CN108096574A - A kind of porcine pseudorabies virus, pig epidemic diarrhea vaccine combination, preparation method and applications - Google Patents
A kind of porcine pseudorabies virus, pig epidemic diarrhea vaccine combination, preparation method and applications Download PDFInfo
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
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Abstract
The present invention relates to a kind of porcine pseudorabies virus (PRV), Porcine epidemic diarrhea virus (PEDV) vaccine combinations, the vaccine combination includes the porcine pseudorabies virus inactivated vaccine of 24 part immunizing doses and attenuated live vaccines and 12 doses Porcine epidemic diarrhea virus inactivated vaccines.The present invention also provides the vaccine combinations to treat the preparation method and applications of pig epidemic diarrhea disease, the vaccine combination can effectively induce the innate immunity, there is remarkable result in terms of pig epidemic diarrhea disease is treated, and treatment cost is low, and program is simple.
Description
Technical field
The invention belongs to animal virology fields, are related to a kind of porcine pseudorabies virus, Porcine epidemic diarrhea virus vaccine group
Close object and its application in treatment pig epidemic diarrhea.
Background technology
Pig epidemic diarrhea (Porcine Epidemic Diarrhea, PED), is the important intestines for currently endangering pig breeding industry
Tract disease is the contact enteric infectious disease as caused by Porcine epidemic diarrhea virus (PEDV).Sick pig is major source of infection.It is various
The pig at age can infection morbidity, main clinical symptoms for watery diarrhea or diarrhea simultaneously with have vomiting, dehydration.
The weight of symptom is variant with the size at age, and the age is smaller, and symptom is heavier.The incidence of suckling pig is very high, in a week old
Newborn piglet occur diarrhea after 3-4 days, presentation serious dehydration and it is dead, up to 50%, the highest death rate reaches the death rate
100%, retarded growth after a small number of pigs recover.Sow incidence changes very big, about 15-90%, diarrhea about one week.Fertilizer
It educates pig and diarrhea, death rate 1%-3% all occurs after same circle raising infection.
Porcine pseudorabies virus belongs to herpesviral, and there is very strong stimulation body to generate immunization of cell.In disease-resistant original
Aspect, nospecific immunity are bases, its main feature is that immunological effect occurs soon, sphere of action is wide, has very to humoral immunity
Important booster action.The vaccine adjuvant used at present mainly stimulates the stronger innate immunity of body generation or cell to exempt from
Epidemic disease has the function that assisted vaccine.
Many Chinese medicines contain relatively rich sugar, alkaloid, glycoside substance etc., these substances are to nospecific immunity and specifically
Property be immunized and have facilitation, mainly swallowed by transferring the antiviral active factor of non-specific antimicrobial in animal body such as cell,
Bacteriolysin, bacteriocidin (enzyme) etc. enhance immune function.
Research shows that Lycium barbarum polysaccharides can substantially increase mouse peripheral blood T lymphocyte percentages, and is situated between to T lymphocytes
The immune response led has apparent facilitation；Caulis Spatholobi decocting liquid has facilitation to normal splenocytes IL-2's；Radix Codonopsis
The ability of Turnover of Mouse Peritoneal Macrophages phagocytosis chicken red blood cell can be enhanced.
The measure of control PED is mainly vaccine inoculation prevention at present, since popular street strain has differences with vaccine strain, with
And PEDV is separately cultured difficulty, currently on the market without preferable this disease of vaccine prevention.But for the treatment side of morbid pig
Method is seldom, and effect is bad.This disease is invalid using antibiosis extract for treating, and general way is stringent isolation sick pig, is used simultaneously
Drug carries out auxiliary treatment, suggested design：High fever acupuncture needle+canavalin, treatment cost are very high.It is generally recognised that symptom management
Best mode is the pig for slaughtering vomiting and watery diarrhea, this is turned off the infection sources, the way of the susceptible swinery of protection.
At present, domestic-developed Chinese medicine preparation is for treating PED, ' a kind of such as Application No. CN201410791605.7
Treat the Chinese medicine composition of pig epidemic diarrhea ', Application No. CN201210596621.1's is ' a kind of to treat pig epidemic diarrhea
Chinese medicine '.But these treatment prescriptions are related to that medicinal material is more, and cost is higher, and curative effect is general.
The content of the invention
In view of above-mentioned deficiency, the present invention provides a kind of porcine pseudorabies virus, Porcine epidemic diarrhea virus vaccine combination, with
For the purpose of improving the body innate immunity, reach treatment PED purposes, the PRV of use and Caulis Spatholobi, matrimony vine, three taste Chinese medicine of Radix Codonopsis are equal
Play an important role of to improve the body innate immunity.Treatment formula is simple, and access approaches are extensive, of low cost, in PED is treated
With great promotional value.
To achieve these goals, the present invention adopts the following technical scheme that：
The preparation method of a kind of porcine pseudorabies virus, pig epidemic diarrhea vaccine combination, comprises the following steps：
Step 1, vaccine lysate is prepared
5-8g Caulis Spatholobis are taken, 6-10g matrimony vines after the mixing of 2-3g Radix Codonopsis, add in 30-40mL distilled water boilings 20-30 minutes,
After supernatant cooling, with 0.4 μm of membrane filtration, that is, vaccine lysate is made；
Step 2, mixed preparing vaccine combination
Take 2-3mL vaccines lysate dissolve 2-4 doses PRV inactivated vaccines, 2-4 doses PRV attenuated live vaccines,
2-3 doses PEDV inactivated vaccines, and be sufficiently mixed, that is, part vaccine combination is made.
A kind of porcine pseudorabies virus, pig epidemic diarrhea vaccine combination, part vaccine combination include 2-4 heads
Doses PRV inactivated vaccines, 2-4 doses PRV attenuated live vaccines, 2-3 doses PEDV inactivated vaccines and for dissolving
The vaccine lysate of above-mentioned three kinds of vaccines.
The present invention also provides this kind of porcine pseudorabies virus, pig epidemic diarrhea vaccine combination in treatment pig epidemic diarrhea
Application, pig epidemic diarrhea disease caused by being mainly used for emergency treatment PEDV takes 1/2 vaccine respectively for every pig
Composition is subcutaneously injected for neck and intramuscular injection.
The present invention compared with prior art, has the advantages that：
1st, vaccine lysate ingredient used is simple, crude drug source is wide, at low cost；PRV and PEDV vaccines are also readily available.
2nd, it is all very simple to dilute liquid and preparation method thereof and vaccine combination preparation method, can easily prepare, push away in onset site
Wide value is big.
Specific embodiment
With reference to embodiment, the invention will be further described, PRV inactivated vaccines in following embodiment, PRV live vaccines,
PEDV inactivated vaccines are bought from market.
With 6 pigs in each embodiment, wherein 4 for PED model foundations：With PEDV, (PEDV110, Chinese animal are defended
It is raw in epidemiology) Oral inoculation attacks poison, dosage is：5 age in days piglets, 2mL (104.5TCID50/mL)；15 age in days piglets,
0.5mL(105.5TCID50/mL)；25 age in days pigs, 2mL (105.5TCID50/mL)；Other 2 are used as control.
Judge curative effect：Excrement is normal in 2-3 days, to cure；Diarrhea mitigates, and is effective；It is invalid without effect is mitigated.
Embodiment 1 (it is young to attack malicious 5 ages in days；Symptom was apparent on 6th, came into effect treatment)：
(1) vaccine lysate is prepared：8g Caulis Spatholobis are weighed, 10g matrimony vines after the mixing of 3g Radix Codonopsis, add in 40mL distilled water and steam
It boils 20 minutes, after supernatant cooling, with 0.4 μm of membrane filtration, that is, vaccine lysate is made.
(2) vaccine combination is prepared：Composition manner of formulation is：2 doses PRV are dissolved with 2mL vaccines lysate to go out
Live vaccine, 2 doses PRV attenuated live vaccines, 2 doses PEDV inactivated vaccines, and be sufficiently mixed.
(3) application of vaccine combination：Piglet neck is subcutaneous and muscle injects 0.5mL compositions respectively.
Comparative example 1-1：It is same as Example 1, except substituting vaccine lysate with equivalent distilled water in step (1).
Comparative example 1-2：It is same as Example 1, do not contain PRV vaccines except preparing vaccine combination in step (2).
Comparative example 1-3：It is same as Example 1, except only carrying out intramuscular injection in step (3).
Embodiment 2 (it is young to attack malicious 15 ages in days；Symptom was apparent on 17th, came into effect treatment)：
(1) vaccine lysate is prepared：8g Caulis Spatholobis are weighed, 10g matrimony vines after the mixing of 3g Radix Codonopsis, add in 40mL distilled water and steam
It boils 20 minutes, after supernatant cooling, with 0.4 μm of membrane filtration, that is, vaccine lysate is made.
(2) vaccine combination is prepared：Composition manner of formulation is：2 doses PRV are dissolved with 2mL vaccines lysate to go out
Live vaccine, 2 doses PRV attenuated live vaccines, 2 doses PEDV inactivated vaccines, and be sufficiently mixed.
(3) application of vaccine combination：Piglet neck is subcutaneous and muscle injects 1.0mL compositions respectively.
Comparative example 2-1：It is same as Example 2, except substituting vaccine lysate with equivalent distilled water in step (1).
Comparative example 2-2：It is same as Example 2, do not contain PRV vaccines except preparing vaccine combination in step (2).
Comparative example 2-3：It is same as Example 1, except only carrying out intramuscular injection in step (3).
Embodiment 3 (it is young to attack malicious 25 ages in days；Symptom was apparent on 29th, came into effect treatment)：
(1) vaccine lysate is prepared：8g Caulis Spatholobis are weighed, 10g matrimony vines after the mixing of 3g Radix Codonopsis, add in 40mL distilled water and steam
It boils 20 minutes, after supernatant cooling, with 0.4 μm of membrane filtration, that is, vaccine lysate is made.
(2) vaccine combination is prepared：Composition manner of formulation is：4 doses PRV are dissolved with 3mL vaccines lysate to go out
Live vaccine, 4 doses PRV attenuated live vaccines, 3 doses PEDV inactivated vaccines, and be sufficiently mixed.
(3) application of vaccine combination：Piglet neck is subcutaneous and muscle injects 1.5mL compositions respectively.
Comparative example 3-1：It is same as Example 3, except substituting vaccine lysate with equivalent distilled water in step (1).
Comparative example 3-2：It is same as Example 3, do not contain PRV vaccines except preparing vaccine combination in step (2).
Comparative example 3-3：It is same as Example 3, except only carrying out intramuscular injection in step (3).
Table 1 (5 age in days)：It is all attack poison group piglet and connect poison all there is typical symptom of diarrhea after 1 day, show watery stools,
Inside there are white, yellow curdling block.Piglet spirit is extremely depressed, and hair is coarse, and feed intake is decreased obviously.
Poison is attacked to start to treat after 1 day
Note：Antibody detection method is：The PEDV antigens extracted through differential centrifugation are measured through ultraviolet absorption method, PEDV antigen bags
It is 3 μ g/mL by concentration, now during 37 DEG C of coating 2h, the phosphate buffer of 1.5%BSA puts 37 DEG C of closing 3h.The dilution factor of serum
For 1: 100.Pig enzyme labelled antibody extension rate is 1: 10000, reaction time 30min.OD450It is judged to the positive when >=0.3,0.5 >=
OD450>=0.3, be denoted as+；OD450>=0.5, be denoted as ++.
Table 2 (15 age in days)：It is all attack poison group piglet and connect poison all there is typical symptom of diarrhea after 2 days, show watery stools,
Inside there are white, yellow curdling block.Piglet spirit is extremely depressed, and hair is coarse, and feed intake is decreased obviously.
Poison is attacked to start to treat after 2 days
Note：Antibody detection method is：The PEDV antigens extracted through differential centrifugation are measured through ultraviolet absorption method, PEDV antigen bags
It is 3 μ g/mL by concentration, now during 37 DEG C of coating 2h, the phosphate buffer of 1.5%BSA puts 37 DEG C of closing 3h.The dilution factor of serum
For 1: 100.Pig enzyme labelled antibody extension rate is 1: 10000, reaction time 30min.OD450It is judged to the positive when >=0.3,0.5 >=
OD450>=0.3, be denoted as+；OD450>=0.5, be denoted as ++.
Table 3 (25 age in days)：It is all attack poison group piglet and connect poison all there is typical symptom of diarrhea after 2 days, show watery stools,
Inside there are white, yellow curdling block.Piglet spirit is extremely depressed, and hair is coarse, and feed intake is decreased obviously.
Poison is attacked to start to treat after 2 days
Note：Antibody detection method is：The PEDV antigens extracted through differential centrifugation are measured through ultraviolet absorption method, PEDV antigen bags
It is 3 μ g/mL by concentration, now during 37 DEG C of coating 2h, the phosphate buffer of 1.5%BSA puts 37 DEG C of closing 3h.The dilution factor of serum
For 1: 100.Pig enzyme labelled antibody extension rate is 1: 10000, reaction time 30min.OD450It is judged to the positive when >=0.3,0.5 >=
OD450>=0.3, be denoted as+；OD450>=0.5, be denoted as ++.
Claims (4)
1. a kind of preparation method of porcine pseudorabies virus, pig epidemic diarrhea vaccine combination, it is characterised in that：Including following step
Suddenly：
Step 1, vaccine lysate is prepared
5-8g Caulis Spatholobis are taken, 6-10g matrimony vines after the mixing of 2-3g Radix Codonopsis, add in 30-40mL distilled water boilings 20-30 minutes, supernatant
After liquid cooling, with 0.4 μm of membrane filtration, that is, vaccine lysate is made；
Step 2, vaccine combination is prepared
2-3mL vaccines lysate is taken to dissolve 2-4 doses PRV inactivated vaccines, 2-4 doses PRV attenuated live vaccines, 2-3
Head doses PEDV inactivated vaccines, and be sufficiently mixed, that is, part vaccine combination is made.
2. a kind of porcine pseudorabies virus, pig epidemic diarrhea vaccine group obtained by preparation method according to claim 1
Close object, it is characterised in that：One part vaccine combination includes 2-4 doses PRV inactivated vaccines, 2-4 doses PRV
Attenuated live vaccines, 2-3 doses PEDV inactivated vaccines and the vaccine lysate for dissolving above-mentioned three kinds of vaccines.
3. a kind of porcine pseudorabies virus according to claim 2, pig epidemic diarrhea vaccine combination are popular in treatment pig
The application of property diarrhea, it is characterised in that：The vaccine combination is used for pig epidemic diarrhea disease caused by emergency treatment PEDV.
4. a kind of porcine pseudorabies virus according to claim 3, pig epidemic diarrhea vaccine combination are popular in treatment pig
The application of property diarrhea, it is characterised in that：Subcutaneously being noted for neck for 1/2 part vaccine combination is taken respectively for every pig
It penetrates and for intramuscular injection.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101249260A (en) * | 2007-11-23 | 2008-08-27 | 河北师范大学 | Preparation of vaccines for preventing piglet diarrhoea |
| CN103070953A (en) * | 2012-12-28 | 2013-05-01 | 利川市五洲牧业有限责任公司 | Traditional Chinese medicine for curing epidemic diarrhea of pigs |
| CN103230574A (en) * | 2013-04-25 | 2013-08-07 | 中国农业科学院兰州畜牧与兽药研究所 | Traditional Chinese medical composition for treating porcine epizootic diarrhea and application thereof |
| CN104274511A (en) * | 2013-07-08 | 2015-01-14 | 洛阳惠中兽药有限公司 | Traditional Chinese medicinal composition for veterinary use and application thereof |
| CN104436142A (en) * | 2014-12-19 | 2015-03-25 | 何子双 | Traditional Chinese medicine composition for treating porcine epizootic diarrhea |
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- 2016-11-25 CN CN201611050420.6A patent/CN108096574A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101249260A (en) * | 2007-11-23 | 2008-08-27 | 河北师范大学 | Preparation of vaccines for preventing piglet diarrhoea |
| CN103070953A (en) * | 2012-12-28 | 2013-05-01 | 利川市五洲牧业有限责任公司 | Traditional Chinese medicine for curing epidemic diarrhea of pigs |
| CN103230574A (en) * | 2013-04-25 | 2013-08-07 | 中国农业科学院兰州畜牧与兽药研究所 | Traditional Chinese medical composition for treating porcine epizootic diarrhea and application thereof |
| CN104274511A (en) * | 2013-07-08 | 2015-01-14 | 洛阳惠中兽药有限公司 | Traditional Chinese medicinal composition for veterinary use and application thereof |
| CN104436142A (en) * | 2014-12-19 | 2015-03-25 | 何子双 | Traditional Chinese medicine composition for treating porcine epizootic diarrhea |
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| Title |
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