Haemophilus parasuis disease and swine pasteurella multocida disease combined vaccine and preparation method thereof
A technology for Haemophilus suis disease and Haemophilus suis, applied in antibacterial drugs, pharmaceutical formulations, bacterial antigen components, etc., can solve problems such as different immune procedures, and achieve the effect of long immune protection period and good immune effect.
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Embodiment 1
[0029] Embodiment 1: the preparation of double vaccine
[0030] 1. Preparation of seeds for production
[0031] Propagation of primary seeds: Haemophilus parasuis serotype 4 JS strain and type 5 ZJ strain freeze-dried strains were streaked and inoculated on TSA / NAD (TSA was produced by BD Company, NAD was produced by Roche Company) plates, Cultivate at 37°C for 18-24 hours, select colonies that meet the requirements, inoculate them in TSB / NAD (TSB is produced by BD Company, NAD is produced by Roche Company) liquid medium, and cultivate at 37°C for 12-16 hours, as the primary seed ; Pasteurella multocida type A HN5 strain, Pasteurella multocida type B type C44-1 strain and Pasteurella multocida type D type HB4 strain freeze-dried strains were streaked and inoculated in TSA (10% chicken serum ) (chicken serum was produced by Zhengzhou Bai'an Bioengineering Co., Ltd.) plates, and cultured at 37°C for 18-24 hours as first-class seeds.
[0032] Propagation of secondary seeds: Tak...
Embodiment 2
[0044] Embodiment 2: Sterility test and safety test of dual vaccine
[0045] 1. Sterility test
[0046] According to the method on page 50 of the appendix of "Regulations of Veterinary Biological Products of the People's Republic of China" 2000 edition, the vaccine prepared by the present invention was sterility tested; the result was that no colony growth was seen in the vaccine obtained.
[0047] 2. Safety test
[0048] Safety test on mice: Select 10 3-week-old BALB / c mice and divide them into two groups, 5 mice in each group. One group is injected with 0.5ml of this inactivated vaccine subcutaneously, and the other group is injected with 0.1ml in the thigh muscle. Observation after 10 days. As a result, the mice were in a good state of mind, with no adverse reactions and no deaths.
[0049] Piglet safety test: select 10 healthy susceptible piglets within 30 days of age, randomly select 5 piglets for neck intramuscular injection with the double dose according to usage (th...
Embodiment 3
[0051] Embodiment 3: Efficacy test of dual vaccine
[0052] 50 healthy and susceptible 3-week-old weaned piglets were selected and divided into two groups, 25 in the immunization group and 25 in the control group. 3-week-old piglets were immunized for the first time, with an interval of 2 weeks, and booster immunization was carried out at the age of 5 weeks, with an interval of 2 weeks, and each virulent strain was used to challenge the virus at the age of 7 weeks; the vaccine dosage for each immunization was 2ml. The finished product of the combined vaccine; after the challenge, observe the statistical results for 14 days.
[0053] Haemophilus parasuis JS strain: Select 5 immunized pigs, together with 5 unimmunized pigs with the same conditions, intraperitoneally inject Haemophilus parasuis JS strain, the challenge dose is 5.0×10 9 CFU / head, observed for 14 days after challenge, more than 4 / 5 of the immune piglets were protected, and more than 4 / 5 of the control piglets were...
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