CN-121987770-A - Five-valent inactivated vaccine for infectious serositis of duck and preparation method and application thereof

Abstract

The invention relates to a pentavalent inactivated vaccine for infectious serositis of ducks, and a preparation method and application thereof. Is prepared from infectious serositis 1 antigen, infectious serositis 2 antigen, infectious serositis 6 antigen, infectious serositis 7 antigen and infectious serositis 10 antigen. The preparation method comprises the steps of S1, bacterial strain screening, S2, bacterial strain culturing, S3, inactivation treatment, S4, bacterial liquid mixing, S5 and adjuvant emulsification. The five-valent inactivated vaccine for infectious serositis of ducks has the advantages of prominent broad spectrum, excellent immune effect and high industrialized feasibility, can cover more than 90% of epidemic strains, solves the problem of incomplete prevention of the existing vaccine, has protection rate of greater than or equal to 90% on virulent attack of five serotypes, adopts a fermentation tank for enlarged culture in a culture process, has stable bacterial liquid proliferation efficiency and definite emulsification process parameters, and is suitable for mass production.

Inventors

• LI E
• LIU XINGGANG
• QI DAN
• CUI SONGQI
• HAN ZHIQIANG
• CHAI PEIYAO
• Wen Xingying
• WANG FUCHAO
• ZHANG LIANG
• ZHANG NA
• JIANG DONG

Assignees

• 豪威生物科技有限公司

Dates

Publication Date

20260508

Application Date

20251216

Claims (4)

1. 1. The five-price inactivated vaccine for the duck infectious serositis is characterized by being prepared from duck infectious serositis 1 antigen, duck infectious serositis 2 antigen, duck infectious serositis 6 antigen, duck infectious serositis 7 antigen and duck infectious serositis 10 antigen, wherein the duck infectious serositis 1 antigen is HVJZ-08001 antigen, the viable count of the antigen before concentration is 200-300 CFU/mL, the duck infectious serositis 2 antigen is HVJZ-08002 antigen, the viable count of the antigen before concentration is 300-400 CFU/mL, the duck infectious serositis 6 antigen is JN21061 three antigen strains, the viable count of the antigen before concentration is 300-400 CFU/mL, the duck infectious serositis 7 antigen is GX21070 antigen, the viable count of the antigen before concentration is 450-550 CFU/mL, the duck infectious serositis 10 antigen is SD211001 antigen, and the viable count of the antigen before concentration is 400-500 CFU/mL.
2. 2. The pentavalent inactivated vaccine for duck infectious serositis of claim 1, wherein the viable count of unconcentrated pre-antigen for duck infectious serositis type 1 antigen is 250CFU/mL, the viable count of unconcentrated pre-antigen for duck infectious serositis type 2 antigen is 350CFU/mL, the viable count of unconcentrated pre-antigen for duck infectious serositis type 6 antigen is 350CFU/mL, the viable count of unconcentrated pre-antigen for duck infectious serositis type 7 antigen is 500CFU/mL, and the viable count of unconcentrated pre-antigen for duck infectious serositis type 10 antigen is 450CFU/mL.
3. 3. The method for preparing the pentavalent inactivated vaccine for infectious serositis of duck of claim 2, comprising the following steps, Step S1, screening strains; Selecting duck infectious serositis 1 antigen HVJZ and 08001, duck infectious serositis 2 antigen HVJZ and 08002, duck infectious serositis 6 antigen JN21061, duck infectious serositis 7 antigen GX21070 and duck infectious serositis 10 antigen SD211001, and carrying out microorganism identification and confirmation on each strain and freezing each strain; step S2, strain culture; Preparing a special culture medium, adjusting the pH value to 6.8-7.0, and adding 5% serum for later use; Taking frozen and preserved strains, respectively inoculating each strain to a special culture medium, culturing at a constant temperature of 37 ℃ for 6-16 hours, then inoculating the first-generation bacterial liquid of each strain to the special culture medium according to 5%, culturing at a constant temperature of 37 ℃ and detecting an OD value, inoculating the second-stage bacterial liquid to the special liquid culture medium according to 5%, respectively inoculating the second-stage bacterial liquid to a fermentation tank, culturing at a constant temperature of 37 ℃ with DO value of 30-50%, rotating speed of 200rpm and ventilation rate of 2Lpm, stopping culturing until the OD 600 value of the bacterial liquid reaches more than 1.5, and detecting the OD value and viable bacteria count; centrifuging the culture solution to collect the antigen; s3, inactivating treatment; Respectively concentrating the obtained antigen 10 times, inactivating with formaldehyde with final concentration of 2%o at 37deg.C for 24 hr, collecting inactivated bacteria liquid 0.5mL, inoculating the inactivated bacteria liquid of five strains with TSA slant 2 branch containing 2% new born calf serum, culturing at 37 deg.C for 7d, and observing whether bacteria grow; s4, mixing bacterial liquid; Mixing five inactivated bacterial solutions of duck infectious serositis 1-type antigen HVJZ08001 strain, duck infectious serositis 2-type antigen HVJZ08002 strain, duck infectious serositis 6-type antigen JN21061 strain, duck infectious serositis 7-type antigen GX21070 strain and duck infectious serositis 10-type antigen SD211001 strain according to the volume ratio of 1-3:1-3, and stirring for 30min to prepare a mixed bacterial solution; step S5, emulsifying an adjuvant; preparing a water phase and an oil phase respectively, taking 2 parts of the oil phase, placing the oil phase in an emulsifying tank, slowly adding 1 part of the water phase, emulsifying for 10min at 8000rpm, taking 10mL after emulsification, centrifuging for 15min at 3000rpm, quantitatively packaging and sealing with a cover, and preserving at 2-8 ℃.
4. 4. Use of the pentavalent inactivated vaccine for duck infectious serositis according to claim 2 for immunization and treatment of duck infectious serositis.

Description

Five-valent inactivated vaccine for infectious serositis of duck and preparation method and application thereof Technical Field The invention belongs to the technical field of poultry vaccines, in particular to a pentavalent inactivated vaccine for infectious serositis of ducks, and also designs a preparation method and application of the vaccine, which are suitable for immune protection from duckling to adult ducks and can prevent and control epidemic of the infectious serositis of multi-serotype ducks. Background The infectious serositis of the ducks is an acute and contact infectious disease caused by the riemerella anatipestifer, and mainly damages duckling of 1-8 weeks old, the death rate reaches 30-70%, and the economic benefit of the duck industry is seriously influenced. The current core means for controlling the epidemic disease in the industry is vaccination, but the prior art has the following key defects: 1. the serotype coverage is incomplete, the current commercial vaccine only aims at 2 serotypes, and the Riemerella anatipestifer has confirmed that the serotypes are more than 20, and recent epidemiological investigation in China shows that the epidemic rate of the serotypes of 6 types, 7 types and 10 types in main culture areas such as Jiangsu, shandong and Sichuan is 45-60%, and the existing bivalent vaccine cannot cover the emerging epidemic serotypes. 2. The cross protection capability is lacking, the capsular polysaccharide antigens of different serotypes of riemerella anatipestifer are obviously different, and the existing bivalent vaccine does not have cross protection on type 6, 7 and 10 strains, so that the vaccinated duck group is still susceptible to disease. 3. The immunization program is complex and has high cost, part of farms are covered with multiple serotypes, and monovalent vaccines of type 1, type 2, type 6 and the like are respectively inoculated, so that the immunization times are increased, the labor cost is increased, the weight gain rate of meat ducks is reduced due to stress of duck groups caused by frequent duck grabbing, and the laying rate of the laying ducks is reduced. In summary, the prior art cannot meet the industry requirement of "one-seedling prevention multiple types, low cost and high efficiency prevention and control", and development of broad-spectrum vaccines covering the dominant epidemic serotypes is urgently needed. Disclosure of Invention The invention aims to provide a pentavalent inactivated vaccine for infectious serositis of ducks, which realizes synchronous coverage of 1-type, 2-type, 6-type, 7-type and 10-type serovars of the infectious serositis of ducks and solves the problems of insufficient coverage of the serovars, lack of cross protection capability, complex immunization program and high cost of the existing vaccine. The invention adopts the technical scheme that the five-valent inactivated vaccine for duck infectious serositis is prepared from duck infectious serositis 1 antigen, duck infectious serositis 2 antigen, duck infectious serositis 6 antigen, duck infectious serositis 7 antigen and duck infectious serositis 10 antigen, wherein the duck infectious serositis 1 antigen is HVJZ08001 strain antigen, the viable count of the unconcentrated pre-antigen is 200-300 CFU/mL, the duck infectious serositis 2 antigen is HVJZ08002 strain antigen, the viable count of the unconcentrated pre-antigen is 300-400 CFU/mL, the duck infectious serositis 6 antigen is JN21061 three strain, the viable count of the unconcentrated pre-antigen is 300-400 CFU/mL, the duck infectious serositis 7 antigen is GX21070 three strain antigen, the viable count of the unconcentrated pre-antigen is 450-550 CFU/mL, the infectious serositis 10 antigen is 211001 strain antigen, and the viable count of the unconcentrated pre-antigen is 400-500 CFU/mL. Preferably, the pre-concentration live pre-antigen count of the duck infectious serositis type 1 antigen is 250CFU/mL, the pre-concentration live pre-antigen count of the duck infectious serositis type 2 antigen is 350CFU/mL, the pre-concentration live pre-antigen count of the duck infectious serositis type 6 antigen is 350CFU/mL, the pre-concentration live pre-antigen count of the duck infectious serositis type 7 antigen is 500CFU/mL, and the pre-concentration live pre-antigen count of the duck infectious serositis type 10 antigen is 450CFU/mL. The invention also aims to provide a preparation method of the pentavalent inactivated vaccine for infectious serositis of ducks, which comprises the following steps: Step S1, screening strains; Selecting duck infectious serositis 1 antigen HVJZ and 08001, duck infectious serositis 2 antigen HVJZ and 08002, duck infectious serositis 6 antigen JN21061, duck infectious serositis 7 antigen GX21070 and duck infectious serositis 10 antigen SD211001, and carrying out microorganism identification and confirmation on each strain and freezing each strain; step S2, strain culture; Preparing a sp