CN113017899B - Efficient large rabbit inoculation method - Google Patents

Efficient large rabbit inoculation method Download PDF

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CN113017899B
CN113017899B CN202110253834.3A CN202110253834A CN113017899B CN 113017899 B CN113017899 B CN 113017899B CN 202110253834 A CN202110253834 A CN 202110253834A CN 113017899 B CN113017899 B CN 113017899B
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inoculation
big
rabbits
rabbit
seed virus
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CN113017899A (en
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张惠云
江兴华
林志坤
陈淑亚
蔡杰
包松英
陈慧婧
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Beijing Biomedical Technology Center Of Zhaofenghua Biotechnology Nanjing Co ltd
Beijing Kemufeng Biological Pharmaceutical Co ltd
Zhaofenghua Biotechnology Nanjing Co ltd
Zhaofenghua Biotechnology Fuzhou Co ltd
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Beijing Biomedical Technology Center Of Zhaofenghua Biotechnology Nanjing Co ltd
Beijing Kemufeng Biological Pharmaceutical Co ltd
Zhaofenghua Biotechnology Nanjing Co ltd
Zhaofenghua Biotechnology Fuzhou Co ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61DVETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
    • A61D1/00Surgical instruments for veterinary use
    • A61D1/02Trocars or cannulas for teats; Vaccination appliances
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61DVETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
    • A61D3/00Appliances for supporting or fettering animals for operative purposes
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61DVETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
    • A61D7/00Devices or methods for introducing solid, liquid, or gaseous remedies or other materials into or onto the bodies of animals

Abstract

The invention discloses an efficient big rabbit inoculation method, which uses a conveyer belt and a seed virus heat preservation device, and comprises the following specific steps: 1) placing the big rabbits in a retaining cage, and conveying the big rabbits to an inoculation table for inoculation through a conveying belt; 2) inoculating the big rabbits by a continuous injector, wherein in the inoculation process of the big rabbits, a conveying belt keeps static, and a magnetic stirrer works and is used for continuously and uniformly mixing the seed viruses in a seed virus bottle; 3) after the big rabbits are inoculated, hemostasis gel is adopted to stop bleeding of the inoculation position of the big rabbits, after inoculation and hemostasis operation of one big rabbit is completed, the conveying belt continues to convey forwards to send the big rabbits in the subsequent retaining cage into the inoculation table, the inoculation and hemostasis operation is repeated, and finally the big rabbits after inoculation and hemostasis are sequentially sent back to the rabbit cage through the conveying belt. The invention can reduce the walking of personnel in the inoculation process and reduce the pollution risk; the heat preservation container is matched with the magnetic stirrer to be used for heat preservation and uniform mixing of the seed virus, so that the loss of seed virus titer in the inoculation link is reduced to the minimum.

Description

Efficient large rabbit inoculation method
Technical Field
The invention relates to the technical field of big rabbit inoculation, in particular to a high-efficiency big rabbit inoculation method.
Background
Preparing 20-50 times diluted emulsion of the seed venom by using sterilized normal saline, and placing the diluted seed venom in a stainless steel basin containing glacial alum for refrigeration and heat preservation. The seed virus dilutions were injected with a 5ml syringe, 1ml per ear vein of rabbits, and the rabbits were grabbed from the cages by the manufacturer.
The preparation process of the swine fever live vaccine (rabbit source) antigen has the following 3 problems:
1. the big rabbits are held by hands for fixing in the inoculation process of the big rabbits, so that the big rabbits are easy to panic and stress, and fixing personnel also have the risk of being scratched by the big rabbits;
2. in the traditional method, a common injector is used for inoculating the rabbits, the time consumption is long, the efficiency is low, the seed viruses are placed for a long time at normal temperature during inoculation, the hog cholera viruses are easy to die, the titer of the seed viruses is reduced, and the reaction rate of the inoculated rabbits is influenced, so that the product quality is influenced;
3. since big rabbit ear edge intravenous route vaccine poison, press the hemostasis after the injection, this method is not only consuming time, and still can continue to bleed after pressing, leads to the seed poison part to run off along with bleeding, influences the virus and breeds in the rabbit body, reduces the finished product titer, increases the cost of product, and on the other hand still can bite the wound easily with the cage rabbit, causes the wound infection.
Disclosure of Invention
The invention aims to provide an efficient rabbit inoculation method.
In order to achieve the purpose, the invention adopts the following technical scheme:
an efficient rabbit inoculation method comprises a conveying belt and a seed virus heat preservation device, wherein the conveying belt is annular and horizontally arranged, and a plurality of retaining cages are arranged on the conveying belt at intervals along the conveying direction;
the seed poison heat preservation device comprises a container and a magnetic stirrer, wherein the container is arranged on the magnetic stirrer, a cooling interlayer is arranged on the side wall of the container, and a refrigerant inlet is formed in the top of the cooling interlayer; before inoculation, a seed virus bottle containing seed virus is placed in a container, a refrigerant is put into a cooling interlayer of the container, magnetic particles are placed in the seed virus bottle, a pipeline at the inlet of a continuous injector enters the seed virus bottle through a plug at the bottle mouth of the seed virus bottle, air in the continuous injector is discharged, and after the injection amount is adjusted to be 1 ml/time, rabbit inoculation can be carried out;
the method for inoculating the big rabbits comprises the following specific steps:
1) placing the big rabbits in a holding cage, and conveying the big rabbits to an inoculation table for inoculation through a conveying belt;
2) inoculating the big rabbits by a continuous injector, wherein in the inoculation process of the big rabbits, a conveying belt keeps static, and a magnetic stirrer works and is used for continuously and uniformly mixing the seed viruses in a seed virus bottle;
3) after the big rabbits are inoculated, hemostasis gel is adopted to stop bleeding of the inoculation position of the big rabbits, after inoculation and hemostasis operation of one big rabbit is completed, the conveying belt continues to convey forwards to send the big rabbits in the subsequent retaining cage into the inoculation table, the inoculation and hemostasis operation is repeated, and finally the big rabbits after inoculation and hemostasis are sequentially sent back to the rabbit cage through the conveying belt.
During the inoculation process of the big rabbits, the relaxing music is played in the inoculation table.
The preparation method of the hemostatic gel comprises the following steps: uniformly stirring gelatin and water to obtain a gelatin aqueous solution, adding streptomycin and D-glucosamine into the gelatin aqueous solution, and uniformly mixing to obtain the hemostatic gel, wherein the gelatin accounts for 10-20% of the hemostatic gel by mass, the streptomycin accounts for 1% of the hemostatic gel by mass, and the concentration of the D-glucosamine in the hemostatic gel is 2-4 mmol/L; then filling the hemostatic gel into a 20ml syringe, placing the syringe in a constant temperature water bath kettle at 40 ℃, and when in use, extruding the hemostatic gel into the inoculation part of the big rabbit, and achieving the purpose of hemostasis after the hemostatic gel is solidified.
By adopting the technical scheme, the invention has the following beneficial technical effects:
1. the horizontal conveying belt is arranged, so that the big rabbits are fixed on the conveying belt, the inoculation link operation is performed, the walking of personnel in the inoculation process is reduced, the pollution risk is reduced, the risk of overlarge rabbit stress caused by long-time rabbit grasping of the personnel is reduced, and the risk of scratching of the personnel is reduced;
2. a heat preservation container is adopted to be matched with a magnetic stirrer for heat preservation and uniform mixing of the seed virus, and meanwhile, a continuous injector is connected in the container for inoculation of the big rabbits, so that the loss of seed virus titer in an inoculation link is reduced to the minimum;
3. the self-made hemostatic gel is used for hemostasis of the inoculated part of the big rabbit, the hemostatic effect is good, the consumed time is short, the streptomycin can effectively inhibit bacteria, and simultaneously the D-glucosamine can promote virus proliferation.
Drawings
The invention is described in further detail below with reference to the accompanying drawings and the detailed description;
FIG. 1 is a schematic view of a general conveyor belt of the present invention;
FIG. 2 is a schematic view of the incubation device for seed virus of the present invention.
Detailed Description
As shown in figure 1 or figure 2, the high-efficiency rabbit inoculation method disclosed by the invention uses a conveyer belt 1 and a seed virus heat preservation device 6, wherein the conveyer belt 1 is annular and horizontally arranged, and a plurality of retaining cages 2 are arranged on the conveyer belt 1 at intervals along the conveying direction;
the seed poison heat preservation device 6 comprises a container 61 and a magnetic stirrer 62, wherein the container 61 is arranged on the magnetic stirrer 62, a cooling interlayer 611 is arranged on the side wall of the container 61, and a refrigerant inlet is formed in the top of the cooling interlayer 611; before inoculation, a seed virus bottle 63 filled with seed viruses is placed in a container 61, a cooling medium (ice alum) is put into a cooling interlayer 611 of the container 61, magnetic particles 64 are placed in the seed virus bottle 63, a pipeline 64 at the inlet of a continuous injector enters the seed virus bottle 63 through a plug at the bottle mouth of the seed virus bottle 63, air in the continuous injector is discharged, and after the injection amount is adjusted to be 1 ml/time, the inoculation of the big rabbit can be carried out.
Before inoculation, selecting and feeding the raw material big rabbits:
the normal body temperature of the big rabbit is 38.2-39.5 ℃, and the big rabbit with good nutrition state and the weight of 1.5-3kg is selected as the animal for preparing the vaccine. Before inoculation, the temperature of the big rabbits should be measured and observed for at least 3 days, and the temperature is measured once in the morning and afternoon every day. Selecting big rabbits with normal body temperature and small temperature difference fluctuation;
and seed toxin preparation: selecting qualified hog cholera lapinized virus strain (C strain) seed virus, and grinding and diluting the seed virus with a certain dilution to obtain the seed virus for production.
The inoculation method of the big rabbit comprises the following specific steps:
1) placing the big rabbits into a holding cage 2, and conveying the big rabbits to an inoculation table for inoculation through a conveying belt 1;
2) inoculating the big rabbits by a continuous injector, wherein in the inoculation process of the big rabbits, the conveyer belt 1 keeps static, and the magnetic stirrer 62 works and is used for continuously and uniformly mixing the seed viruses in the seed virus bottle 63; in the inoculation process of the big rabbits, the relaxing music is played in the inoculation table, so that the big rabbits calm down, and the stress is reduced;
3) adopt hemostasis gel to stanch to the inoculation position (ear edge vein) of big rabbit after the big rabbit inoculation, accomplish the inoculation of a big rabbit and stanch the operation after, during conveyer belt 1 continued forward transport sent the big rabbit in the continuous cage 2 of keeping into the inoculation platform, repeated above-mentioned inoculation and hemostasis operation, the big rabbit after will inoculating and stanching is sent back the rabbit-hutch in proper order by conveyer belt 1 at last, wherein opening of conveyer belt 1 stops realizing by manual operation button.
The preparation method of the hemostatic gel adopted in the invention comprises the following steps: uniformly stirring gelatin and water to obtain a gelatin aqueous solution, adding streptomycin and D-glucosamine into the gelatin aqueous solution, and uniformly mixing to obtain the hemostatic gel, wherein the gelatin accounts for 10-20% of the hemostatic gel by mass, the streptomycin accounts for 1% of the hemostatic gel by mass, and the concentration of the D-glucosamine in the hemostatic gel is 2-4 mmol/L; then filling the hemostatic gel into a 20ml syringe, placing the syringe in a constant temperature water bath kettle at 40 ℃, and when in use, extruding the hemostatic gel into the inoculation part of the big rabbit, and achieving the purpose of hemostasis after the hemostatic gel is solidified.
After the inoculation of the big rabbits, the following steps are required:
and (3) harvesting the antigen:
after the inoculation of the big rabbits, the body temperature was measured once in the morning and afternoon, and 24 hours later, the body temperature was measured once every 6 hours. And selecting the setting heat reaction rabbit for harvesting. The criteria for styling heat-reacted rabbits are as follows: the incubation period is 24-48 hours, the body temperature rise is obvious curve, the temperature is 1 ℃ above the normal temperature, at least 3 times, and the incubation time is 18-36 hours.
Selecting the rabbit with the stereotyped heat reaction, dissecting and killing the rabbit from the temperature reduction and within 24 hours later, and adopting splenomegaly and lymph nodes by aseptic operation. When collecting poison, the stomach, intestine and esophagus are not cut and cut to avoid pollution.
And (3) subsequent processes of freeze-drying the matched seedlings and the like:
the harvested spleen and lymph nodes of the big rabbit are removed of adipose tissues under aseptic operation, and the prepared tissue is weighed and crushed. Adding 5% sucrose skimmed milk protectant, grinding, and filtering. The residue is deducted, and then calculated according to the actual filtered tissue quantity, and a proper amount of milk sucrose protective agent is added according to the requirement. Fully and uniformly stirred, subpackaged and freeze-dried.
And (4) performing titer inspection:
after the freeze-drying is finished, detection is carried out according to the requirements of the veterinary biological product regulation of the people's republic of China, and the average value of the titer of the finished product of the swine fever spleen gonorrhea vaccine attenuated vaccine prepared according to the method is 900 RID/head.
The vaccine produced by the new process and the traditional process is subjected to finished product titer detection, and the result is shown in the following table:
Figure BDA0002959363460000041
note "+ +" indicates pass and "-" indicates fail.
From the above table, it can be seen that the total of 9 batches of 750 RIDs in 10 batches of vaccines by the conventional process has 1 batch of 750 RIDs, 1 batch of 800 RIDs, 1 batch of 850 RIDs and 8 batches of 900 RIDs in the process of the present invention, and the titer of the vaccine prepared by the novel method is higher than that of the vaccine produced by the conventional process.
While the invention has been described in connection with the above embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, which are illustrative and not restrictive, and that those skilled in the art will understand that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the present invention, and they should be construed as being included in the following claims and description.

Claims (2)

1. An efficient rabbit inoculation method, which is used for a conveyer belt and a seed virus heat preservation device, is characterized in that:
the conveying belt is annular and horizontally arranged, and a plurality of retaining cages are arranged on the conveying belt at intervals along the conveying direction of the conveying belt;
the seed poison heat preservation device comprises a container and a magnetic stirrer, wherein the container is arranged on the magnetic stirrer, a cooling interlayer is arranged on the side wall of the container, and a refrigerant inlet is formed in the top of the cooling interlayer; before inoculation, a seed virus bottle containing seed virus is placed in a container, a refrigerant is put into a cooling interlayer of the container, magnetic particles are placed in the seed virus bottle, a pipeline at the inlet of a continuous injector enters the seed virus bottle through a plug at the bottle mouth of the seed virus bottle, air in the continuous injector is removed, and after the injection amount is adjusted to be 1 ml/time, the inoculation of the big rabbit can be carried out;
the method for inoculating the big rabbits comprises the following specific steps:
1) placing the big rabbits in a holding cage, and conveying the big rabbits to an inoculation table for inoculation through a conveying belt;
2) inoculating the big rabbits by a continuous injector, wherein in the inoculation process of the big rabbits, a conveying belt keeps static, and a magnetic stirrer works and is used for continuously and uniformly mixing the seed viruses in a seed virus bottle;
3) after the big rabbits are inoculated, hemostasis gel is adopted to stop bleeding of the inoculation position of the big rabbits, after inoculation and hemostasis operation of one big rabbit is completed, the conveyer belt is continuously conveyed forwards to send the big rabbits in the subsequent retaining cage into the inoculation table, the inoculation and hemostasis operation is repeated, and finally the big rabbits after inoculation and hemostasis are sequentially sent back to the rabbit cage by the conveyer belt;
the preparation method of the hemostatic gel comprises the following steps: uniformly stirring gelatin and water to obtain a gelatin aqueous solution, adding streptomycin and D-glucosamine into the gelatin aqueous solution, and uniformly mixing to obtain the hemostatic gel, wherein the gelatin accounts for 10-20% of the hemostatic gel by mass, the streptomycin accounts for 1% of the hemostatic gel by mass, and the concentration of the D-glucosamine in the hemostatic gel is 2-4 mmol/L; then filling the hemostatic gel into a 20ml syringe, placing the syringe in a constant temperature water bath kettle at 40 ℃, and when in use, extruding the hemostatic gel into the inoculation part of the big rabbit, and achieving the purpose of hemostasis after the hemostatic gel is solidified.
2. A high efficiency method for inoculating large rabbits according to claim 1, wherein: during the inoculation process of the big rabbits, the relaxing music is played in the inoculation table.
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CN107411843A (en) * 2017-05-15 2017-12-01 青岛澳德思瑞智能免疫技术有限公司 A kind of immune continuous syringe of intelligent automatic fowl

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Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN204581583U (en) * 2015-03-10 2015-08-26 福建省农业科学院畜牧兽医研究所 Rabbit vaccinate movable operating platform device
CN205019528U (en) * 2015-09-18 2016-02-10 马卫平 Closed bacterin continuous syringe
CN107411843A (en) * 2017-05-15 2017-12-01 青岛澳德思瑞智能免疫技术有限公司 A kind of immune continuous syringe of intelligent automatic fowl

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