Animal vaccination method

Abstract

The invention discloses an animal vaccine inoculation method, and relates to the technical field of animal vaccines. According to the inoculation method, an intelligent inoculation device is adopted for inoculation, the intelligent inoculation device comprises a vaccine loading inlet, a needleless injection mechanism, a synchronous marking mechanism, a control mechanism and a data management terminal, the vaccine loading inlet is communicated with the needleless injection mechanism, and the needleless injection mechanism comprises an injection head and a high-pressure metering pump. The synchronous marking mechanism comprises a marking block and a telescopic rod, the control mechanism is used for controlling the high-pressure metering pump and the telescopic rod, and the control mechanism is in Bluetooth connection with the data management terminal. Needle-free intradermal injection is adopted, rich antigen presenting cells in the corium layer are utilized, the immune response intensity is enhanced, and meanwhile the tedious operation caused by frequent needle replacement and the cross infection risk are avoided.

Description

Technical Field

[0001] The present invention relates to the technical field of animal vaccines, in particular to an animal vaccination method. Background Art

[0002] Vaccines are biological products made from various pathogenic microorganisms for use in vaccination. Vaccination is crucial to human health, and the production timelines for different vaccines vary. Vaccine development is a lengthy, complex, and costly process. Vaccination is the most economical and effective public health intervention for preventing and controlling infectious diseases. It is also an effective way for families to reduce illness and medical expenses.

[0003] Existing swine vaccination technologies present the following challenges: low immunization efficiency, requiring frequent needle changes and cumbersome procedures for traditional intramuscular injections; cross-infection risks, resulting from needle reuse or improper handling; and insufficient immune targeting, with traditional vaccines failing to target key immune escape factors and requiring limited adjuvant selection. Therefore, we propose an animal vaccination method to address these challenges.

[0004] The above information disclosed in this background technology is only for enhancing understanding of the background technology of the present invention and therefore it may contain information that does not constitute the prior art that is already known to a person of ordinary skill in the art. Summary of the Invention

[0005] The object of the present invention is to provide an animal vaccination method to solve the problems raised by the above background technology.

[0006] To achieve the above objectives, the present invention provides the following technical solutions: a method for animal vaccination, the method employing an intelligent vaccination device for vaccination, the intelligent vaccination device comprising a vaccine loading port, a needle-free injection mechanism, a synchronization marking mechanism, a control mechanism, and a data management terminal, the vaccine loading port being connected to the needle-free injection mechanism, the needle-free injection mechanism comprising an injection head and a high-pressure metering pump, the synchronization marking mechanism comprising a marking block and a telescopic rod, the control mechanism being configured to control the high-pressure metering pump and the telescopic rod, and the control mechanism being connected to the data management terminal via Bluetooth;

[0007] The inoculation method comprises the following steps:

[0008] Step 1: Fix the vaccine loading bottle at the vaccine loading inlet so that the vaccine loading bottle is connected to the high-pressure metering pump;

[0009] Step 2: Align the injection head with the site to be inoculated, and control the high-pressure metering pump through the control mechanism to form a jet flow of the metered drug through the injection head to penetrate the skin of the site to be inoculated at high speed to complete the intradermal injection;

[0010] Step 2.1: Primary immunization: 3-week-old piglets, intradermal injection of 0.2 ml;

[0011] Step 2.2: Booster immunization: repeat the same dose after 4 weeks;

[0012] Step 2.3: Pregnant sows, vaccinate 6 weeks before farrowing;

[0013] Step 3: The control mechanism controls the telescopic rod to push out the marking block, so that the marking block forms a mark on the surface of the pig skin;

[0014] Step 4: After vaccination, a biosensor is implanted subcutaneously to monitor body temperature and local inflammatory factors in real time, and the data is synchronized to the data management terminal;

[0015] Step 5: The data management terminal records the immunization time, dosage and individual marker information, generates an electronic immunization file, and combines it with the biosensor information for subsequent management.

[0016] Preferably, the injection head includes a positioning cylinder and an injection microchannel, and the injection microchannel runs through the central axis position of the injection head.

[0017] Preferably, the needle-free injection mechanism further includes a pressure sensor and an alarm module. The pressure sensor is used to monitor the injection pressure of the injection head in real time to ensure that the injection pressure is within the effective range. When the injection pressure deviates from the effective range, the alarm module is triggered to alarm.

[0018] Preferably, a fixing ring for fixing the vaccine loading bottle is provided above the vaccine loading inlet.

[0019] Preferably, the marking block includes a liquid storage bottle and liquid absorbent cotton, the liquid storage bottle is detachably mounted on the output end of the telescopic rod, the liquid absorbent cotton is mounted on the bottle mouth of the liquid storage bottle, and a support sheet is provided inside the liquid absorbent cotton.

[0020] Preferably, the surface of the intelligent vaccination device is provided with an anti-slip portion for hand-holding.

[0021] Preferably, the vaccine comprises SpA* antigen, LukABRARPR-33 antigen and dual adjuvants of AS01B and GLA-SE.

[0022] Preferably, the vaccine is prepared by:

[0023] Antigen preparation: SpA* and LukABRARPR-33 were recombinantly expressed in E. coli, purified by affinity chromatography, and mixed in a 1:1 ratio.

[0024] Adjuvant preparation: The adjuvant was encapsulated in poly(lactic-co-glycolic acid) (PLGA) microspheres;

[0025] Vaccine preparation: Mix the antigen and double adjuvant in a mass ratio of 5:1 to prepare a nanoemulsion with a particle size of ≤200nm.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The present invention adopts needle-free intradermal injection, utilizes the abundant antigen-presenting cells in the dermis, enhances the intensity of immune response, and avoids the tedious operation of frequent needle replacement and the risk of cross infection.

[0028] The above summary is for illustrative purposes only and is not intended to be limiting in any way. In addition to the illustrative aspects, embodiments and features described above, further aspects, embodiments and features of the present invention will be readily apparent by reference to the accompanying drawings and the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Schematic diagram of the inoculation method of the present invention;

[0030] Figure 2 This is a schematic diagram of the internal structure of the intelligent vaccination device of the present invention;

[0031] Figure 3 This is a schematic diagram of the main structure of the intelligent vaccination device of the present invention Figure 1 ;

[0032] Figure 4 This is a schematic diagram of the main structure of the intelligent vaccination device of the present invention Figure 2 . DETAILED DESCRIPTION

[0033] The following will provide a clear and complete description of the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0034] See also Figure 2 - Figure 4 A method for animal vaccination is disclosed, which uses an intelligent vaccination device 1 for vaccination. The intelligent vaccination device 1 includes a vaccine loading inlet 2, a needle-free injection mechanism 3, a synchronization marking mechanism 4, a control mechanism 5 and a data management terminal 6 (not shown in the figure), and the vaccine loading inlet 2 is connected to the needle-free injection mechanism 3.

[0035] The needle-free injection mechanism 3 includes an injection head 31 and a high-pressure metering pump 32. Specifically, the injection head 31 includes a positioning cylinder 311 and an injection microchannel 312. The injection microchannel 312 runs through the central axis of the injection head 31. The output end of the high-pressure metering pump 32 is connected to the injection microchannel 312.

[0036] Specifically, the inner wall of the vaccine loading inlet 2 is provided with a thread adapted to the vaccine loading bottle, a fixing ring 21 for fixing the vaccine loading bottle is provided above the vaccine loading inlet 2, and the bottom of the vaccine loading inlet 2 is connected to the high-pressure metering pump 32 through a connecting pipe.

[0037] Specifically, the needle-free injection mechanism 3 also includes a pressure sensor and an alarm module. The pressure sensor is used to monitor the injection pressure of the injection microchannel 312 in the injection head 31 in real time to ensure that the injection pressure is within the effective range (5KPa-8KPa). When the injection pressure deviates from the effective range, the alarm module is triggered to alarm.

[0038] The synchronous marking mechanism 4 comprises a marking block 41 and a telescopic rod 42. Specifically, the marking block 41 includes a liquid storage bottle 411 and a liquid absorbent sponge 412. The liquid storage bottle 411 is detachably mounted on the output end of the telescopic rod 42. The liquid absorbent sponge 412 is mounted on the mouth of the liquid storage bottle 411 and has a support sheet 413 disposed within the liquid absorbent sponge 412. Fluorescent dye is contained in the liquid storage bottle 411, and marking is performed by absorbing the fluorescent dye into the liquid absorbent sponge 412.

[0039] When the fluorescent dye is exhausted, the marking block 41 can be pushed out by the telescopic rod 42, and the marking block 41 can be removed and replaced. The fluorescent dye required for one marking is very small, so the marking block 41 does not need to be replaced frequently. Generally, it is sufficient to check whether the fluorescent dye is sufficient before use.

[0040] The control mechanism 5 is used to control the high-pressure metering pump 32 and the telescopic rod 42 . The control mechanism 5 is connected to the data management terminal 6 via Bluetooth.

[0041] Specifically, the surface of the intelligent vaccination device 1 is provided with an anti-slip portion 11 for hand-holding, which is convenient for staff to hold.

[0042] See also Figure 1 , the inoculation method comprises the following steps:

[0043] Step 1: Fix the vaccine loading bottle at the vaccine loading inlet 2 so that the vaccine loading bottle is connected to the high-pressure metering pump 32;

[0044] Step 2: Align the injection head 31 with the area to be inoculated, place the positioning cylinder 311 against the skin, keep the injection microchannel 312 relatively perpendicular to the area to be inoculated, and control the high-pressure metering pump 32 through the control mechanism 5 to inject the metered amount of drug through the injection head 31 to form a jet stream that penetrates the skin of the area to be inoculated at high speed, completing the intradermal injection;

[0045] Step 2.1: Primary immunization: 3-week-old piglets, intradermal injection of 0.2 ml;

[0046] Step 2.2: Booster immunization: repeat the same dose after 4 weeks;

[0047] Step 2.3: Pregnant sows, vaccinate 6 weeks before farrowing;

[0048] Step 3: The control mechanism 5 controls the telescopic rod 42 to push out the marking block 41, so that the absorbent cotton 412 on the marking block 41 forms a mark on the surface of the pig skin;

[0049] Step 4: After vaccination, a biosensor is implanted subcutaneously to monitor body temperature and local inflammatory factors in real time, and the data is synchronized to the data management terminal 6;

[0050] Step 5: The data management terminal 6 records the immunization time, dosage and individual marker information, generates an electronic immunization file, and combines it with the biosensor information for subsequent management.

[0051] The above-mentioned vaccine contains SpA* antigen, LukABRARPR-33 antigen and dual adjuvants of AS01B and GLA-SE. It uses genetically modified SpA* (detoxified staphylococcal protein A) and LukABRARPR-33 (modified leukocidin AB variant) to block the pathogen's immune escape; and uses a combination of AS01B (TLR4 agonist + saponin) and GLA-SE (TLR4 agonist emulsion) to activate TH1 and TH17 immune responses, respectively.

[0052] The preparation method of the above vaccine is as follows:

[0053] Antigen preparation: SpA* and LukABRARPR-33 were recombinantly expressed in E. coli, purified by affinity chromatography, and mixed in a 1:1 ratio.

[0054] Adjuvant preparation: The adjuvant was encapsulated in poly(lactic-co-glycolic acid) (PLGA) microspheres;

[0055] Vaccine preparation: Mix the antigen and double adjuvant in a mass ratio of 5:1 to prepare a nanoemulsion with a particle size of ≤200nm.

[0056] Vaccination speed test: Taking a fattening pig farm with 1,000 heads as an example, using 10 intelligent vaccination devices, a 3-person operation team can complete the immunization of the entire farm within 2 hours.

[0057] Virus challenge experiment: 40 healthy 3-week-old piglets weighing 8-10 kg were randomly divided into 4 groups, n = 10 per group; the vaccine group was vaccinated with a multi-component vaccine targeting Staphylococcus aureus SpA + LukAB; the adjuvant control group was vaccinated with only the adjuvant AS01B + GLA-SE at the same dose as the vaccine group; the positive control group was challenged but not vaccinated; the negative control group was not vaccinated and not challenged. The vaccine group and the adjuvant control group were inoculated using the vaccination method of the present invention. The highly pathogenic strain CC8 of Staphylococcus aureus was selected as the challenge pathogen, and the challenge dose was 1×10 8 CFU / head, using a surgical site infection model to simulate deep tissue infection. The challenge was performed 2 weeks after the booster immunization, and the animals were observed for 14 consecutive days after the challenge.

[0058] The detection indicators are as follows:

[0059] Clinical symptom scoring: The severity of dyspnea, fever (≥40°C), loss of appetite, and wound suppuration were divided into 0-4 levels and recorded daily.

[0060] Spleen bacterial load: Spleen tissue was collected on the 7th and 14th days after infection, homogenized, diluted and plated to determine CFU / g.

[0061] Inflammatory factor levels: ELISA was used to detect the concentrations of IL-6 and TNF-α in serum.

[0062] Antibody level: neutralizing antibody titer (ELISA method) and specific IgG / IgA titer.

[0063] Histopathological analysis: Splenic tissue sections were used to observe inflammatory cell infiltration and necrosis.

[0064] The test data of each group are as follows:

[0065] Detection indicators Vaccine group Adjuvant control group Positive control group Negative control group Clinical symptom score 1.2±0.3* 3.5±0.6 4.0±0.8 0±0 Splenic bacterial load <![CDATA[5×10 3 ±1.2×10 3 *]]> <![CDATA[8×10 4 ±2.1×10 4 ]]> <![CDATA[1×10 5 ±3.5×10 4 ]]> 0 Serum IL-6 (pg / mL) 120±25* 280±50 350±60 50±10 Neutralizing antibody titer (GMT) 1:6400* 1:800 <1:100 <1:100 Survival rate (day 14) 90％* 60％ 40％ 100％

[0066] Note: * indicates significant difference compared with the positive control group (P<0.05).

[0067] From the above, we can see that the average score of the vaccine group was 1.2, which was significantly lower than that of the positive control group, and the clinical symptoms were alleviated by 70%; the spleen bacterial load of the vaccine group decreased by 95%; the high and medium neutralizing antibody levels indicated that the vaccine induced a specific immune response; the inflammatory factor levels were reduced, indicating that the vaccine reduced tissue damage by inhibiting excessive inflammatory responses.

[0068] In the present invention, unless otherwise expressly specified or limited, the terms "mounted," "connected," "connect," "fixed," etc. should be understood broadly. For example, they may refer to fixed connection, detachable connection, or integration; mechanical connection or electrical connection; direct connection or indirect connection through an intermediate medium; internal communication between two components or interaction between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0069] In the present invention, unless otherwise expressly specified or limited, when a first feature is "above" or "below" a second feature, it may mean that the first and second features are in direct contact, or that the first and second features are in indirect contact through an intermediary. Furthermore, when a first feature is "above," "above," or "above" a second feature, it may mean that the first feature is directly above or diagonally above the second feature, or simply means that the first feature is at a higher level than the second feature. When a first feature is "below," "below," or "below" a second feature, it may mean that the first feature is directly below or diagonally below the second feature, or simply means that the first feature is at a lower level than the second feature.

[0070] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example" or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0071] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A method for animal vaccination, characterized in that: The vaccination method adopts an intelligent vaccination device (1) for vaccination, wherein the intelligent vaccination device (1) comprises a vaccine loading inlet (2), a needle-free injection mechanism (3), a synchronous marking mechanism (4), a control mechanism (5) and a data management terminal (6), wherein the vaccine loading inlet (2) is connected to the needle-free injection mechanism (3), the needle-free injection mechanism (3) comprises an injection head (31) and a high-pressure quantitative pump (32), the synchronous marking mechanism (4) comprises a marking block (41) and a telescopic rod (42), the control mechanism (5) is used to control the high-pressure quantitative pump (32) and the telescopic rod (42), and the control mechanism (5) is connected to the data management terminal (6) via Bluetooth; The inoculation method comprises the following steps: Step 1: Fix the vaccine loading bottle at the vaccine loading inlet (2) so that the vaccine loading bottle is connected to the high-pressure quantitative pump (32); Step 2: Align the injection head (31) with the site to be inoculated, and control the high-pressure metering pump (32) through the control mechanism (5) to inject the quantitative drug through the injection head (31) to form a jet flow that penetrates the skin of the site to be inoculated at high speed, thereby completing the intradermal injection; Step 2.1: Primary immunization: 3-week-old piglets, intradermal injection of 0.2 ml; Step 2.2: Booster immunization: repeat the same dose after 4 weeks; Step 2.3: Pregnant sows, vaccinate 6 weeks before farrowing; Step 3: Controlling the telescopic rod (42) through the control mechanism (5) to push out the marking block (41), so that the marking block (41) forms a mark on the surface of the pig skin; Step 4: After vaccination, a biosensor is implanted subcutaneously to monitor body temperature and local inflammatory factors in real time, and the data is synchronized to the data management terminal (6); Step 5: The data management terminal (6) records the immunization time, dosage and individual marker information, generates an electronic immunization file, and combines it with the biosensor information for subsequent management.

2. The animal vaccination method according to claim 1, wherein: The injection head (31) comprises a positioning cylinder (311) and an injection microchannel (312), and the injection microchannel (312) runs through the central axis position of the injection head (31).

3. The animal vaccination method according to claim 1, wherein: The needle-free injection mechanism (3) further comprises a pressure sensor and an alarm module. The pressure sensor is used to monitor the injection pressure of the injection head (31) in real time to ensure that the injection pressure is within an effective range. When the injection pressure deviates from the effective range, the alarm module is triggered to alarm.

4. The animal vaccination method according to claim 1, wherein: A fixing ring (21) for fixing the vaccine loading bottle is provided above the vaccine loading inlet (2).

5. The animal vaccination method according to claim 1, wherein: The marking block (41) comprises a liquid storage bottle (411) and a liquid absorbent cotton (412); the liquid storage bottle (411) is detachably mounted on the output end of the telescopic rod (42); the liquid absorbent cotton (412) is mounted on the bottle mouth of the liquid storage bottle (411); and a supporting sheet (413) is provided inside the liquid absorbent cotton (412).

6. The animal vaccination method according to claim 1, wherein: The surface of the intelligent vaccination device (1) is provided with an anti-slip portion (11) for hand-holding.

7. The animal vaccination method according to claim 1, wherein: The vaccine comprises SpA* antigen, LukABRARPR-33 antigen and dual adjuvants of AS01B and GLA-SE.

8. The animal vaccination method according to claim 7, characterized in that: The preparation method of the vaccine is: Antigen preparation: SpA* and LukABRARPR-33 were recombinantly expressed in E. coli, purified by affinity chromatography, and mixed in a 1:1 ratio. Adjuvant preparation: The adjuvant was encapsulated in poly(lactic-co-glycolic acid) (PLGA) microspheres; Vaccine preparation: Mix the antigen and double adjuvant in a mass ratio of 5:1 to prepare a nanoemulsion with a particle size of ≤200nm.

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