CN104208675A - Mixed freeze-dried powder for preventing chicken viral diseases and preparation method thereof - Google Patents
Mixed freeze-dried powder for preventing chicken viral diseases and preparation method thereof Download PDFInfo
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Abstract
The invention discloses mixed freeze-dried powder for preventing chicken viral diseases and a preparation method thereof. The mixed freeze-dried powder comprises the following components in parts by weight: 90 to 95 parts of chicken viral disease preventing yolk antibody, 1 to 2 parts f chicken viral disease preventing transfer factor, 1 to 2 parts of alpha-interferon, 1 to 2 parts of antioxidant, 0.01 to 0.02 part of antiseptic, 0.2 to 0.3 parts of inactivating agent, 0.05 to 0.1 part of selenium, 1 to 2 parts of polyvinylpyrrolidone, 0.5 to 1 part of glycine, and 0.5 to 1 part of active lactobacillus. The provided mixed freeze-dried powder for preventing chicken viral diseases is prepared by compounding chicken viral disease preventing yolk antibody, chicken viral disease preventing transfer factor, and an immunity enhancer, moreover a plurality of auxiliary materials are added, the raw materials cooperate with each other to generate a synergetic effect, the effect on preventing and treating chicken viral diseases is prominently improved, and the freeze-dried powder has the advantages of strong pertinency, long preservation time at a normal temperature, high antibody titer, and good bio-activity.
Description
Technical field
The invention belongs to veterinary drug technical field, relate to a kind of freeze-dried mixed powder of anti-chicken viral diseases, also relate to a kind of preparation method of freeze-dried mixed powder of anti-chicken viral diseases simultaneously.
Background technology
Chicken viral diseases is referred to as the one that is acute, hyperinfection disease in various degree caused by different virus, comprises the diseases such as newcastle disease, infectious bronchitis, bird flu.Chicken viral diseases is modal disease in a kind of poultry husbandry, and because it has, morbidity is unexpected, the course of disease is short, mortality rate high, at present one of main harm remaining poultry husbandry.
The method of current control chicken viral diseases is more, mainly uses Western medicine, immunoprophylaxis, Chinese herbal medicine etc. to prevent and treat.Wherein, spreading unchecked that Western medicine uses brings great potential safety hazard to Carnis Gallus domesticus based food, and drug residue has become one of principal element affecting chicken meat product safety.Immunoprophylaxis early stage results is better, but needs repeatedly to inoculate, troublesome poeration, and cost is higher, easily causes stress; And vaccine requires higher for storage conditions, need cold preservation under 2-8 DEG C of condition, room temperature condition lower a period of time then loses effect.Chinese herbal feed additive is also one of current research direction, but because the kind of Chinese herbal medicine is more, identical Chinese crude drug is very large to the difference between the effects of different diseases of bird and livestock, and Chinese herbal medicine is extremely limited to the immune effect of disease, therefore applies and also has certain limitation.
Summary of the invention
The object of this invention is to provide a kind of freeze-dried mixed powder of anti-chicken viral diseases, solve the operation inconvenience that prior art control chicken viral diseases exists, the problem that medicament is not easily preserved, effect is poor.
Second object of the present invention is to provide a kind of preparation method of freeze-dried mixed powder of anti-chicken viral diseases.
In order to realize above object, the technical solution adopted in the present invention is: a kind of freeze-dried mixed powder of anti-chicken viral diseases, comprises the component of following parts by weight: anti-chicken virus yolk antibody 90 ~ 95 parts, anti-chicken virus transfer factor 1 ~ 2 part, alpha-interferon 1 ~ 2 part, antioxidant 1 ~ 2 part, antiseptic 0.01 ~ 0.02 part, inactivator 0.2 ~ 0.3 part, 0.05 ~ 0.1 part, selenium, polyvinylpyrrolidone 1 ~ 2 part, glycine 0.5 ~ 1 part, active lactobacillus 0.5 ~ 1 part.
Described inactivator is formaldehyde.
Described antiseptic is thimerosal.
Described antioxidant be in vitamin C, vitamin E any one or its combination.
A preparation method for the freeze-dried mixed powder of above-mentioned anti-chicken viral diseases, comprises the following steps:
A. adopt the strain of chicken viral diseases Virus Standard to prepare vaccine, immune health chicken group obtains high-immunity egg, by the supernatant that high-immunity egg obtains containing anti-chicken virus yolk antibody through sterilization, acidification, centrifugal treating;
B. the supernatant containing anti-chicken virus yolk antibody is mixed with anti-chicken virus transfer factor, antiseptic, inactivator, selenium, antioxidant, alpha-interferon, active lactobacillus, polyvinylpyrrolidone, glycine, then lyophilizing and get final product.
Preferably, the preparation method of the freeze-dried mixed powder of above-mentioned anti-chicken viral diseases, comprises the following steps:
1) get the strain of chicken viral diseases Virus Standard to breed, its deactivation is prepared into vaccine, at interval of 7 days, immunity is carried out to healthy laying hen group, immunity three times altogether, start to detect antibody titer after second time immunity, when antibody titer reaches 1:256, collect high-immunity egg, the high-immunity egg of collection is carried out clear up, sterilize, dry for subsequent use;
2) separation of step 1) gained high-immunity egg is obtained high-immunity egg yolk, add isopyknic sterilized water, fully stir, obtain yolk antibody solution, in yolk antibody solution, add acidified aqueous solution, mix, staticly settle, get supernatant for subsequent use;
3) in step 2) add sodium salt in gained supernatant and make sodium salt volume account for 0.8% of cumulative volume, centrifugalize, must supernatant containing anti-chicken virus yolk antibody;
4) get the supernatant of the anti-chicken virus yolk antibody containing 90 ~ 95 weight portions of step 3) gained, add the anti-chicken virus transfer factor of 1 ~ 2 weight portion, filter, reserved filtrate is for subsequent use;
5) step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, add antiseptic 0.01 ~ 0.02 weight portion, inactivator 0.2 ~ 0.3 weight portion, selenium 0.05 ~ 0.1 weight portion, antioxidant 1 ~ 2 weight portion, alpha-interferon 1 ~ 2 weight portion, active lactobacillus 0.5 ~ 1 weight portion, polyvinylpyrrolidone 1 ~ 2 weight portion, glycine 0.5 ~ 1 weight portion mix, first-20 DEG C of pre-coolings, lyophilizing under-80 DEG C of conditions, to obtain final product again.
Step 2) described in the pH of acidified aqueous solution be 4.9 ~ 5.2; The addition of acidified aqueous solution is 8 times of yolk antibody liquor capacity.Described acidified aqueous solution can be acidified aqueous solution.
Sodium salt described in step 3) is NaCl and NaSO
4mass ratio be the mixture of 4:1.
The rotating speed of centrifugalize described in step 3) is 15000rpm, and the time is 20min.
Filtering described in step 4) is first filter with the filter membrane of 0.45 μm, and molecular cut off is greater than the macromole of 200KD, then by the membrane filtration of filtrate with 0.22 μm.
Yolk antibody is a kind of immunoglobulin, due to body is subject to the stimulation of external antigen and the one that produces stress product, can combine with corresponding antigens, eliminate the cause of disease.The present invention, by the chicken group of the vaccine virus immunization health of chicken viral diseases, can make to produce effective anti-chicken viral yolk antibody in the egg yolk of acquisition, decrease the probability of strain reversion simultaneously.
Transfer factor carries the specific immunity information of primed lymphocyte, can by specific immunity information in passing recipient lymphocytes, the lymphocyte of receptor non-activity is made to change specificity primed lymphocyte into, thus the immunoreation exciting recipient cell to mediate.
Alpha-interferon makes cell produce antiviral protein by cell surface receptor effect, also can strengthen natural killer cell (NK cell), macrophage and the lymphocytic vigor of T simultaneously, thus play immunoregulation effect, and strengthen anti-virus ability.
In the freeze-dried mixed powder of anti-chicken viral diseases of the present invention, anti-chicken virus yolk antibody is a kind of immunoglobulin, has good prevention effect to chicken viral diseases; Anti-chicken virus transfer factor itself is a kind of special antiviral agent; Alpha-interferon, active lactobacillus can the function of immune system of enhancing body as immunostimulant, are combined by above-mentioned substance, greatly can improve the effect of prevention and therapy chicken viral diseases.
The freeze-dried mixed powder of anti-chicken viral diseases of the present invention, by anti-chicken virus yolk antibody, anti-chicken virus transfer factor and immunostimulant (alpha-interferon, active lactobacillus) carry out composite, and be aided with antioxidant, antiseptic, inactivator, selenium, freeze-dried mixed powder prepared by polyvinylpyrrolidone and glycine, each component cooperatively interacts, coordinative role, the effect of its control chicken viral diseases can be significantly improved, by force pointed, the room temperature lower holding time is long, antibody titer is high, the advantage that biological activity is good, it is a kind of potent biological preparation with wide application prospect, prove through animal experiment, the freeze-dried mixed powder of anti-chicken viral diseases of the present invention, obviously will to be better than in anti-chicken virus yolk antibody, anti-chicken virus transfer factor, alpha-interferon any one to the prevention effect of chicken viral diseases and to be used alone or wherein two kinds of situations about combinationally using.
The freeze-dried mixed powder of anti-chicken viral diseases of the present invention, during use, can be used as feed additive and is mixed in feedstuff by mass fraction 2% ~ 3%; Adopt the administration of intramuscular injection mode during morbidity, containing freeze-dried mixed powder 1g in every 100ml injection, the use amount of injection is each 0.1 ~ 0.2ml/kg body weight; Medication first doubles, and therapeutic effect is better.
The preparation method of the freeze-dried mixed powder of anti-chicken viral diseases of the present invention, anti-chicken virus yolk antibody, anti-chicken virus transfer factor, immunostimulant (alpha-interferon, active lactobacillus) and multiple auxiliary materials are carried out composite, and by adjustment adjuvant composition, control its consumption, extend the holding time of yolk antibody further, ensure that the activity in the yolk antibody long period at normal temperatures, reduce the storage and transport cost of freeze-dried powder preparation, easy to use, be with a wide range of applications; Preparation method technique is simple, easy to operate, greatly reduces production cost, is applicable to large-scale industrial production.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the present invention is further illustrated.
In the specific embodiment of the present invention, fowl disease poison strain used is national standard strain; Alpha-interferon used is purchased from Changchun Biological Products Institute.
In the specific embodiment of the present invention, anti-chicken virus transfer factor used, prepared by following methods:
1), after immunity being carried out to pig spleen and/or thymus with chicken viral diseases standard virus antigen, its surperficial muscle, fat is removed, then with the normal saline flushing after sterilizing;
2) by after the pig spleen after flushing and/or thymus fragmentation, the PBS buffer after the pre-cooling of 2 times of volumes is added, with tissue homogenate instrument, it is homogeneous, obtain homogenate;
3) by homogenate with after Ultrasonic Cell Disruptor process 5 ~ 10min, regulate pH be 5.5, centrifugal 30min under 5000 ~ 6000rpm rotating speed, collect supernatant;
4) by step 3) gained supernatant micro-filtrate membrane filtration, molecular cut off is the material of more than 7000 dalton, and gained filtrate is used degerming membrane filtration, obtains anti-chicken virus transfer factor stock solution, saves backup under 4 DEG C of conditions.
Embodiment 1
The freeze-dried mixed powder of the anti-chicken viral diseases of the present embodiment is the freeze-dried mixed powder of anti-infectious bursal disease, comprises the component of following parts by weight: anti-infectious bursal disease yolk antibody 90 parts, anti-infectious bursal disease transfer factor 2 parts, vitamin C 1 part, vitamin e1 part, alpha-interferon 2 parts, active lactobacillus 1 part, thimerosal 0.02 part, 0.2 part, formaldehyde, 0.05 part, selenium, polyvinylpyrrolidone 2 parts, glycine 0.5 part.
The preparation method of the freeze-dried mixed powder of the anti-infectious bursal disease of the present embodiment, comprises the following steps:
1) get chicken infectivity bursa of Fabricius virus standard strain (B87) to breed, its deactivation is prepared into vaccine, carried out immunity at interval of 7 days to healthy laying hen group, altogether immunity three times, first time, immune health chicken group inoculated inactivated vaccine 1.5ml, for the second time, third time immunity inoculation dosage 1 times, start to detect antibody titer after second time immunity, when antibody titer reaches 1:256, collect high-immunity egg, the high-immunity egg of collection is carried out clear up, sterilize, dry for subsequent use;
2) separation of step 1) gained high-immunity egg is obtained high-immunity egg yolk, add isopyknic sterilized water, abundant stirring, obtain yolk antibody solution, the acidified aqueous solution that pH is 5.0 is added in yolk antibody solution, acidified aqueous solution addition is 8 times of yolk antibody liquor capacity, mixes, staticly settles, get supernatant for subsequent use;
3) in step 2) add by NaCl and NaSO in gained supernatant
4be the mixing sodium salt that the ratio of 4:1 is mixed to form in mass ratio, make mixing sodium salt volume account for 0.8% of cumulative volume, with the centrifugal 20min of the rotating speed of 15000rpm, the supernatant of anti-infectious bursal disease yolk antibody must be contained;
4) supernatant of the anti-infectious bursal disease yolk antibody containing 90 weight portions of step 3) gained is got, add the anti-infectious bursal disease transfer factor of 2 weight portion, filter with the filter membrane of 0.45 μm, the molecular weight that dams is greater than the macromolecular substances of 200KD, again by filtrate through 0.22 μm of membrane filtration, reserved filtrate is for subsequent use;
5) step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, add thimerosal 0.02 weight portion, formaldehyde 0.2 weight portion, selenium 0.05 weight portion, vitamin C 1 weight portion, vitamin e1 weight portion, active lactobacillus 1 weight portion, alpha-interferon 2 weight portion, polyvinylpyrrolidone 2 weight portion, glycine 0.5 weight portion mix, first-20 DEG C of pre-coolings, lyophilizing under-80 DEG C of conditions, to obtain final product again.
Embodiment 2
The freeze-dried mixed powder of the anti-chicken viral diseases of the present embodiment is the freeze-dried mixed powder of anti-infectious bronchitis of chicken, comprises the component of following parts by weight: anti-infectious bronchitis of chicken yolk antibody 93 parts, anti-infectious bronchitis of chicken transfer factor 1.5 parts, vitamin C 0.5 part, vitamin E 0.5 part, alpha-interferon 1.5 parts, active lactobacillus 1 part, thimerosal 0.01 part, 0.3 part, formaldehyde, 0.1 part, selenium, polyvinylpyrrolidone 1.5 parts, glycine 1 part.
The preparation method of the freeze-dried mixed powder of the anti-infectious bronchitis of chicken of the present embodiment, comprises the following steps:
1) get avian infectious bronchitis virus standard strain (M41) to breed, its deactivation is prepared into vaccine, carried out immunity at interval of 7 days to healthy laying hen group, altogether immunity three times, first time, immune health chicken group inoculated inactivated vaccine 1.5ml, for the second time, third time immunity inoculation dosage 1 times, start to detect antibody titer after second time immunity, when antibody titer reaches 1:256, collect high-immunity egg, the high-immunity egg of collection is carried out clear up, sterilize, dry for subsequent use;
2) separation of step 1) gained high-immunity egg is obtained high-immunity egg yolk, add isopyknic sterilized water, abundant stirring, obtain yolk antibody solution, the acidified aqueous solution that pH is 5.0 is added in yolk antibody solution, acidified aqueous solution addition is 8 times of yolk antibody liquor capacity, mixes, staticly settles, get supernatant for subsequent use;
3) in step 2) add by NaCl and NaSO in gained supernatant
4be the mixing sodium salt that the ratio of 4:1 is mixed to form in mass ratio, make mixing sodium salt volume account for 0.8% of cumulative volume, with the centrifugal 20min of the rotating speed of 15000rpm, the supernatant of anti-infectious bronchitis of chicken yolk antibody must be contained;
4) supernatant of the anti-infectious bronchitis of chicken yolk antibody containing 93 weight portions of step 3) gained is got, add the anti-infectious bronchitis of chicken transfer factor of 1.5 weight portion, filter with the filter membrane of 0.45 μm, the molecular weight that dams is greater than the macromolecular substances of 200KD, again by filtrate through 0.22 μm of membrane filtration, reserved filtrate is for subsequent use;
5) step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, add thimerosal 0.01 weight portion, formaldehyde 0.3 weight portion, selenium 0.1 weight portion, vitamin C 0.5 weight portion, vitamin E 0.5 weight portion, active lactobacillus 1 weight portion, alpha-interferon 1.5 weight portion, polyvinylpyrrolidone 1.5 weight portion, glycine 1 weight portion mix, first-20 DEG C of pre-coolings, lyophilizing under-80 DEG C of conditions, to obtain final product again.
Embodiment 3
The freeze-dried mixed powder of the anti-chicken viral diseases of the present embodiment is the freeze-dried mixed powder of anti-avian influenza, comprises the component of following parts by weight: anti-avian influenza yolk antibody 95 parts, anti-avian influenza transfer factor 1 part, vitamin C 0.5 part, vitamin E 0.5 part, alpha-interferon 1 part, active lactobacillus 0.5 part, thimerosal 0.01 part, 0.25 part, formaldehyde, 0.08 part, selenium, polyvinylpyrrolidone 1 part, glycine 0.5 part.
The preparation method of the freeze-dried mixed powder of the anti-infectious bronchitis of chicken of the present embodiment, comprises the following steps:
1) get bird flu virus (Hp) standard strain to breed, its deactivation is prepared into vaccine, carried out immunity at interval of 7 days to healthy laying hen group, altogether immunity three times, first time, immune health chicken group inoculated inactivated vaccine 1.5ml, for the second time, third time immunity inoculation dosage 1 times, start to detect antibody titer after second time immunity, when antibody titer reaches 1:256, collect high-immunity egg, the high-immunity egg of collection is carried out clear up, sterilize, dry for subsequent use;
2) separation of step 1) gained high-immunity egg is obtained high-immunity egg yolk, add isopyknic sterilized water, abundant stirring, obtain yolk antibody solution, the acidified aqueous solution that pH is 5.0 is added in yolk antibody solution, acidified aqueous solution addition is 8 times of yolk antibody liquor capacity, mixes, staticly settles, get supernatant for subsequent use;
3) in step 2) add by NaCl and NaSO in gained supernatant
4be the mixing sodium salt that the ratio of 4:1 is mixed to form in mass ratio, make mixing sodium salt volume account for 0.8% of cumulative volume, with the centrifugal 20min of the rotating speed of 15000rpm, the supernatant of anti-avian influenza yolk antibody must be contained;
4) supernatant of the anti-avian influenza yolk antibody containing 95 weight portions of step 3) gained is got, add 1 weight portion anti-avian influenza transfer factor, filter with the filter membrane of 0.45 μm, the molecular weight that dams is greater than the macromolecular substances of 200KD, again by filtrate through 0.22 μm of membrane filtration, reserved filtrate is for subsequent use;
5) step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, add thimerosal 0.01 weight portion, formaldehyde 0.25 weight portion, selenium 0.08 weight portion, vitamin C 0.5 weight portion, vitamin E 0.5 weight portion, active lactobacillus 0.5 weight portion, alpha-interferon 1 weight portion, polyvinylpyrrolidone 1 weight portion, glycine 0.5 weight portion mix, first-20 DEG C of pre-coolings, lyophilizing under-80 DEG C of conditions, to obtain final product again.
Experimental example 1
This experimental example carries out Performance Detection to the freeze-dried mixed powder of embodiment 1 ~ 3 gained, and result is as shown in table 1,2.
Room temperature bioactivity experimental technique: the freeze-dried mixed powder preparation of embodiment 1 ~ 3 gained is kept in 37 DEG C of incubators, preserve 24 months, respectively 1 week, 2 weeks, January, February, April, August, December, after 16 months, 20 months, 24 months, by agar diffusion (AGP) method, the yolk antibody measuring each time point institute sample thief is tired change.
Table 1 embodiment 1 ~ 3 gained freeze-dried mixed powder room temperature bioactivity experimental result
| Group | 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 |
The freeze-dried mixed powder room temperature of table 2 embodiment 1 ~ 3 gained places performance test results
Experimental example 2
The effect of this experimental example to the freeze-dried mixed powder control infectious bursal disease of the anti-infectious bursal disease of embodiment 1 carries out clinical experiment.
Experimental technique: choose 1 the monthly age chicken 900, wherein 100 as the isolated rearing of counteracting toxic substances group, 100 as the isolated rearing of blank group, 600 are divided into 6 groups as a control group, often organize 100, isolated rearing, uses the preparation for treating of comparative example 1 ~ 6 respectively, other 100 as test group, with the preparation for treating of embodiment 1.Feeding manner is free choice feeding normal diet, freely drinks water, and after 24h, get the chicken infectivity bursa of Fabricius virus liquid after the chicken intramuscular injection inoculation dilution of test group, matched group and counteracting toxic substances group, inoculum concentration is 2ml/; Start immunotherapy targeted autoantibody after infection, intramuscular injection, injection dosage is 0.2ml/kg body weight; Death condition after record 120h, result is as shown in table 3.
The freeze-dried mixed powder of table 3 embodiment 1 gained is to the prevention effect experimental result of infectious bursal disease
Wherein, the reagent combination of comparative example 1 ~ 6 is as shown in table 4.
The reagent combination of table 4 comparative example 1 ~ 6
As can be seen from table 3,4, the freeze-dried mixed powder of embodiment 1 gained effectively can prevent and treat infections chicken cloacal bursa, and freeze-dried mixed powder effect obviously will to be better than in anti-infectious bursal disease yolk antibody, anti-infectious bursal disease transfer factor, alpha-interferon any one is used alone or wherein two kinds of situations about combinationally using.
Experimental example 3
The effect of this experimental example to the freeze-dried mixed powder control infectious bronchitis of chicken of the anti-infectious bronchitis of chicken of embodiment 2 carries out clinical experiment.
Experimental technique: choose 1 the monthly age chicken 900, wherein 100 as the isolated rearing of counteracting toxic substances group, 100 as the isolated rearing of blank group, 600 are divided into 6 groups as a control group, often organize 100, isolated rearing, uses the preparation for treating of comparative example 7 ~ 12 respectively, other 100 as test group, with the preparation for treating of embodiment 2.Feeding manner is free choice feeding normal diet, freely drinks water, and after 24h, get the avian infectious bronchitis virus liquid after the chicken intramuscular injection inoculation dilution of test group, matched group and counteracting toxic substances group, inoculum concentration is 2ml/; Start immunotherapy targeted autoantibody after infection, intramuscular injection, injection dosage is 0.2ml/kg body weight; Death condition after record 120h, result is as shown in table 5.
The freeze-dried mixed powder of table 5 embodiment 2 gained is to the prevention effect experimental result of infectious bronchitis of chicken
Wherein, the reagent combination of comparative example 7 ~ 12 is as shown in table 6.
The reagent combination of table 6 comparative example 7 ~ 12
As can be seen from table 5,6, the freeze-dried mixed powder of embodiment 2 gained effectively can prevent and treat infectious bronchitis of chicken, and freeze-dried mixed powder effect obviously will to be better than in anti-infectious bronchitis of chicken yolk antibody, anti-infectious bronchitis of chicken transfer factor, alpha-interferon any one is used alone or wherein two kinds of situations about combinationally using.
Experimental example 4
The effect of this experimental example to the freeze-dried mixed powder control bird flu of the anti-avian influenza of embodiment 3 carries out clinical experiment.
Experimental technique: choose 1 the monthly age chicken 900, wherein 100 as the isolated rearing of counteracting toxic substances group, 100 as the isolated rearing of blank group, 600 are divided into 6 groups as a control group, often organize 100, isolated rearing, uses the preparation for treating of comparative example 1 ~ 6 respectively, other 100 as test group, with the preparation for treating of embodiment 3.Feeding manner is free choice feeding normal diet, freely drinks water, and after 24h, get the avian influenza venom after the chicken intramuscular injection inoculation dilution of test group, matched group and counteracting toxic substances group, inoculum concentration is 2ml/; Start immunotherapy targeted autoantibody after infection, intramuscular injection, injection dosage is 0.2ml/kg body weight; Death condition after record 120h, result is as shown in table 3.
The freeze-dried mixed powder of table 7 embodiment 7 gained is to the prevention effect experimental result of bird flu
Wherein, the reagent combination of comparative example 12 ~ 18 is as shown in table 8.
The reagent combination of table 8 comparative example 12 ~ 18
As can be seen from table 7,8, the freeze-dried mixed powder of embodiment 3 gained effectively can prevent and treat bird flu, and freeze-dried mixed powder effect obviously will to be better than in anti-avian influenza yolk antibody, anti-avian influenza transfer factor, alpha-interferon any one is used alone or wherein two kinds of situations about combinationally using.
Claims (10)
1. a freeze-dried mixed powder for anti-chicken viral diseases, is characterized in that: the component comprising following parts by weight: anti-chicken virus yolk antibody 90 ~ 95 parts, anti-chicken virus transfer factor 1 ~ 2 part, alpha-interferon 1 ~ 2 part, antioxidant 1 ~ 2 part, antiseptic 0.01 ~ 0.02 part, inactivator 0.2 ~ 0.3 part, 0.05 ~ 0.1 part, selenium, polyvinylpyrrolidone 1 ~ 2 part, glycine 0.5 ~ 1 part, active lactobacillus 0.5 ~ 1 part.
2. the freeze-dried mixed powder of anti-chicken viral diseases according to claim 1, is characterized in that: described inactivator is formaldehyde.
3. the freeze-dried mixed powder of anti-chicken viral diseases according to claim 1, is characterized in that: described antiseptic is thimerosal.
4. the freeze-dried mixed powder of anti-chicken viral diseases according to claim 1, is characterized in that: described antioxidant be in vitamin C, vitamin E any one or its combination.
5. a preparation method for the freeze-dried mixed powder of anti-chicken viral diseases as claimed in claim 1, is characterized in that: comprise the following steps:
A. adopt the strain of chicken viral diseases Virus Standard to prepare vaccine, immune health chicken group obtains high-immunity egg, by the supernatant that high-immunity egg obtains containing anti-chicken virus yolk antibody through sterilization, acidification, centrifugal treating;
B. the supernatant containing anti-chicken virus yolk antibody is mixed with anti-chicken virus transfer factor, antiseptic, inactivator, selenium, antioxidant, alpha-interferon, active lactobacillus, polyvinylpyrrolidone, glycine, then lyophilizing and get final product.
6. the preparation method of the freeze-dried mixed powder of anti-chicken viral diseases according to claim 5, is characterized in that: comprise the following steps:
1) get the strain of chicken viral diseases Virus Standard to breed, its deactivation is prepared into vaccine, at interval of 7 days, immunity is carried out to healthy laying hen group, immunity three times altogether, start to detect antibody titer after second time immunity, when antibody titer reaches 1:256, collect high-immunity egg, the high-immunity egg of collection is carried out clear up, sterilize, dry for subsequent use;
2) separation of step 1) gained high-immunity egg is obtained high-immunity egg yolk, add isopyknic sterilized water, fully stir, obtain yolk antibody solution, in yolk antibody solution, add acidified aqueous solution, mix, staticly settle, get supernatant for subsequent use;
3) in step 2) add sodium salt in gained supernatant and make sodium salt volume account for 0.8% of cumulative volume, centrifugalize, must supernatant containing anti-chicken virus yolk antibody;
4) get the supernatant of the anti-chicken virus yolk antibody containing 90 ~ 95 weight portions of step 3) gained, add the anti-chicken virus transfer factor of 1 ~ 2 weight portion, filter, reserved filtrate is for subsequent use;
5) step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, add antiseptic 0.01 ~ 0.02 weight portion, inactivator 0.2 ~ 0.3 weight portion, selenium 0.05 ~ 0.1 weight portion, antioxidant 1 ~ 2 weight portion, alpha-interferon 1 ~ 2 weight portion, active lactobacillus 0.5 ~ 1 weight portion, polyvinylpyrrolidone 1 ~ 2 weight portion, glycine 0.5 ~ 1 weight portion mix, first-20 DEG C of pre-coolings, lyophilizing under-80 DEG C of conditions, to obtain final product again.
7. the preparation method of the freeze-dried mixed powder of anti-chicken viral diseases according to claim 6, is characterized in that: step 2) described in the pH of acidified aqueous solution be 4.9 ~ 5.2; The addition of acidified aqueous solution is 8 times of yolk antibody liquor capacity.
8. the preparation method of the freeze-dried mixed powder of anti-chicken viral diseases according to claim 6, is characterized in that: sodium salt described in step 3) is NaCl and NaSO
4mass ratio be the mixture of 4:1.
9. the preparation method of the freeze-dried mixed powder of anti-chicken viral diseases according to claim 6, is characterized in that: the rotating speed of centrifugalize described in step 3) is 15000rpm, and the time is 20min.
10. the preparation method of the freeze-dried mixed powder of anti-chicken viral diseases according to claim 6, it is characterized in that: filtering described in step 4) is first filter with the filter membrane of 0.45 μm, molecular cut off is greater than the macromole of 200KD, then by the membrane filtration of filtrate with 0.22 μm.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105820242A (en) * | 2015-01-05 | 2016-08-03 | 郑州牧业工程高等专科学校 | Optimal extraction technology of infectious bursal diseased egg yolk immunoglobulin |
| CN106267194A (en) * | 2016-08-25 | 2017-01-04 | 青岛润达生物科技有限公司 | A kind of yolk antibody and Swine spleen transfer factor joint product |
| CN111480746A (en) * | 2020-04-26 | 2020-08-04 | 黑龙江省农业科学院畜牧兽医分院 | Preparation method and application of feed additive for resisting hydropericardium syndrome of chickens |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105820242A (en) * | 2015-01-05 | 2016-08-03 | 郑州牧业工程高等专科学校 | Optimal extraction technology of infectious bursal diseased egg yolk immunoglobulin |
| CN106267194A (en) * | 2016-08-25 | 2017-01-04 | 青岛润达生物科技有限公司 | A kind of yolk antibody and Swine spleen transfer factor joint product |
| CN111480746A (en) * | 2020-04-26 | 2020-08-04 | 黑龙江省农业科学院畜牧兽医分院 | Preparation method and application of feed additive for resisting hydropericardium syndrome of chickens |
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| CN104208675B (en) | 2016-08-17 |
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