 days, 90 days, 120 days, 150 days, 180 days, 2 groups were respectively randomly drawed 20, carried out strong poison (200 ELD
50/ 0.1ml) to attack, isolated rearing was observed 25 days continuously, to detect its immune protective effect.Result of the test shows, uses behind this Chinese medicine composite adjuvant can to detect antibody in 4 days, keeps summit (8.8log in 14 days to 35 days
2～10.1log
2), still can keep higher antibody horizontal (6.0log in 6 months
2), and do not have oil seepage be difficult to inject and the local problem that forms scleroma.In laboratory, test group was used 200 ELD of virulent strain in back 21 days, 60 days, 90 days, 120 days, 150 days, 180 days respectively in immunity
50/ 0.1ml attacks, and its protection index is respectively 100%, 100%, 100%, 90%, 85.4%, 81.9%; 200 ELD of virulent strain were used in two immunity in back 120 days, 150 days, 180 days respectively
50/ 0.1ml attacks, and its protection index is respectively 100%, 93%, 85.4%.
As seen, organic Chinese medicine composite immune reinforcing agent/adjuvant of the present invention has following effect and characteristics as the Chinese medicine composite immune reinforcing agent to poultry:
1. make its immunity that can strengthen vaccine, can strengthen the nonspecific defense mechanism of poultry resistance against diseases again;
2. disease-prevention health effect: double recuperation, preventable disease takes place, and the order of severity that palliates a disease and take place improves survival rate 8.9%～13.8%;
3. growth promoting function: can make the fowl poultry look faster, bigger;
4. anti-stress effect: can significantly reduce fowl poultry stress;
5. reduce the diseases prevention and treatment cost: can reduce diseases prevention and treatment cost 50%.
6. widely applicable: as can be widely used in precious special fowl such as domestic birds and animals and Columba livia, Gallus jabouillei;
7. remarkable in economical benefits:, increase economic efficiency because above-mentioned many-sided effect can reduce the comprehensive feeding cost significantly.
With this organic Chinese medicine composite immune reinforcing agent/adjuvant as the pure Chinese medicine composite adjuvant inactivated vaccine of adjuvant preparation through field trial, effect is as follows:
1.PA/PMV-I the effect of field application of Chinese medicine composite adjuvant inactivated vaccine
In order to observe the effect of field application of PA/PMV-I Chinese medicine composite adjuvant inactivated vaccine, we have successively produced 10,000 plumage parts respectively, 10,000 Columba livias in 6 fields, different regions are used in Guangdong Province, after the Columba livia that makes preparations for sowing go up in 30～45 ages in days and pairing and respectively inoculate 1 time in 1～2 week of cage, epidemic situation did not take place in half a year; Plant Columba livia field (2 fields) preferably in the raising condition, inoculate and produce 4000 pairs of Columba livias more than 2 ages, in 1 year Columba livia newcastle epidemic situation does not take place, the prevention effective percentage can reach 100%; In the vulnerable area, the squab of 4 batches 3～15 ages in days of inoculation, totally 1000, safe and effective, epidemic situation did not take place in campaign whole meat pigeon, also improve its survival rate and productivity effect; To this patient is taken place, after promptly inoculating, but 4～7 days basic controlling epidemic situations, except being in a bad way individually, outside the sick Columba livia of torticollis occurs, turning round serious nervous symptoms such as neck was difficult to cure, remaining all rehabilitation gradually can reach more than 92.3% the urgent epidemic prevention inoculation effective percentage of primary disease.6 Columba livia field immune effects are observed and the result of sampling Detection antibody horizontal shows that through exempting from this vaccine after half a year, its average HI valency can reach 5.6log through following the tracks of
2More than.
2.NDV the effect of field application of Chinese medicine composite adjuvant inactivated vaccine
In order to observe the effect of field application of NDV Chinese medicine composite adjuvant inactivated vaccine, we have successively produced 5000 plumage parts respectively, 3000 broiler in 3 fields, different regions are used in Guangdong Province, can reach 100% to the prevention effective percentage of newcastle disease, and the effective percentage of promptly preventing epidemic can reach more than 90.1%.
The specific embodiment
Embodiment one
The crude drug prescription of Chinese medicine compound immunopotentiator/adjuvant is (by weight ratio):
1 part of the Radix Astragali, 1 part of Caulis Seu Folium Euonymi Fortunei, 1 part of Radix Codonopsis, 1 part of Ganoderma, 0.01 part of pure propolis.
The preparation of Chinese medicine compound immunopotentiator/adjuvant:
1, pure propolis is soaked molten with 95% ethanol, make the propolis-containing mass concentration and be 98% propolis tincture;
2, with the Radix Astragali, Caulis Seu Folium Euonymi Fortunei, Radix Codonopsis and Ganoderma, boil 2 times through decocting respectively, each 2～3 hours, collecting decoction filtered, filtrate decompression is concentrated, handle 2 times through ethanol precipitation again, alcohol extract is reclaimed ethanol after, in gains, add injection water (when the preparation immunological adjuvant, add 5 parts of injection waters by every weight portion gains, when preparation during immunostimulant, add 12 parts of injection waters by every weight portion gains), put 4 ℃ 48 hours, filter, after boiled 10 minutes, refilter, regulate pH value to 4.5～5.0 with 10% sodium hydroxide solution, adding tween 80 to its mass concentration is 2%, makes extracting solution;
3, add benzyl alcohol in the gained extracting solution, the mass concentration that makes benzyl alcohol is 3%, mixes with the above-mentioned propolis tincture that makes again, promptly makes organic Chinese medicine composite immune reinforcing agent of the present invention/adjuvant suspension.
The preparation of PA/PMV-I (NDV) Chinese medicine composite adjuvant inactivated vaccine
After transferring to qualified PA/PMV-I (NDV) antigen, titration mixes with Chinese medicine compound adjuvant suspension 1: 1～1.5, after the tween 80 of will sterilizing add in the suspension by 1%～2%, mix, impouring homogenate pump emulsifying again, after the aseptic and safety inspection, packing becomes pure Chinese medicine composite adjuvant PA/PMV-I (NDV) inactivated vaccine.
Quality index: this Chinese medicine compound adjuvant PA/PMV-I (NDV) inactivated vaccine and Chinese medicine compound immunostimulant are the brownish black suspension, good evenness, and viscosity is low, and syringeability is good; Deposit more than February under 4 ℃, the bottle end, have a little precipitation, but shake up before use, not impact effect; Do not contain other cause of disease; Aseptic; Safe in utilization, effectively.
Embodiment two
The crude drug prescription of Chinese medicine compound immunopotentiator/adjuvant is (by weight ratio):
3 parts of the Radixs Astragali, 3 parts of Caulis Seu Folium Euonymi Fortuneis, 3 parts of Radix Ginsengs, 3 parts of Radix Codonopsis, 3 parts of Ganodermas, 3 parts of Rhizoma Coptidis, 3 parts of Flos Loniceraes, 3 parts of Fructus Forsythiaes, 0.03 part of pure propolis.
The preparation of Chinese medicine compound immunopotentiator/adjuvant:
1, pure propolis is soaked molten with 95% ethanol, make the propolis-containing mass concentration and be 98% propolis tincture;
2, with the Radix Astragali, Caulis Seu Folium Euonymi Fortunei, Radix Ginseng, Radix Codonopsis, Ganoderma, Rhizoma Coptidis, Flos Lonicerae and Fructus Forsythiae, boil 2 times through decocting respectively, each 2～3 hours, collecting decoction filtered, filtrate decompression is concentrated, handle 2 times through ethanol precipitation again, alcohol extract is reclaimed ethanol after, in gains, add injection water (when the preparation immunological adjuvant, add 8 parts of injection waters by every weight portion gains, when preparation during immunostimulant, add 20 parts of injection waters by every weight portion gains), put 4 ℃ 48 hours, filter, after boiled 10 minutes, refilter, regulate pH value to 4.5～5.0 with 10% sodium hydroxide solution, adding tween 80 to its mass concentration is 2%, makes extracting solution;
3, add benzyl alcohol in the gained extracting solution, the mass concentration that makes benzyl alcohol is 4%, mixes with the above-mentioned propolis tincture that makes again, promptly makes organic Chinese medicine composite immune reinforcing agent of the present invention/adjuvant suspension.
The preparation of PA/PMV-I (NDV) Chinese medicine composite adjuvant inactivated vaccine
After transferring to qualified PA/PMV-I (NDV) antigen, titration mixes with Chinese medicine compound adjuvant suspension 1: 1～1.5, after the tween 80 of will sterilizing add in the suspension by 1%～2%, mix, impouring homogenate pump emulsifying again, after the aseptic and safety inspection, packing becomes pure Chinese medicine composite adjuvant PA/PMV-I (NDV) inactivated vaccine.
Embodiment three
The crude drug prescription of Chinese medicine compound immunopotentiator/adjuvant is (by weight ratio):
1 part of the Radix Astragali, 1 part of Caulis Seu Folium Euonymi Fortunei, 2 parts of Radix Codonopsis, 2 parts of Ganodermas, 1 part of Rhizoma Coptidis, 1 part of Flos Lonicerae, 0.02 part of pure propolis.
The preparation of Chinese medicine compound immunopotentiator/adjuvant:
1, pure propolis is soaked molten with 95% ethanol, make the propolis-containing mass concentration and be 98% propolis tincture;
2, with the Radix Astragali, Caulis Seu Folium Euonymi Fortunei, Radix Codonopsis, Ganoderma, Rhizoma Coptidis, Flos Lonicerae and Fructus Forsythiae, boil 2 times through decocting respectively, each 2～3 hours, collecting decoction filtered, filtrate decompression is concentrated, handle 2 times through ethanol precipitation again, alcohol extract is reclaimed ethanol after, in gains, add injection water (when the preparation immunological adjuvant, add 6 parts of injection waters by every weight portion gains, when preparation during immunostimulant, add 15 parts of injection waters by every weight portion gains), put 4 ℃ 48 hours, filter, after boiled 10 minutes, refilter, regulate pH value to 4.5～5.0 with 10% sodium hydroxide solution, adding tween 80 to its mass concentration is 2%, makes extracting solution;
3, add benzyl alcohol in the gained extracting solution, the mass concentration that makes benzyl alcohol is 3%, mixes with the above-mentioned propolis tincture that makes again, promptly makes organic Chinese medicine composite immune reinforcing agent of the present invention/adjuvant suspension.
The preparation of PA/PMV-I (NDV) Chinese medicine composite adjuvant inactivated vaccine
After transferring to qualified PA/PMV-I (NDV) antigen, titration mixes with Chinese medicine compound adjuvant suspension 1: 1～1.5, after the tween 80 of will sterilizing add in the suspension by 1%～2%, mix, impouring homogenate pump emulsifying again, after the aseptic and safety inspection, packing becomes pure Chinese medicine composite adjuvant PA/PMV-I (NDV) inactivated vaccine.
Embodiment four
The crude drug prescription of Chinese medicine compound immunopotentiator/adjuvant is (by weight ratio):
1 part of the Radix Astragali, 1 part of Caulis Seu Folium Euonymi Fortunei, 2 parts of Radix Codonopsis, 2 parts of Ganodermas, 1 part of Rhizoma Coptidis, 1 part of Flos Lonicerae, 1 part of Fructus Forsythiae.
The compound method of Chinese medicine compound immunostimulant or adjuvant and Chinese medicine composite adjuvant inactivated vaccine thereof such as embodiment three.
The test effect of above-mentioned four Chinese medicine composite adjuvant inactivated vaccines that embodiment prepared:
Getting 60 of the yellow high quality meat chickens of the non-immune health of 7 ages in days Guangdong fowl is test chicken, is divided into 5 groups at random, 10 every group.Test (I-IV) is organized when 10 ages in days, the pure Chinese medicine composite adjuvant that intramuscular injection is made respectively by following 4 kinds of prescriptions be made into vaccine with-NDV inactivation antigen, the then not immune blank of doing of matched group, the conventional raising, experimental period is 21 days.Regular (preceding 1 day of immunity and immunity back 3 days, 9 days, 14 days, 21 days) blood drawing separation of serum before and after immunity, micromethod is measured its HI valency respectively routinely, and its HI valency result judges that the high dilution to suppress blood clotting fully is a terminal point, with Xlog
2Expression, each mensuration all repeats once, averages.The result is as follows:
Serum N D-HI TPPA result before and after the immunity of Chinese medicine composite adjuvant NDV inactivated vaccine
| Group | Before and after the vaccine immunity (my god) serum N D-HI antibody titer is with nlog 2Expression | ||||
| Immunity preceding 1 | Immunity back 3 | 9 | 14 | 21 | |
| Test I group: embodiment one prescription | 1.9 | 3.0 | 4.9 | 8.8 | 10.5 |
| Test II group: embodiment two prescriptions | 1.0 | 3.0 | 5.3 | 9.0 | 11.0 |
| Test III group: embodiment three prescriptions | 1.5 | 3.3 | 5.0 | 8.3 | 10.0 |
| Test IV group: embodiment four prescriptions | 1.6 | 3.2 | 4.8 | 7.0 | 8.5 |
| The blank group | 1.4 | 1.0 | 0.5 | 0 | 0 |
Claims (5)
1. organic Chinese medicine composite immune reinforcing agent/adjuvant is characterized in that being made by following bulk drugs: 0～3 part of 0.01～0.03 part of propolis, 1～3 part of the Radix Astragali, 1～3 part of Caulis Seu Folium Euonymi Fortunei, 1～3 part of Radix Codonopsis, 1～3 part of Ganoderma, 0～3 part of Radix Ginseng, 0～3 part of Rhizoma Coptidis, 0～3 part of Flos Lonicerae and Fructus Forsythiae.
2. organic Chinese medicine composite immune reinforcing agent/adjuvant according to claim 1 is characterized in that being made by following bulk drugs: 1 part of 0.01 part of propolis, 1 part of the Radix Astragali, 1 part of Caulis Seu Folium Euonymi Fortunei, 1 part of Radix Codonopsis and Ganoderma.
3. the preparation method of the described organic Chinese medicine composite immune reinforcing agent/adjuvant of claim 1 is characterized in that comprising the following steps:
A) take by weighing propolis, the ethanol with 95% soaks molten, makes the propolis-containing mass concentration and be 98% propolis tincture;
B) the weighting raw materials Radix Astragali, Caulis Seu Folium Euonymi Fortunei, Radix Codonopsis, Ganoderma, Radix Ginseng, Rhizoma Coptidis, Flos Lonicerae and Fructus Forsythiae, decoct with water respectively 2 times, each 2～3 hours, collecting decoction filters, and filtrate decompression is concentrated, handle 2 times through ethanol precipitation again, after alcohol extract reclaimed ethanol, 4～30 times of amounts of pressing gains weight added injection waters, put 4 ℃ 48 hours, filter, get supernatant, boiled 10 minutes, refilter, get supernatant, regulate pH value to 4.5～5.0 with 10% sodium hydroxide solution, adding tween 80 to its mass concentration is 1%～6%, makes extracting solution;
C) add benzyl alcohol in the gained extracting solution, the mass concentration that makes benzyl alcohol is 1%～5%, mixes with the above-mentioned propolis tincture that makes again, promptly makes composite immune reinforcing agent/adjuvant suspension.
4. the preparation method of organic Chinese medicine composite immune reinforcing agent/adjuvant according to claim 3, it is characterized in that when the organic Chinese medicine complex immunity adjuvant of preparation, in step b), after the alcohol extract of described crude drug reclaimed ethanol, 4～10 times of amounts of pressing gains weight added injection water.
5. the preparation method of organic Chinese medicine composite immune reinforcing agent/adjuvant according to claim 3, it is characterized in that when the organic Chinese medicine composite immune reinforcing agent of preparation, in step b), after the alcohol extract of described crude drug reclaimed ethanol, 10～30 times of amounts of pressing gains weight added injection water.
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| CNA2007100270636A Pending CN101019940A (en) | 2007-03-06 | 2007-03-06 | Organic composite Chinese medicine immunopotentiator/adjuvant and its prepn process |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102379971A (en) * | 2011-11-22 | 2012-03-21 | 青岛绿曼生物工程有限公司 | Compound propolis composition for treating cage layer fatigue and preparation method thereof |
| CN103520219A (en) * | 2013-10-15 | 2014-01-22 | 胡艳欣 | Nature immunologic adjuvant and vaccine composition thereof, as well as preparation method |
| CN104324153A (en) * | 2014-10-22 | 2015-02-04 | 郑州后羿制药有限公司 | Traditional Chinese medicine composition for improving immunity of animals and preparation method thereof |
| CN104800275A (en) * | 2015-05-04 | 2015-07-29 | 山东正邦生物科技有限公司 | Traditional Chinese medicine polysaccharide immunoenhancer for livestock and poultry |
| CN108148761A (en) * | 2017-12-31 | 2018-06-12 | 天津赫莱恩特生物科技有限公司 | A kind of composition for improving Eimeria Tenella egg capsule yield and preparation method thereof |
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2007
- 2007-03-06 CN CNA2007100270636A patent/CN101019940A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102379971A (en) * | 2011-11-22 | 2012-03-21 | 青岛绿曼生物工程有限公司 | Compound propolis composition for treating cage layer fatigue and preparation method thereof |
| CN102379971B (en) * | 2011-11-22 | 2013-10-23 | 青岛绿曼生物工程有限公司 | Compound propolis composition for treating cage layer fatigue and preparation method thereof |
| CN103520219A (en) * | 2013-10-15 | 2014-01-22 | 胡艳欣 | Nature immunologic adjuvant and vaccine composition thereof, as well as preparation method |
| CN103520219B (en) * | 2013-10-15 | 2019-05-07 | 胡艳欣 | A kind of innate immunity adjuvant and its vaccine composition and preparation method |
| CN104324153A (en) * | 2014-10-22 | 2015-02-04 | 郑州后羿制药有限公司 | Traditional Chinese medicine composition for improving immunity of animals and preparation method thereof |
| CN104800275A (en) * | 2015-05-04 | 2015-07-29 | 山东正邦生物科技有限公司 | Traditional Chinese medicine polysaccharide immunoenhancer for livestock and poultry |
| CN104800275B (en) * | 2015-05-04 | 2019-01-11 | 山东正邦生物科技有限公司 | Livestock and poultry herbal polysaccharide immunopotentiator |
| CN108148761A (en) * | 2017-12-31 | 2018-06-12 | 天津赫莱恩特生物科技有限公司 | A kind of composition for improving Eimeria Tenella egg capsule yield and preparation method thereof |
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