猪链球菌3型疫苗候选菌株筛选及3＋9型二价灭活疫苗对小鼠免疫效果评估

doi:10.16431/j.cnki.1671-7236.2022.06.031

Abstract

Abstract: 【Objective】 The study was aimed to screen vaccine candidate strains of Streptococcus suis serotype 3, prepare a bivalent inactivated vaccine of Streptococcus suis serotype 3+9 and evaluate its immunoprotective effect on BALB/c mice.【Method】 In this study, serotype 3 strain of Streptococcus suis isolated from diseased pigs were screened by the larvae of Galleria mellonella and BALB/c mice model and was studied as vaccine candidate strains.Then the biological characteristics of the candidate strains were studied.The serotype 3 vaccine candidate strains and the previously screened serotype 9 candidate vaccine strains were inactivated and concentrated, and the antigen concentration was 2×10¹⁰ CFU/mL.After the sterility test, the concentrated antigen liquid was mixed in a ratio of 1:1, and then mixed with the Summit Poly Solution adjuvant in a ratio of 4:1 to prepare bivalent inactivated vaccine of Streptococcus suis serotype 3+9, the bacterial content of Streptococcus suis serotype 3 and 9 were both 2×10⁹ CFU/mL.The prepared vaccine was inoculated into 6-week-old BABL/c mice.The second immunization was carried out on the 14th day after the first immunization, and the challenge was carried out on the 14th day after the second immunization.At the same time, a commercial vaccine immunization group and a negative control group were established to evaluate the safety and immune protection effect of the vaccine.【Result】 PCR identification result showed that all 23 clinical isolates were Streptococcus suis serotype 3, named KQ3-1 to KQ3-23 in succession.The virulent strain KQ3-1 was screened by primary screening and rescreening of Galleria mellonella and BALB/c mice.Virulence gene testing showed that the genotype of KQ3-1 was gapdh⁺/sly⁺/fbps^-/orf2^-/mrp^-/89K^-/gdh⁺/epf^-.The growth curve showed that the strain reached its peak when cultured at 37 ℃ for 8 to 10 h.The pathogenicity of BALB/c mice showed that intraperitoneal inoculation could cause clinical symptoms such as depression, pile up, hair erection, motor retardation and death within 12 h.The LD₅₀ of the strain was 5.2×10⁷ CFU/piece.After immunizing the mice with the prepared vaccine, the mental status and food intake of the mice were normal, there were no adverse reactions such as swelling and lumps at the injection site of the vaccine, and there was no death, indicating that the vaccine had good safety.After immunization, Streptococcus suis serotype 2 strain ZYS, Streptococcus suis serotype 3 strain KQ3-1 and Streptococcus suis serotype 9 strain YT were challenged.The mortality rate of control group was 90%, 100% and 100%, respectively.The protection rates of Streptococcus suis serotype 3+9 bivalent inactivated vaccine immunization group were 30%, 80% and 70%, respectively.The protection effects of commercial vaccine group were 70%, 0 and 10%, respectively.【Conclusion】 The vaccine developed in this study could provide good immune protection against Streptococcus suis serotype 3 and 9 strains and had potential value of vaccine market development.

Key words: Streptococcus suis; serotype 3 and 9; bivalent inactivated vaccine

CLC Number:

S852.61⁺1

LI Qianqian, LONG Yunzhi, LIANG Gong, JIN Qing, LIU Jinjin, YANG Liu, HUANG Ying, YU Daobing, SONG Wenbo, HUANG Chao, TANG Xibiao. Screening of Vaccine Candidate Strains of Streptococcus suis Serotype 3 and Evaluation of the Immune Effect of the Bivalent Inactivated Vaccine Streptococcus suis Serotype 3+9 on Mice[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2307-2317.

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Screening of Vaccine Candidate Strains of Streptococcus suis Serotype 3 and Evaluation of the Immune Effect of the Bivalent Inactivated Vaccine Streptococcus suis Serotype 3+9 on Mice

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