CN104162162A - Composition for resisting swine foot-and-mouth disease, freeze-dried powder, and preparation method and use of freeze-dried powder - Google Patents
Composition for resisting swine foot-and-mouth disease, freeze-dried powder, and preparation method and use of freeze-dried powder Download PDFInfo
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Abstract
The invention discloses a composition for resisting swine foot-and-mouth disease, freeze-dried powder, and a preparation method and use of the freeze-dried powder, and belongs to the technical field of veterinary feed additives. The composition comprises, by weight, 95-100 parts of a swine foot-and-mouth disease-resistant yolk antibody and 1-5 parts of a swine foot-and-mouth disease-resistant transfer factor. The freeze-dried powder comprises, by weight, 95-97 parts of the swine foot-and-mouth disease-resistant yolk antibody, 1-2 parts of the swine foot-and-mouth disease-resistant transfer factor, 0.2-0.3 parts of an inactivator, 0.01-0.02 parts of an antiseptic and 2.05-3.6 parts of a heat-resistant freeze-dried protective agent, and has the advantages of low preservation conditions, long preservation time, high antibody titer and good biological activity. Through combined use of the swine foot-and-mouth disease-resistant yolk antibody and the swine foot-and-mouth disease-resistant transfer factor, the freeze-dried powder has effects of preventing and treating swine foot-and-mouth disease, improves swine immunity and survival rate, is convenient for use and has a low cost.
Description
Technical field
The present invention is specifically related to anti-Schweineseuche compositions for a kind of feed additive, also relates to anti-Schweineseuche lyophilized powder and preparation method thereof and application simultaneously, belongs to field of feed additive technology for animals.
Background technology
Foot and mouth disease (Foot-and-mouth disease, FMD) be by foot and mouth disease virus (FMDV) cause Schweineseuche take infected animal mouth, there is the infectiousness epidemic disease that vesicle sexually transmitted disease (STD) disease is feature in hoof.The feature of foot and mouth disease is that morbidity is anxious, propagation is exceedingly fast.Except the solidity thing polluting by infection animal is propagated, can also aerocolloidal form grow propagation by air.Sickness rate can reach 100%, and piglet often loses symptom and dies suddenly, and when serious, mortality rate can reach 100%.Once this disease occurs, as has incured loss through delay in early days and put out, epidemic situation is normal to be expanded rapidly, causes ouster le main situation, and is difficult to eradicate.The effective measures of controlling and put out Schweineseuche are to slaughter disease pig and suspect contamination pig, and restriction animal is moved and meat listing, therefore causes serious economic loss.
In Zhu Yeshi China aquaculture, account for share heavier, 2009 are only 410.32 ten thousand tons of Henan Province's output Carnis Sus domesticas, it is quite large can releasing thus the annual product meat requirement of China, and be the trend that rises fast, but the disease occurring in pig industry is also diversified, the uninterrupted generation of Schweineseuche disease, all causes economic loss very to pig industry in recent years.
At present, people carry out immunity by vaccine to Schweineseuche disease, early stage results is better, but need to repeatedly inoculate, troublesome poeration, cost is higher, easily make poultry produce stress, and vaccine requires comparatively harsh for storage conditions, need cold preservation at low temperatures, and lower a period of time of room temperature condition lost efficacy.Therefore, in the urgent need to a kind of product that can prevent and treat Schweineseuche.
Summary of the invention
The object of this invention is to provide a kind of anti-Schweineseuche compositions.
Meanwhile, the present invention also provides a kind of anti-Schweineseuche lyophilized powder.
Moreover the present invention also provides a kind of preparation method of anti-Schweineseuche lyophilized powder.
Finally, the invention provides the application of a kind of anti-Schweineseuche lyophilized powder in preparing feedstuff.
In order to realize above object, the technical solution adopted in the present invention is:
An anti-Schweineseuche compositions, is comprised of the component of following weight portion: 95～100 parts of anti-Schweineseuche yolk antibodies, 1～5 part of anti-Schweineseuche transfer factor.
A kind of anti-Schweineseuche lyophilized powder; component by following weight portion forms: 95～97 parts of anti-Schweineseuche yolk antibodies, 1～2 part of anti-Schweineseuche transfer factor, 0.2～0.3 part of inactivator; 0.01～0.02 part of antiseptic, 2.05～3.6 parts of heat-resisting lyophilized protecting agents.
Described inactivator is formaldehyde.
Described antiseptic is thimerosal.
Preferably, described heat-resisting lyophilized protecting agent is comprised of the component of following weight portion: 0.05～0.1 part, selenium, 0.5～1 part of vitamin C, 1～2 part of polyvinylpyrrolidone, 0.5 part of glycine.
A preparation method for anti-Schweineseuche lyophilized powder, concrete steps are as follows:
(1) isolated strain is prepared vaccine: get that to make vaccine after swine foot-and-mouth disease virus strain propagation, deactivation standby;
(2) immune health chicken group prepares high-immunity egg: get vaccine virus immunization healthy chicken flock prepared by step (1), when antibody titer reaches 1:256, dry standby after the cleaning of collection high-immunity egg, sterilization;
(3) acidification: get the separated yolk of high-immunity egg prepared by step (2), add isopyknic sterilized water, be stirred well to the color yolk solution that turns white to obtain, in yolk solution, adding pH value is 4.9～5.2 pre-cooling acidifying water, addition is 8 times of yolk liquor capacity, mixes under rear room temperature and staticly settles;
(4) get the supernatant of step (3) after standing, by volume add 0.8% NaCl and NaSO
4mixture, to improve the degree of dissociation of antibody, centrifugal the supernatant that contains anti-Schweineseuche yolk antibody;
(5) get the supernatant in step (4), in supernatant, the amount of anti-Schweineseuche yolk antibody is 95～97 parts, then in supernatant, adds the anti-Schweineseuche transfer factor of 1～2 part, uses successively the membrane filtration of 0.45 μ m and 0.22 μ m after mixing;
(6) filtrate of step (5) is concentrated to obtain to concentrated solution; in concentrated solution, add successively the inactivator of 0.2～0.3 part, the antiseptic of 0.01～0.02 part and the heat-resisting lyophilized protecting agent of 2.05～3.6 parts, mix by pre-freeze, frozen dried and obtain anti-Schweineseuche lyophilized powder.
Described step is used inactivated vaccine normal amount in (2) for the first time, is 10
3.8eLD
50/ 0.1ml Schweineseuche inactivated vaccine 1.5mL, interval immunity in 7 days 1 time, follow-up each immunity increases by 1 times of amount, starts to detect antibody titer after immunity for the second time.
Anti-Schweineseuche transfer factor in described step (5) can be commercial goods, also can adopt following methods to prepare:
(1) with the normal saline after sterilizing, clean the pig spleen of crossing with swine foot-and-mouth disease virus antigen immune, remove the tissues such as its surperficial muscle, fat, then use normal saline flushing;
(2) by the pig spleen fragmentation after step (1) processing, add the PBS buffer of 2 times of pre-coolings, use tissue homogenate instrument by its homogeneous homogenate that obtains;
(3) homogenate in step (2) is being obtained to broken liquid with Ultrasonic Cell Disruptor processing 10min;
(4) the broken liquid of step (3) being adjusted to pH value is the centrifugal 30min of 5.3～5.7,6000rpm, collects supernatant;
(5), by the supernatant micro-filtrate membrane filtration of step (4), the daltonian material of molecular cut off 7000, collects supernatant;
(6) by the supernatant degerming membrane filtration of step (5), obtain anti-Schweineseuche transfer factor stock solution, 4 ℃ preserve or lyophilizing standby.
The content that the anti-Schweineseuche transfer factor adding in lyophilized powder of the present invention can be converted into stock solution transfer factor adds, and also can add the transfer factor powder through frozen dried.
The application of anti-Schweineseuche lyophilized powder in preparing feedstuff, step is as follows: getting anti-Schweineseuche lyophilized powder, is 0.2～0.6% to add in feedstuff according to percentage by weight, mixes and get final product.
Beneficial effect of the present invention:
Feed additive of the present invention merges two kinds of biological substances with different bioactive anti-Schweineseuches with anti-Schweineseuche lyophilized powder, can improve the ability of control Schweineseuche.Wherein, antibody is to have bioactive substance more targetedly, it is the bioactive substance that belongs to passive effect, can not effectively transfer the function of effective active of body, can only play a role corresponding virus, transfer factor is the active substance of a species specific adjusting immune function, can regulate the effective immune system function of body, thereby initiatively participate in immunity, stimulate the immunity of the body fluid interareas such as lymphocyte.Both couplings will improve attack and defense effect greatly, improve the function of preventing and treating of body.
The anti-Schweineseuche yolk antibody of the present invention can effectively prevent Schweineseuche, animal is not had to toxic and side effects, environmental protection, noresidue, the anti-Schweineseuche yolk antibody that height is exempted to make adds in feedstuff as adding component, can prevent largely Schweineseuche, strengthen addition and can also treat Schweineseuche disease; Anti-Schweineseuche shifts with anti-Schweineseuche yolk antibody and is used in conjunction with the prevention effect that can improve folk prescription.The advantages such as this lyophilized powder has reduction preservation condition, extends the holding time, and antibody titer is high, biological activity is good, it is added in feed formula by a certain percentage, can prevent and treat Schweineseuche disease, improve immunity and the survival rate of pig, easy to use, can reduce costs and labour force.
The present invention adopts healthy chicken flock as immune object, can reduce the probability of strain reversion, simultaneously can the high antibody titer of Quick to 1:256, outstanding to the sick prevention effect of Schweineseuche by the yolk antibody of making after chicken reinforced immunological, instant effect, evident in efficacy, and with strong points.
In the anti-Schweineseuche lyophilized powder of the present invention, heat-resisting lyophilized protecting agent adopts selenium, Vc, four kinds of components of polyvinylpyrrolidone (PVP) and glycine, wherein, selenium is a kind of trace element, tool antimutagenic effect, antioxidation, promote deactivation in carcinogenic body, cell proliferation effect, the effect of enhancing human body immunity power, regulate and improve the effect of body's immunity, make body specific immunity and nonspecific immunity, humoral immunization and cellular immune function are in relative balance state, there is anti-aging effects, can slow down the effective active of bioactive substance, the effect of Vc has a variety of, has non-oxidizability and avoids a series of variation after Exposure to Sunlight, can also regulate the adaptive capacity of variations in temperature to body, can also promote irony and calcareous Absorption simultaneously, PVP is adhesive, solubilizing agent, disintegrating agent, the prolongation of effect agent of medicine, utilize PVP molecule can play with hydrogen bond association between drug molecule and solubilising, quick-acting contrary prolongation of effect, slow releasing function, control PVP molecule and drug molecule between associating intensity, after can making to associate, drug molecule discharges lentamente in human body, delay its dissolution velocity, play prolongation drug effect and act on, there is obvious biological protection function simultaneously, anticorrosion, cushioning effect that glycine has, also can play the effect of stabilizing agent.
The specific embodiment
Following embodiment is only described in further detail the present invention, but does not form any limitation of the invention.
Embodiment 1
The anti-Schweineseuche compositions of the present embodiment, is comprised of the component of following weight portion: 97 parts of anti-Schweineseuche yolk antibodies, 1 part of anti-Schweineseuche transfer factor.
The anti-Schweineseuche lyophilized powder of the present embodiment, is comprised of the component of following weight portion: 97 parts of anti-Schweineseuche yolk antibodies, 1 part of anti-Schweineseuche transfer factor, 0.2 part, formaldehyde, 0.01 part of thimerosal, 0.05 part, selenium, 0.5 part of vitamin C, 1 part of polyvinylpyrrolidone, 0.5 part of glycine.
The preparation method of anti-Schweineseuche lyophilized powder in the present embodiment, concrete steps are as follows:
(1) strain is prepared vaccine: get that to make emulsifiable concentrate vaccine after national standard swine foot-and-mouth disease virus strain propagation, deactivation standby;
(2) immune health chicken group prepares high-immunity egg: getting emulsifiable concentrate vaccine virus immunization healthy chicken flock prepared by step (1), use for the first time inactivated vaccine normal amount, is 10
3.8eLD
50/ 0.1ml Schweineseuche inactivated vaccine 1.5mL, interval immunity in 7 days 1 time, follow-up each immunity increases by 1 times of amount, immunity is three times altogether, starts to detect antibody titer, after immunity for the second time when antibody titer reaches 1:256, start to collect high-immunity egg, high-immunity egg dries standby after cleaning, sterilization;
(3) acidification: get the separated yolk of high-immunity egg prepared by step (2), add isopyknic sterilized water, be stirred well to the color yolk solution that turns white to obtain, in yolk solution, adding pH value is 4.9 pre-cooling acidifying water, addition is 8 times of yolk liquor capacity, mixes to be placed in storage tank to staticly settle under room temperature;
(4) get the supernatant of step (3) after standing, by volume add 0.8% NaCl and NaSO
4(NaCl and NaSO
4mass ratio be 4:1) mixture, to improve the degree of dissociation of antibody, more centrifugal 20min must contain the supernatant of anti-Schweineseuche yolk antibody under 15000rpm/min;
(5) get the supernatant in step (4), in supernatant, the amount of anti-Schweineseuche yolk antibody is 97%, in supernatant, add again 1 part of anti-Schweineseuche transfer factor, mix rear use 0.45 μ m membrane filtration, with the molecular weight that dams, be greater than the macromolecular substances of 200KD, get filtrate with 0.22 μ m filter membrane secondary filter;
(6) filtrate of step (5) being concentrated into yolk antibody concentration through ultrafiltration apparatus is 2.5mL/500 plumage part, in concentrated solution, add successively 0.2 part of formaldehyde, 0.01 part of thimerosal, 0.05 part of selenium, 0.5 part of vitamin C, 1 part of polyvinylpyrrolidone, 0.5 part of glycine, mix and be placed at-80 ℃ fully freezingly, and be placed in lyophilized preparation lyophilizing and get final product.
In the present embodiment, the preparation method of anti-Schweineseuche transfer factor is:
(1) with the normal saline after sterilizing, clean the pig spleen of crossing with swine foot-and-mouth disease virus antigen immune, remove the tissues such as its surperficial muscle, fat, then use normal saline flushing;
(2) by the pig spleen fragmentation after step (1) processing, add the PBS buffer of 2 times of pre-coolings, use tissue homogenate instrument by its homogeneous homogenate that obtains;
(3) homogenate in step (2) is being obtained to broken liquid with Ultrasonic Cell Disruptor processing 10min;
(4) the broken liquid of step (3) being adjusted to pH value is the centrifugal 30min of 5.5,6000rpm, collects supernatant;
(5), by the supernatant micro-filtrate membrane filtration of step (4), the daltonian material of molecular cut off 7000, collects supernatant;
(6) by the supernatant degerming membrane filtration of step (5), obtain anti-Schweineseuche transfer factor stock solution, lyophilizing is standby.
The application of the anti-Schweineseuche lyophilized powder of the present embodiment in preparing feedstuff, step is as follows: getting anti-Schweineseuche lyophilized powder, is 0.2% to add in feedstuff according to percentage by weight, mixes and get final product.
Embodiment 2
The anti-Schweineseuche compositions of the present embodiment, is comprised of the component of following weight portion: 95 parts of anti-Schweineseuche yolk antibodies, 2 parts of anti-Schweineseuche transfer factors.
The anti-Schweineseuche lyophilized powder of the present embodiment, is comprised of the component of following weight portion: 95 parts of anti-Schweineseuche yolk antibodies, 2 parts of anti-Schweineseuche transfer factors, 0.3 part, formaldehyde, 0.02 part of thimerosal, 0.1 part, selenium, 1 part of vitamin C, 2 parts of polyvinylpyrrolidones, 0.5 part of glycine.
The preparation method of anti-Schweineseuche lyophilized powder in the present embodiment, concrete steps are as follows:
(1) strain is prepared vaccine: get that to make emulsifiable concentrate vaccine after national standard swine foot-and-mouth disease virus strain propagation, deactivation standby;
(2) immune health chicken group prepares high-immunity egg: getting emulsifiable concentrate vaccine virus immunization healthy chicken flock prepared by step (1), use for the first time inactivated vaccine normal amount, is 10
3.8eLD
50/ 0.1ml Schweineseuche inactivated vaccine 1.5mL, interval immunity in 7 days 1 time, follow-up each immunity increases by 1 times of amount, immunity is three times altogether, starts to detect antibody titer, after immunity for the second time when antibody titer reaches 1:256, start to collect high-immunity egg, high-immunity egg dries standby after cleaning, sterilization;
(3) acidification: get the separated yolk of high-immunity egg prepared by step (2), add isopyknic sterilized water, be stirred well to the color yolk solution that turns white to obtain, in yolk solution, adding pH value is 5.2 pre-cooling acidifying water, addition is 8 times of yolk liquor capacity, mixes to be placed in storage tank to staticly settle under room temperature;
(4) get the supernatant of step (3) after standing, by volume add 0.8% NaCl and NaSO
4(NaCl and NaSO
4mass ratio be 4:1) mixture, to improve the degree of dissociation of antibody, more centrifugal 20min must contain the supernatant of anti-Schweineseuche yolk antibody under 15000rpm/min;
(5) get the supernatant in step (4), in supernatant, the amount of anti-Schweineseuche yolk antibody is 95 parts, in supernatant, add again 2 parts of anti-Schweineseuche transfer factors (preparation method is with embodiment 1), mix rear use 0.45 μ m membrane filtration, with the molecular weight that dams, be greater than the macromolecular substances of 200KD, get filtrate with 0.22 μ m filter membrane secondary filter;
(6) filtrate of step (5) being concentrated into yolk antibody concentration through ultrafiltration apparatus is 2.5mL/500 plumage part, in concentrated solution, add successively 0.3 part of formaldehyde, 0.02 part of thimerosal, 0.1 part of selenium, 1 part of vitamin C, 2 parts of polyvinylpyrrolidones, 0.5 part of glycine, mix and be placed at-80 ℃ fully freezingly, and be placed in lyophilized preparation lyophilizing and get final product.
The application of the anti-Schweineseuche lyophilized powder of the present embodiment in preparing feedstuff, step is as follows: getting anti-Schweineseuche lyophilized powder, is 0.4% to add in feedstuff according to percentage by weight, mixes and get final product.
Embodiment 3
The anti-Schweineseuche compositions of the present embodiment, is comprised of the component of following weight portion: 96 parts of anti-Schweineseuche yolk antibodies, 1.6 parts of anti-Schweineseuche transfer factors.
The anti-Schweineseuche lyophilized powder of the present embodiment, component by following weight portion forms: 96 parts of anti-Schweineseuche yolk antibodies, 1.6 parts of anti-Schweineseuche transfer factors, 0.25 part, formaldehyde, 0.015 part of thimerosal, 0.075 part, selenium, 0.75 part of vitamin C, 1.5 parts of polyvinylpyrrolidones, 0.5 part of glycine.
The preparation method of anti-Schweineseuche lyophilized powder in the present embodiment, concrete steps are as follows:
(1) strain is prepared vaccine: get that to make emulsifiable concentrate vaccine after national standard swine foot-and-mouth disease virus strain propagation, deactivation standby;
(2) immune health chicken group prepares high-immunity egg: getting emulsifiable concentrate vaccine virus immunization healthy chicken flock prepared by step (1), use for the first time inactivated vaccine normal amount, is 10
3.8eLD
50/ 0.1ml Schweineseuche inactivated vaccine 1.5mL, interval immunity in 7 days 1 time, follow-up each immunity increases by 1 times of amount, immunity is three times altogether, starts to detect antibody titer, after immunity for the second time when antibody titer reaches 1:256, start to collect high-immunity egg, high-immunity egg dries standby after cleaning, sterilization;
(3) acidification: get the separated yolk of high-immunity egg prepared by step (2), add isopyknic sterilized water, be stirred well to the color yolk solution that turns white to obtain, in yolk solution, adding pH value is 5.0 pre-cooling acidifying water, addition is 8 times of yolk liquor capacity, mixes to be placed in storage tank to staticly settle under room temperature;
(4) get the supernatant of step (3) after standing, by volume add 0.8% NaCl and NaSO
4(NaCl and NaSO
4mass ratio be 4:1) mixture, to improve the degree of dissociation of antibody, more centrifugal 20min must contain the supernatant of anti-Schweineseuche yolk antibody under 15000rpm/min;
(5) get the supernatant in step (4), in supernatant, the amount of anti-Schweineseuche yolk antibody is 96 parts, in supernatant, add again 1.6 parts of anti-Schweineseuche transfer factors (preparation method is with embodiment 1), mix rear use 0.45 μ m membrane filtration, with the molecular weight that dams, be greater than the macromolecular substances of 200KD, get filtrate with 0.22 μ m filter membrane secondary filter;
(6) filtrate of step (5) being concentrated into yolk antibody concentration through ultrafiltration apparatus is 2.5mL/500 plumage part, in concentrated solution, add successively 0.25 part of formaldehyde, 0.015 part of thimerosal, 0.075 part of selenium, 0.75 part of vitamin C, 1.5 parts of polyvinylpyrrolidones, 0.5 part of glycine, mix and be placed at-80 ℃ fully freezingly, and be placed in lyophilized preparation lyophilizing and get final product.
The application of the anti-Schweineseuche lyophilized powder of the present embodiment in preparing feedstuff, step is as follows: getting anti-Schweineseuche lyophilized powder, is 0.6% to add in feedstuff according to percentage by weight, mixes and get final product.
Comparative example 1
Anti-Schweineseuche yolk antibody lyophilized powder for the feed additive of this comparative example, is comprised of the component of following weight portion: 97.6 parts of anti-Schweineseuche yolk antibodies, 0.25 part, formaldehyde, 0.015 part of thimerosal, 0.075 part, selenium, 0.75 part of vitamin C, 1.5 parts of polyvinylpyrrolidones, 0.5 part of glycine.The preparation method and application of lyophilized powder are with embodiment 3.
Comparative example 2
Anti-Schweineseuche transfer factor lyophilized powder for the feed additive of this comparative example, is comprised of the component of following weight portion: 97.6 parts of anti-Schweineseuche transfer factors, 0.25 part, formaldehyde, 0.015 part of thimerosal, 0.075 part, selenium, 0.75 part of vitamin C, 1.5 parts of polyvinylpyrrolidones, 0.5 part of glycine.
Test example 1
Experiment of preventive effects: use 2 the monthly age 700 of pigs, wherein 100 in contrast isolation support, use common feedstuff; Blank group of 100 conducts, adopt normal diet; Another 500 test isolated rearings, are divided into 5 groups at random, and 100 every group, the feedstuff that uses embodiment 1～3 and comparative example 1～2 to prepare, feed ingredient is consistent with matched group and blank group.Feeding manner is free choice feeding, freely drinks water, and after 24h, gets the rear FMD virus liquid of pig intramuscular injection inoculation dilution in test group and matched group, records 120h death condition, and result of the test refers to following table 1.
Therapeutic effect test: use 2 the monthly age 600 of pigs, the FMD virus liquid after 600 eye dripping inoculation dilutions, feeding manner is free choice feeding and freely drink water, after infection 12h, wherein 100 as positive controls, use normal diet; Another 500 is test group, is divided at random 5 groups, 100 every group, and the feedstuff that uses embodiment 1～3 and comparative example 1～2 to prepare, feed ingredient is consistent with positive controls, death condition of each group of continuous record 120h, result of the test refers to following table 1.
The result of each processed group control Schweineseuche of table 1
Conclusion: as shown in Table 1, the feedstuff of the embodiment of the present invention 1～3 preparation has good prophylactic-therapeutic effect to Schweineseuche, and protective rate and survival rate are all more than 87%, and the effect of its control Schweineseuche of the feedstuff of comparative example 1～2 preparation is not as embodiment.
Test example 2
The lyophilized powder of embodiment 1～3 and comparative example 1 preparation is kept in 37 ℃ of incubators, preserve 24 months, respectively at 1 week, 2 weeks after placing, January, February, April, August, December, 16 months, 20 months, 24 months, adopt agar diffusion method (AGP) to measure tiring of anti-Schweineseuche yolk antibody in each time point sample, testing result refers to following table 2.
37 ℃ of placement lyophilized powders of table 2 are in the variation of different time sections antibody titer
| Group/tiring | 1 week | 2 weeks | January | February | April | August | December | 16 months | 20 months | 24 months |
| Embodiment 1 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1:16 | 1:16 | 1:16 | 1:16 |
| Embodiment 2 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1:16 | 1:16 | 1:16 |
| Embodiment 3 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1:16 | 1:16 | 1:16 |
| Comparative example 1 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1:16 | 1:16 | 1:16 | 1:16 |
Conclusion, as shown in Table 2, the lyophilized powder storage life of the embodiment of the present invention 1～3 preparation is longer, is suitable for preserving under room temperature, and biological value time that changes is longer.
Quality standard
The quality standard of the lyophilized powder of embodiment 1～3 preparation refers to following several:
Character: faint yellow, be fine and close agglomerate.
Mycoplasma check: undertaken by < < Chinese veterinary pharmacopoeia > >, grow without mycoplasma.
Exogenous virus check: by < < Chinese veterinary pharmacopoeia > >, undertaken, up to specification.
Safety verification: with 5 of 7 Japanese instar chicklings, intramuscular injection 5 plumage parts, observe 2 weeks, all strong alive, injection site does not have pathological changes.
Residual moisture is measured: by < < Chinese veterinary pharmacopoeia > >, undertaken, meet veterinary biologics general rule regulation.
Shelf-life: according to room temperature bioactivity experimental result, can deposit 2 years.
Claims (10)
1. an anti-Schweineseuche compositions, is characterized in that: the component by following weight portion forms: 95～100 parts of anti-Schweineseuche yolk antibodies, 1～5 part of anti-Schweineseuche transfer factor.
2. an anti-Schweineseuche lyophilized powder; it is characterized in that: the component by following weight portion forms: 95～97 parts of anti-Schweineseuche yolk antibodies, 1～2 part of anti-Schweineseuche transfer factor, 0.2～0.3 part of inactivator; 0.01～0.02 part of antiseptic, 2.05～3.6 parts of heat-resisting lyophilized protecting agents.
3. anti-Schweineseuche lyophilized powder according to claim 2, is characterized in that: described inactivator is formaldehyde.
4. anti-Schweineseuche lyophilized powder according to claim 2, is characterized in that: described antiseptic is thimerosal.
5. anti-Schweineseuche lyophilized powder according to claim 2, is characterized in that: described heat-resisting lyophilized protecting agent is comprised of the component of following weight portion: 0.05～0.1 part, selenium, 0.5～1 part of vitamin C, 1～2 part of polyvinylpyrrolidone, 0.5 part of glycine.
6. a preparation method for the anti-Schweineseuche lyophilized powder as described in claim 2-5 any one, is characterized in that: concrete steps are as follows:
(1) isolated strain is prepared vaccine: get that to make vaccine after swine foot-and-mouth disease virus strain propagation, deactivation standby;
(2) immune health chicken group prepares high-immunity egg: get vaccine virus immunization healthy chicken flock prepared by step (1), when antibody titer reaches 1:256, dry standby after the cleaning of collection high-immunity egg, sterilization;
(3) acidification: get the separated yolk of high-immunity egg prepared by step (2), add isopyknic sterilized water, be stirred well to the color yolk solution that turns white to obtain, in yolk solution, adding pH value is 4.9～5.2 acidifying water, addition is 8 times of yolk liquor capacity, after mixing, staticly settles;
(4) get the supernatant of step (3) after standing, by volume add 0.8% NaCl and NaSO
4mixture, to improve the degree of dissociation of antibody, centrifugal the supernatant that contains anti-Schweineseuche yolk antibody;
(5) get the supernatant in step (4), 95～97 parts of the amounts of anti-Schweineseuche yolk antibody in supernatant, then in supernatant, add the anti-Schweineseuche transfer factor of 1～2 part, after mixing, use successively the membrane filtration of 0.45 μ m and 0.22 μ m;
(6) filtrate of step (5) is concentrated to obtain to concentrated solution; in concentrated solution, add successively the inactivator of 0.2～0.3 part, the antiseptic of 0.01～0.02 part and the heat-resisting lyophilized protecting agent of 2.05～3.6 parts, mix by pre-freeze, frozen dried and obtain anti-Schweineseuche lyophilized powder.
7. the preparation method of anti-Schweineseuche lyophilized powder according to claim 6, is characterized in that: described step is used inactivated vaccine normal amount in (2) for the first time, is 10
3.8eLD
50/ 0.1ml Schweineseuche inactivated vaccine 1.5mL, interval immunity in 7 days 1 time, follow-up each immunity increases by 1 times of amount, starts to detect antibody titer after immunity for the second time.
8. the preparation method of anti-Schweineseuche lyophilized powder according to claim 6, is characterized in that: NaCl and NaSO in described step (4)
4mass ratio be 4:1.
9. the preparation method of anti-Schweineseuche lyophilized powder according to claim 6, is characterized in that: the anti-Schweineseuche transfer factor in described step (5) adopts following methods to prepare:
(1) with the normal saline after sterilizing, clean the pig spleen of crossing with swine foot-and-mouth disease virus antigen immune, remove the tissues such as its surperficial muscle, fat, then use normal saline flushing;
(2) by the pig spleen fragmentation after step (1) processing, add the PBS buffer of 2 times of pre-coolings, use tissue homogenate instrument by its homogeneous homogenate that obtains;
(3) homogenate in step (2) is being obtained to broken liquid with Ultrasonic Cell Disruptor processing 10min;
(4) the broken liquid of step (3) being adjusted to pH value is the centrifugal 30min of 5.3～5.7,6000rpm, collects supernatant;
(5), by the supernatant micro-filtrate membrane filtration of step (4), the daltonian material of molecular cut off 7000, collects supernatant;
(6) by the supernatant degerming membrane filtration of step (5), obtain anti-Schweineseuche transfer factor stock solution, 4 ℃ preserve or lyophilizing standby.
10. the application of the anti-Schweineseuche lyophilized powder as described in claim 2-5 any one in preparing feedstuff, is characterized in that: step is as follows: getting anti-Schweineseuche lyophilized powder, is 0.2～0.6% to add in feedstuff according to percentage by weight, mixes and get final product.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104606675A (en) * | 2015-01-24 | 2015-05-13 | 西安天星生物药业股份有限公司 | Preparation method of poultry multi-linked specific egg yolk antibodies and transfer factors |
| CN105394392A (en) * | 2015-11-24 | 2016-03-16 | 天津市中升挑战生物科技有限公司 | Feed additive capable of resisting swine viral diseases and preparation method for feed additive |
| CN105906711A (en) * | 2016-05-03 | 2016-08-31 | 中农威特生物科技股份有限公司 | Preparation method, product and application of yolk antibody for resisting O-type and A-type foot-and-mouth disease viruses |
| CN106589117A (en) * | 2017-01-19 | 2017-04-26 | 天津市中升挑战生物科技有限公司 | Purified egg yolk antibody resisting to foot and mouth disease and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101259275A (en) * | 2008-04-22 | 2008-09-10 | 天津生机集团股份有限公司 | Yolk antibody feed additive and injection for resisting sheep virosis and preparation thereof |
| CN102716484A (en) * | 2012-05-31 | 2012-10-10 | 郑州后羿制药有限公司 | Duck virus hepatitis yolk antibody freeze-dried powder and preparation method thereof |
-
2013
- 2013-07-24 CN CN201310315447.3A patent/CN104162162B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101259275A (en) * | 2008-04-22 | 2008-09-10 | 天津生机集团股份有限公司 | Yolk antibody feed additive and injection for resisting sheep virosis and preparation thereof |
| CN102716484A (en) * | 2012-05-31 | 2012-10-10 | 郑州后羿制药有限公司 | Duck virus hepatitis yolk antibody freeze-dried powder and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 李文学 等: "犬脾脏转移因子的提取与鉴定", 《延边大学农学学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104606675A (en) * | 2015-01-24 | 2015-05-13 | 西安天星生物药业股份有限公司 | Preparation method of poultry multi-linked specific egg yolk antibodies and transfer factors |
| CN104606675B (en) * | 2015-01-24 | 2019-08-30 | 深圳金石邦信股权投资基金管理有限公司 | A kind of multi-joint special yolk antibody of fowl and the preparation method of transfer factor |
| CN105394392A (en) * | 2015-11-24 | 2016-03-16 | 天津市中升挑战生物科技有限公司 | Feed additive capable of resisting swine viral diseases and preparation method for feed additive |
| CN105906711A (en) * | 2016-05-03 | 2016-08-31 | 中农威特生物科技股份有限公司 | Preparation method, product and application of yolk antibody for resisting O-type and A-type foot-and-mouth disease viruses |
| CN105906711B (en) * | 2016-05-03 | 2019-09-27 | 中农威特生物科技股份有限公司 | The preparation method of a kind of resisting O-type and A type foot and mouth disease virus Yolk antibody, product and application thereof |
| CN106589117A (en) * | 2017-01-19 | 2017-04-26 | 天津市中升挑战生物科技有限公司 | Purified egg yolk antibody resisting to foot and mouth disease and preparation method thereof |
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