CN113893339A - Preparation method of triple inactivated vaccine for preventing forest musk deer pneumonia - Google Patents
Preparation method of triple inactivated vaccine for preventing forest musk deer pneumonia Download PDFInfo
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Abstract
The preparation method of the triple inactivated vaccine for preventing the pneumonia of the forest musk deer comprises the following steps of: step 1, separating pathogenic escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae from pathogenic forest musk deer, and preserving the strains by adopting a freeze drying method; step 2, respectively placing the three bacteria in a culture medium for culture at 37 ℃, performing amplification culture, and inactivating the bacteria liquid for 24 hours by using formaldehyde with the concentration of 4 per mill; step 3, diluting each strain to 8.0 × 10 with sterilized normal saline9cfu/mL; step 4, mixing pathogenic escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae according to the ratio of 2: 2: mixing the components according to the proportion of 1, adding 1 ten thousand units of streptomycin, and uniformly mixing the components by a stirring device to obtain the triple inactivated vaccine. The vaccine prepared by the method has low price, easy operation and good immune effect.
Description
Technical Field
The invention belongs to the field of animal husbandry and veterinary medicine, and particularly relates to a preparation method of a triple inactivated vaccine for preventing woodmusk deer pneumonia.
Background
The pneumonia of forest musk deer is an acute or chronic respiratory disease of forest musk deer caused by various bacteria, and is characterized by runny nose, dyspnea and lung inflammation injury. Along with the multiplication of the breeding quantity of the forest musk deer, the pneumonia prevalence of the forest musk deer is in an increasing trend, and the death rate of the forest musk deer caused by pneumonia can be higher than 8% of the total number of the forest musk deer in the stock of the forest musk deer every year, so that the forest musk deer is developed to be the most important restriction factor for breeding the forest musk deer and protecting the forest musk deer.
According to the research on the causes of a large number of died cases of the forest musk deer pneumonia, the Escherichia coli and the Klebsiella pseudoaeruginosa nucleopneumoniae are the most frequently separated bacterial species in the died cases of the forest musk deer pneumonia, so that the 3 bacteria are selected as vaccine research objects to develop the forest musk deer pneumonia vaccine.
Escherichia coli is an important conditional pathogen, widely exists in the environment and animal bodies, and is an important pathogen of various animals and human beings. Escherichia coli can cause pneumonia of the forest musk deer, and causes disease symptoms such as lung hemorrhage, lung excess change and the like. The Escherichia coli comprises four antigens of an O antigen, a K antigen, an H antigen and an F antigen. The O antigen is lipopolysaccharide, has 171 kinds, and is the basis of grouping
Pseudomonas aeruginosa is an important conditional pathogen, widely exists in the environment, and is an important pathogen of various animals and people. Can cause acute hemorrhagic pneumonia or chronic pulmonary abscess of the forest musk deer. The pseudomonas aeruginosa comprises O antigen, H antigen, mucus antigen, R antigen, pilus antigen and the like. The bacteria can be divided into 20 serotypes according to O antigen serological typing.
Klebsiella pneumoniae Enterobacteriaceae, a gram-negative crude Brevibacterium, is an important conditional pathogen. The resistance to the outside is strong, and the drug resistance is easy to generate for most antibiotics. Like other bacteria of the enterobacteriaceae family, have O antigens and K antigens (i.e., somatic and capsular antigens).
The preparation process of the inactivated vaccine is widely applied in the world, and the vaccine has good immune effect and good safety and is the first choice for preventing animal epidemic diseases. The inactivated vaccine is a biological product which is mainly used for generating specific immunoprotection for animals by subcutaneous or intramuscular injection after pathogen is inactivated by formaldehyde. The traditional Chinese medicine composition is low in price, easy to operate and good in immune effect, and is widely applied to veterinary clinic.
Disclosure of Invention
Therefore, the invention provides a preparation method of a triple inactivated vaccine for preventing forest musk pneumonia, which comprises the following steps:
step 1, separating pathogenic escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae from pathogenic forest musk deer, and preserving the strains by adopting a freeze drying method;
step 2, respectively placing the three bacteria in a culture medium for culture at 37 ℃, performing amplification culture, and inactivating the bacteria liquid for 24 hours by using formaldehyde with the concentration of 4 per mill;
Step 4, mixing pathogenic escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae according to the ratio of 2: 2: mixing the components in a ratio of 1, adding 1 ten thousand units of streptomycin, and uniformly mixing the components by a stirring device to obtain the triple fire-extinguishing vaccine.
Preferably, the stirring device includes: a transmission case, a first motor, a first rotating shaft, a crank, a first gear, a deflector rod, a guide rail, a slide block and a rack,
the bottom end of the transmission box is provided with an opening, the bottom wall of the transmission box is provided with a first motor, an output shaft of the first motor is coaxially connected with one end of a first rotating shaft, the other end of the first rotating shaft is connected with one end of a crank, a first gear is coaxially arranged on the first rotating shaft, the other end of the crank is vertically connected with one end of a deflector rod, the top of the inner wall of the transmission box is provided with a guide rail in the horizontal direction, the guide rail is connected with a sliding block in a sliding manner, the bottom end of the sliding block is provided with a rack, one end of a first L-shaped connecting rod is connected with one end of the sliding block, the other end of the first L-shaped connecting rod is provided with a sliding groove extending along the length direction, and the other end of the deflector rod penetrates through the sliding groove;
a second gear is arranged on one side of the rack, the rack is meshed with the second gear, a second rotating shaft is coaxially arranged on the second gear, one end of the second rotating shaft is connected with the second gear, the other end of the second rotating shaft penetrates through a first fixing block to be connected with the middle of a rotating rod in the horizontal direction, the second rotating shaft is rotatably connected with the first fixing block, the first fixing block is connected with the inner wall of the transmission box, two ends of the rotating rod are respectively provided with a first connecting rod in the vertical direction, the lower end of the first connecting rod is coaxially connected with a third gear, a ring is arranged on one side of the first gear, both the inner ring and the outer ring of the ring are provided with teeth, the outer ring of the ring is meshed with the first gear, the inner ring of the ring is respectively meshed with the two third gears, a sleeve is sleeved on the second rotating shaft, and the sleeve is rotatably connected with the second rotating shaft, the telescopic left and right sides is provided with second L shape pole respectively, second L shape pole one end with telescopic outer wall connection, the second L shape pole other end with the ring upper end is connected, two be provided with the fourth gear between the third gear, the fourth gear with the third gear meshing, transmission case bottom is provided with the barrel, the third gear bottom is provided with the third pivot of vertical direction, third pivot one end with third gear coaxial coupling, the third pivot other end passes the opening is connected with first stirring leaf, the fourth gear with be provided with the second fixed block between the opening, the second fixed block with transmission case inner wall connection, fourth gear bottom is provided with the fourth pivot of vertical direction, fourth pivot one end with fourth gear coaxial coupling, the fourth pivot other end passes in proper order the second fixed block, The opening is connected with the second stirring blade, the fourth rotating shaft is connected with the second fixing block in a rotating mode, a liquid inlet pipe is arranged on one side of the barrel, and a liquid outlet pipe is arranged on the other side of the barrel.
Preferably, a second connecting rod in the vertical direction is arranged at the bottom end of the circular ring, the lower end of the second connecting rod penetrates through the opening to be connected with the sweeper, and bristles of the sweeper are in contact with the inner wall of the cylinder.
Preferably, the liquid inlet pipe and the liquid outlet pipe are both provided with electromagnetic valves.
Preferably, the side wall of the transmission case is provided with an access hole, and the access hole is provided with a sealing plate.
Preferably, the sealing plate is connected with the transmission case through a screw.
Preferably, a dust filter screen is arranged on the outer wall of the transmission case.
Preferably, the bottom end of the cylinder body is provided with a universal wheel.
The technical scheme of the invention has the following advantages: the preparation method has biological safety, and the vaccine strain is selected and fully inactivated by formaldehyde; the vaccine has the function of preventing the forest musk pneumonia caused by escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae, all vaccine strains of the vaccine are separated from Shanxi province, and are determined to be epidemic dominant strains through molecular typing identification; the invention has the characteristics of strong immunogenicity and good specificity, and the invention can inactivate all bacteria and generate strong immunity; the separated strain is obtained from screening of a large number of clinical cases, has better coverage, and has the characteristics of simple preparation method and low cost.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and drawings.
The technical solution of the present invention is further described in detail by the accompanying drawings and embodiments.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 is a schematic view of the structure of a stirring apparatus according to the present invention;
FIG. 2 is a schematic view of the first L-shaped connecting rod and the shift lever of the present invention;
FIG. 3 is a schematic view of the engagement of the ring, the first gear, the third gear and the fourth gear in the present invention;
wherein, 1-a transmission box, 2-an opening, 3-a first motor, 4-a first rotating shaft, 5-a crank, 6-a first gear, 7-a deflector rod, 8-a guide rail, 9-a slide block, 10-a rack, 11-a first L-shaped connecting rod, 12-a chute, 13-a second gear, 14-a second rotating shaft, 15-a first fixed block, 16-a rotating rod, 17-a first connecting rod, 18-a third gear, 19-a circular ring, 20-a sleeve, 21-a second L-shaped rod, 22-a fourth gear, 23-a cylinder, 24-a third rotating shaft, 25-a first stirring blade, 26-a second fixed block, 27-a fourth rotating shaft, 28-a second stirring blade, 29-a liquid inlet pipe and 30-a liquid outlet pipe, 31-a second connecting rod, 32-a sweeping brush, 33-an electromagnetic valve, 34-an access hole, 35-a sealing plate, 36-a dust filtering net and 37-a universal wheel.
Detailed Description
In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more clearly apparent, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or be indirectly on the other element. When an element is referred to as being "connected to" another element, it can be directly or indirectly connected to the other element.
It will be understood that the terms "length," "width," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like, as used herein, refer to an orientation or positional relationship indicated in the drawings that is solely for the purpose of facilitating the description and simplifying the description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and is therefore not to be construed as limiting the invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
Example one
The invention provides a preparation method of a triple inactivated vaccine for preventing forest musk pneumonia, which comprises the following steps:
1. strain rejuvenation
Inoculating the three strains of bacteria to a mouse at a dose of 10LD50 respectively, killing the mouse after the mouse is attacked, opening the chest cavity under aseptic conditions, cutting the lung, dipping and streaking the bacteria by using an inoculating ring, inoculating the bacteria to an LB solid culture medium, culturing the bacteria at 37 ℃ for 16h, selecting a single colony, inoculating 5mL of the LB liquid culture medium, performing shake culture at 220rpm and 37 ℃, preserving the strains, and performing amplification culture according to a ratio of 1: 1000.
2. Preparation of inactivated vaccine
(1) Preparation and examination of bacterial liquid
Preparing LB culture medium according to the demand, inoculating 3 kinds of bacteria according to the proportion of 1:1000, culturing at 37 deg.C for 14-18h, and counting viable bacteria.
(2) Inactivation and concentration of bacterial liquid
Adding formaldehyde solution into the culture solution at a ratio of 1:1000, acting at 37 deg.C and 120rpm for 48h, centrifuging at 12000rpm to obtain inactivated thallus, washing with normal saline for 3 times, and adjusting thallus concentration to 8.0 × 109cfu/mL, sterile test.
(3) Preparation of trivalent vaccine
The three monovalent inactivated bacteria with the adjusted concentration are mixed according to the ratio of 2: 2: 1, adding streptomycin with the final concentration of 1 ten thousand units, uniformly mixing by a stirring device to obtain the triple fire-extinguishing vaccine, and respectively preparing 3 batches of vaccines, wherein the vaccines are numbered 202101, 202102 and 202103.
(4) Vaccine dispensing
And after the sterile inspection is qualified, subpackaging according to 100Ml per bottle, sealing and storing at 2-8 ℃.
Example two
1. Security check
The test items (sterility test, safety test, test for residual formaldehyde content and physical property test) were carried out according to the "protocol for veterinary biological product manufacture and test" (hereinafter referred to as "protocol").
1.1 sterility test
The operation is carried out according to the regulation, and the three batches of vaccines are qualified through sterile inspection, 202101, 202102 and 202103.
1.2 physical Property test
Observing color, shape, etc. under static and shaking state respectively; shaking to obtain uniform milky white suspension; standing, wherein the upper layer is clear liquid, and the lower layer is white precipitate;
1.3 examination of residual Formaldehyde quantity
Operating according to the regulations; the detected residual amount of formaldehyde is less than 0.10 percent and is in the allowable range of the regulation.
1.4 overdose inoculation safety test
5 rabbits were immunized with 5ml of inactivated vaccine, a normal saline control group was established, and the feeding, mental state, and local absorption of the vaccinated animals were observed within 30 days. In the injection vaccine 30d, the rabbit has no obvious difference in food intake and mental state from the control, and has good local absorption. The results show that: the product has good biological safety.
2. Efficacy test
2.1 Linshe triple pneumonia vaccine for mouse immune challenge protection experiment
40 mice were divided into 4 groups of 10 mice each. 3, 3 and 4 mice in the 1 st group and the 2 nd group are respectively used for immunizing 202101, 202102 and 202103, and the batch of the triple vaccine is 0.2mL, and a second needle is inoculated after 14 days; group 3 and group 4 were inoculated with 0.2mL of physiological saline, respectively.
Inoculating 10LD50 Escherichia coli in groups 1 and 3 after the second immunization at 14 d; groups 2 and 4 were inoculated with 10LD50 Pseudomonas aeruginosa. Observed at 14d after inoculation, the result shows that all mice in the immunization group survive, and 1 survived in the 4 th group of the mice in the control group. The vaccine immunization can provide good immune protection for the mice, and the table 1 shows.
TABLE 1 protection test results of Linshe pneumonia triple vaccine on mice
2.2 Linshe triple vaccine immunization Linshe induced antibody response
In three fields, 2 mL/young forest musk deer born in the current year is immunized, blood is collected at 30d, 44d and 60d after first immunization, and antibodies of escherichia coli and pseudomonas aeruginosa are detected, and the antibodies are shown in tables 2 and 3.
Physical properties and preservation of vaccines
(1) The product is milky white suspension; standing, wherein the upper layer is clear liquid, and the lower layer is white precipitate;
(2) the effective period is 12 months at 2-8 ℃ from the day of the self-made vaccine;
(3) the vaccine is applied to forest musk deer
Note that: firstly, only intramuscular injection is used for emergency inoculation in an epidemic area; secondly, a small group is firstly subjected to a safety test, and the safety test proves that the safety test can be used in a large area; thirdly, the product is ineffective in drinking water and mixing materials; and fourthly, if serious reaction of vaccination occurs, 0.1 percent of adrenaline can be used for emergency rescue.
EXAMPLE III
As shown in fig. 1 to 3, the stirring apparatus includes: a transmission case 1, a first motor 3, a first rotating shaft 4, a crank 5, a first gear 6, a deflector rod 7, a guide rail 8, a slide block 9 and a rack 10,
the bottom end of the transmission case 1 is provided with an opening 2, the bottom wall of the transmission case 1 is provided with a first motor 3, an output shaft of the first motor 3 is coaxially connected with one end of a first rotating shaft 4, the other end of the first rotating shaft 4 is connected with one end of a crank 5, a first gear 6 is coaxially arranged on the first rotating shaft 4, the other end of the crank 5 is vertically connected with one end of a deflector rod 7, the top of the inner wall of the transmission case 1 is provided with a guide rail 8 in the horizontal direction, the guide rail 8 is slidably connected with a slide block 9, the bottom end of the slide block 9 is provided with a rack 10, one end of a first L-shaped connecting rod 11 is connected with one end of the slide block 9, the other end of the first L-shaped connecting rod 11 is provided with a chute 12 extending along the length direction, and the other end of the deflector rod 7 passes through the chute 12;
a second gear 13 is arranged on one side of the rack 10, the rack 10 is engaged with the second gear 13, a second rotating shaft 14 is coaxially arranged on the second gear 13, one end of the second rotating shaft 14 is connected with the second gear 13, the other end of the second rotating shaft 14 passes through a first fixed block 15 and is connected with the middle of a rotating rod 16 in the horizontal direction, the second rotating shaft 14 is rotatably connected with the first fixed block 15, the first fixed block 15 is connected with the inner wall of the transmission case 1, two ends of the rotating rod 16 are respectively provided with a first connecting rod 17 in the vertical direction, the lower end of the first connecting rod 17 is coaxially connected with a third gear 18, one side of the first gear 6 is provided with a ring 19, the inner rings of the ring 19 and the outer rings of the ring 19 are both provided with teeth, the outer ring of the ring 19 is engaged with the first gear 6, the inner rings of the ring 19 are respectively engaged with the two third gears 18, the sleeve 20 is sleeved on the second rotating shaft 14, the sleeve 20 is rotatably connected with the second rotating shaft 14, the left side and the right side of the sleeve 20 are respectively provided with a second L-shaped rod 21, one end of the second L-shaped rod 21 is connected with the outer wall of the sleeve 20, the other end of the second L-shaped rod 21 is connected with the upper end of the circular ring 19, a fourth gear 22 is arranged between the two third gears 18, the fourth gear 22 is meshed with the third gears 18, the bottom end of the transmission case 1 is provided with a cylinder 23, the bottom end of the third gears 18 is provided with a third rotating shaft 24 in the vertical direction, one end of the third rotating shaft 24 is coaxially connected with the third gears 18, the other end of the third rotating shaft 24 penetrates through the opening 2 to be connected with the first stirring blade 25, the fourth gear 22 is provided with a second fixing block 26 between the opening 2, and the second fixing block 26 is connected with the inner wall of the transmission case 1, fourth gear 22 bottom is provided with the fourth pivot 27 of vertical direction, fourth pivot 27 one end with fourth gear 22 coaxial coupling, the 27 other ends of fourth pivot pass in proper order second fixed block 26 the opening 2 is connected with second stirring leaf 28, fourth pivot 27 with second fixed block 26 rotates and connects, barrel 23 one side is provided with feed liquor pipe 29, barrel 23 opposite side is provided with drain pipe 30. The bottom end of the circular ring 19 is provided with a second connecting rod 31 in the vertical direction, the lower end of the second connecting rod 31 penetrates through the opening 2 to be connected with a sweeping brush 32, and bristles of the sweeping brush 32 are in contact with the inner wall of the cylinder 23. The liquid inlet pipe 29 and the liquid outlet pipe 30 are both provided with electromagnetic valves 33. An access hole 34 is formed in the side wall of the transmission case 1, and a sealing plate 35 is arranged at the access hole 34. The sealing plate 35 is connected to the transmission case 1 by screws. And a dust filter 36 is arranged on the outer wall of the transmission case 1. The bottom end of the cylinder 23 is provided with a universal wheel 37.
The working principle and the beneficial technical effects of the technical scheme are as follows: pathogenic escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae are mixed according to a ratio of 4: 4: 2, the solution added with each 1 ten thousand units of streptomycin is fed into the cylinder through the liquid inlet pipe 29, the first motor 3 is started to drive the shift lever 7 to rotate, the shift lever 7 slides in the chute 12 to drive the rack 10 to reciprocate left and right, the second rotating shaft 14 is driven to rotate alternately in forward and reverse rotation through the meshing of the rack 10 and the second gear 13, the two third gears 18 rotate alternately in forward and reverse rotation around the fourth rotating shaft 27, the third gears 18 rotate alternately in forward and reverse rotation due to the meshing of the circular ring 19 and the third gears 18, the fourth gears 22 are driven to rotate alternately in forward and reverse rotation through the meshing of the third gears 18 and the fourth gears 22, the rotation direction of the fourth gears 22 is always opposite to the rotation direction of the third gears 18, correspondingly, the first stirring blade 25 rotates alternately in forward and reverse rotation around the axis of the cylinder 23 and rotates alternately in forward and reverse rotation around the axis of the first stirring blade, and the second stirring blade 28 rotates alternately in forward and reverse rotation around the axis of the second stirring blade, and is always opposite to the rotation direction of the first stirring vane 25, so that a vortex is more easily generated to cause turbulent motion of fluid, and the stirring efficiency is enhanced. In addition, the first gear 6 is meshed with the ring 19 to drive the sweeping brush 32 to rotate continuously, so that the inner wall of the cylinder 23 is swept, and the inner wall of the cylinder is kept clean.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (8)
1. A preparation method of a triple inactivated vaccine for preventing forest musk pneumonia is characterized by comprising the following steps:
step 1, separating pathogenic escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae from pathogenic forest musk deer, and preserving the strains by adopting a freeze drying method;
step 2, respectively placing the three bacteria in a culture medium for culture at 37 ℃, performing amplification culture, and inactivating the bacteria liquid for 24 hours by using formaldehyde with the concentration of 4 per mill;
step 3, diluting each strain to 8.0 × 10 with sterilized normal saline9cfu/mL；
Step 4, mixing pathogenic escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae according to the ratio of 2: 2: mixing the components in a ratio of 1, adding 1 ten thousand units of streptomycin, and uniformly mixing the components by a stirring device to obtain the triple fire-extinguishing vaccine.
2. The method for preparing the triple inactivated vaccine for preventing the woodmusk pneumonia according to claim 1, wherein the stirring device comprises: a transmission case (1), a first motor (3), a first rotating shaft (4), a crank (5), a first gear (6), a deflector rod (7), a guide rail (8), a slide block (9) and a rack (10),
an opening (2) is arranged at the bottom end of the transmission case (1), a first motor (3) is arranged on the bottom wall of the transmission case (1), the output shaft of the first motor (3) is coaxially connected with one end of a first rotating shaft (4), the other end of the first rotating shaft (4) is connected with one end of a crank (5), a first gear (6) is coaxially arranged on the first rotating shaft (4), the other end of the crank (5) is vertically connected with one end of a deflector rod (7), the top of the inner wall of the transmission case (1) is provided with a guide rail (8) in the horizontal direction, the guide rail (8) is connected with a sliding block (9) in a sliding mode, a rack (10) is arranged at the bottom end of the sliding block (9), one end of a first L-shaped connecting rod (11) is connected with one end of the sliding block (9), a sliding groove (12) extending along the length direction is formed in the other end of the first L-shaped connecting rod (11), and the other end of the deflector rod (7) penetrates through the sliding groove (12);
one side of the rack (10) is provided with a second gear (13), the rack (10) is meshed with the second gear (13), a second rotating shaft (14) is coaxially arranged on the second gear (13), one end of the second rotating shaft (14) is connected with the second gear (13), the other end of the second rotating shaft (14) penetrates through a first fixing block (15) and a rotating rod (16) in the horizontal direction to be connected, the second rotating shaft (14) is rotatably connected with the first fixing block (15), the first fixing block (15) is connected with the inner wall of the transmission case (1), two ends of the rotating rod (16) are respectively provided with a first connecting rod (17) in the vertical direction, the lower end of the first connecting rod (17) is coaxially connected with a third gear (18), one side of the first gear (6) is provided with a circular ring (19), the inner ring and the outer ring of the circular ring (19) are both provided with teeth, the outer ring of the circular ring (19) is meshed with the first gear (6), the inner ring of the circular ring (19) is meshed with the two third gears (18) respectively, a sleeve (20) is sleeved on the second rotating shaft (14), the sleeve (20) is rotatably connected with the second rotating shaft (14), second L-shaped rods (21) are arranged on the left side and the right side of the sleeve (20) respectively, one end of each second L-shaped rod (21) is connected with the outer wall of the sleeve (20), the other end of each second L-shaped rod (21) is connected with the upper end of the circular ring (19), a fourth gear (22) is arranged between the two third gears (18), the fourth gear (22) is meshed with the third gears (18), a barrel body (23) is arranged at the bottom end of the transmission case (1), and a third rotating shaft (24) in the vertical direction is arranged at the bottom end of each third gear (18), one end of the third rotating shaft (24) is coaxially connected with the third gear (18), the other end of the third rotating shaft (24) penetrates through the opening (2) to be connected with the first stirring blade (25), a second fixed block (26) is arranged between the fourth gear (22) and the opening (2), the second fixed block (26) is connected with the inner wall of the transmission case (1), a fourth rotating shaft (27) in the vertical direction is arranged at the bottom end of the fourth gear (22), one end of the fourth rotating shaft (27) is coaxially connected with the fourth gear (22), the other end of the fourth rotating shaft (27) sequentially penetrates through the second fixed block (26) and the opening (2) to be connected with a second stirring blade (28), the fourth rotating shaft (27) is rotationally connected with the second fixed block (26), a liquid inlet pipe (29) is arranged on one side of the cylinder body (23), and a liquid outlet pipe (30) is arranged on the other side of the cylinder body (23).
3. The preparation method of the triple inactivated vaccine for preventing the woodmusk pneumonia according to claim 2, wherein a second connecting rod (31) in the vertical direction is arranged at the bottom end of the circular ring (19), the lower end of the second connecting rod (31) penetrates through the opening (2) to be connected with a sweeping brush (32), and bristles of the sweeping brush (32) are in contact with the inner wall of the cylinder body (23).
4. The preparation method of the triple inactivated vaccine for preventing the woodmusk pneumonia according to claim 2, wherein the liquid inlet pipe (29) and the liquid outlet pipe (30) are both provided with electromagnetic valves (33).
5. The preparation method of the triple inactivated vaccine for preventing the woodmusk pneumonia according to the claim 2, characterized in that the transmission case (1) is provided with an access hole (34) on the side wall, and the access hole (34) is provided with a sealing plate (35).
6. The method for preparing the triple inactivated vaccine for preventing the woodmusk pneumonia according to claim 5, wherein the sealing plate (35) is connected with the transmission case (1) through screws.
7. The preparation method of the triple inactivated vaccine for preventing the woodmusk pneumonia according to the claim 5, characterized in that a dust filter screen (36) is arranged on the outer wall of the transmission case (1).
8. The preparation method of the triple inactivated vaccine for preventing the woodmusk pneumonia according to the claim 5, wherein the bottom end of the cylinder body (23) is provided with a universal wheel (37).
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020155128A1 (en) * | 2001-02-04 | 2002-10-24 | Kelly Knape | Inactivated bovine scours vaccines, processes and method of preventing bovine scours |
| CN103948918A (en) * | 2014-03-21 | 2014-07-30 | 母连志 | Method for preparing trivalent inactivated vaccine for preventing mink hemorrhagic pneumonia |
| CN104740622A (en) * | 2015-03-17 | 2015-07-01 | 吉林和元生物工程有限公司 | Pseudomonas aeruginosa, klebsiella and pasteurella triple-inactivated vaccine for mink |
| CN111672383A (en) * | 2020-06-03 | 2020-09-18 | 安徽蜂联生物科技股份有限公司 | A agitating unit for honey processing |
| CN214438411U (en) * | 2021-01-15 | 2021-10-22 | 宜宾市岷源食品有限公司 | A agitating unit for sausage processing |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020155128A1 (en) * | 2001-02-04 | 2002-10-24 | Kelly Knape | Inactivated bovine scours vaccines, processes and method of preventing bovine scours |
| CN103948918A (en) * | 2014-03-21 | 2014-07-30 | 母连志 | Method for preparing trivalent inactivated vaccine for preventing mink hemorrhagic pneumonia |
| CN104740622A (en) * | 2015-03-17 | 2015-07-01 | 吉林和元生物工程有限公司 | Pseudomonas aeruginosa, klebsiella and pasteurella triple-inactivated vaccine for mink |
| CN111672383A (en) * | 2020-06-03 | 2020-09-18 | 安徽蜂联生物科技股份有限公司 | A agitating unit for honey processing |
| CN214438411U (en) * | 2021-01-15 | 2021-10-22 | 宜宾市岷源食品有限公司 | A agitating unit for sausage processing |
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