Acta Veterinaria et Zootechnica Sinica ›› 2024, Vol. 55 ›› Issue (9): 4077-4090.doi: 10.11843/j.issn.0366-6964.2024.09.032
• Preventive Veterinary Medicine • Previous Articles Next Articles
Lu PENG(), Heng ZHANG(), Siqi PANG, Zhulin QIAO, Xiaofen ZHANG, Chen TAN, Yunfeng SONG, Rui ZHOU, Lu LI*()
Received:
2023-11-20
Online:
2024-09-23
Published:
2024-09-27
Contact:
Lu LI
E-mail:penglu1998@webmail.hzau.edu.cn;zhangheng666@webmail.hzau.edu.cn;lilu@mail.hzau.edu.cn
CLC Number:
Lu PENG, Heng ZHANG, Siqi PANG, Zhulin QIAO, Xiaofen ZHANG, Chen TAN, Yunfeng SONG, Rui ZHOU, Lu LI. Screening of Trivalent Inactivated Vaccine Candidate Strains of Streptococcus suis Serotype 2, 3 and 9 Using Galleria mellonella and Mice Infection Models[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4077-4090.
Table 1
PCR primers"
引物Primers | 引物序列(5′→3′) Sequence | 产物大小/bp Product |
cps2I-F | TTCGTATTAACTTACTTGGCGT | 363 |
cps2I-R | TAAATCCCCATATGCCAAATCC | |
cps3L-F | ACATCCATTGCAGGAGTAGT | 210 |
cps3L-R | TGCAGTTCCAAAATTCTTCGT | |
cps9J-F | TCCCTTTAATTTCACAGGCAAT | 409 |
cps9J-R | CTGTCAAAGAATTGAATCCCAC | |
gapdh-F | CACACTTGTTGAAATATGACAC | 571 |
gapdh-R | AGCACCGTCAAGTTTACC | |
sly-F | GCAGAAGGGACAACTGTAGAAG | 443 |
sly-R | GCACTGCTATTATGGACTGTTGA | |
fbps-F | CAAGGTTTGGGTCGGGATA | 720 |
fbps-R | CCCGTCTGTTGCCAAGTAA | |
orf2-F | CAAGTGTATGTGGATGGG | 858 |
orf2-R | ATCCAGTTGACACGTGCA | |
mrp-F | GGTATACCTTGCTGGTACGGTTC | 532 |
mrp-R | AGTCTCTACAGCTGTAGCTGG | |
89K-F | GATATCGCCACTATGGTATC | 720 |
89K-R | GATTGTGGACCATGCTCTTTAG | |
gdh-F | GCAGCGYAYYCYGYCAAACG | 688 |
gdh-R | CCATGGACAGATAAAGATGG | |
epf-F | ACAAAGGCGTAGGTTCAATC | 269 |
epf-R | CGGCATCAAGAATGTCTTTG |
Table 2
Inactivated vaccine immunization program and immune dose"
组别Group | 免疫剂量Immune dose | 数量Quantity | 免疫方式Vaccination method | 攻毒菌株/剂量Challenge strain/dose | 攻毒方式Challenge method |
SS1803免疫组 SS1803 | 4×107 CFU | 6 | 肌肉注射Intramuscular injection | SS1803: 5×107 CFU | 腹腔注射Intraperitoneal injection |
SS1803免疫组 SS1803 | 8×107 CFU | 6 | 肌肉注射Intramuscular injection | SS1803: 5×107 CFU | 腹腔注射Intraperitoneal injection |
SS1803024免疫组 SS1803024 | 2×108 CFU | 6 | 肌肉注射Intramuscular injection | SS1803024: 1.7×108 CFU | 腹腔注射Intraperitoneal injection |
SS1803024免疫组 SS1803024 | 5×108 CFU | 6 | 肌肉注射Intramuscular injection | SS1803024: 1.7×108 CFU | 腹腔注射Intraperitoneal injection |
SS1696免疫组 SS1696 | 1×108 CFU | 6 | 肌肉注射Intramuscular injection | SS1696: 1.5×107 CFU | 腹腔注射Intraperitoneal injection |
SS1696免疫组 SS1696 | 5×108 CFU | 6 | 肌肉注射Intramuscular injection | SS1696: 1.5×107 CFU | 腹腔注射Intraperitoneal injection |
三价苗免疫组 TIV | 2×107 CFU SS1803+1.5×108 CFU SS1803024+3×107 CFU SS1696 | 18 | 肌肉注射Intramuscular injection | SS1803: 5×107 CFU SS1803024: 1.7×108 CFU SS1696: 1.5×107 CFU | 腹腔注射Intraperitoneal injection |
阴性对照组 Negative | PBS | 18 | 肌肉注射Intramuscular injection | SS1803: 5×107 CFU SS1803024: 1.7×108 CFU SS1696: 1.5×107 CFU | 腹腔注射Intraperitoneal injection |
Table 3
Screening results of S.suis serotype 3, 9 strains on Galleria mellonella %"
血清3型菌株Serotype 3 strains | 第一轮死亡率(n=10) Mortality rate of first round experiment | 第二轮死亡率(n=10) Mortality rate of second round experiment | 血清9型菌株Serotype 9 strains | 第一轮死亡率(n=10) Mortality rate of first round experiment | 第二轮死亡率(n=10) Mortality rate of second round experiment | |
SS1761 | 60 | 80 | SS1802036 | 60 | 50 | |
SS1804125 | 100 | 60 | SS1807065 | 80 | 80 | |
SS1730 | 10 | 10 | SS1823 | 30 | 40 | |
SS1767 | 20 | 20 | SS1805031 | 90 | 70 | |
SS1766 | 30 | 40 | SS1805032 | 60 | 40 | |
SS1968 | 10 | 10 | SS1804104 | 90 | 90 | |
SS1803024 | 100 | 60 | SS1804097 | 30 | 20 | |
SS1803025 | 70 | 60 | SS1407 | 0 | 10 | |
SS2214 | 30 | 10 | SS1904022 | 10 | 10 | |
SS1703 | 40 | 30 | SS1681 | 40 | 20 | |
SS1804072 | 40 | 10 | SS1696 | 80 | 80 | |
SS1808002 | 30 | 20 | SS1697 | 70 | 60 | |
生理盐水组Normal saline group | 0 | 0 | SS1807038 | 40 | 20 | |
SS1807035 | 80 | 80 | ||||
空白对照组Blank control group | 0 | 0 | SS1807091 | 30 | 30 | |
SS1807090 | 30 | 0 | ||||
生理盐水组Normal saline group | 0 | 0 | ||||
空白对照组Blank control group | 0 | 0 |
Fig. 2
Growth characteristics and identification of virulence factors of SS serotype 2, 3 and 9 strains A, D, G. OD600 nm value of SS1803, SS1803024, SS1696; B, E, H. Bacterial count of SS1803, SS1803024, SS1696; C, F, I. Virulence factor identification of SS1803, SS1803024, SS1696: M. 5 000 DNA marker; 1. gapdh gene; 2. sly gene; 3. fbps gene; 4. orf2 gene; 5. mrp gene; 6. 89K gene; 7. gdh gene; 8. epf gene"
Fig. 7
Determination of inflammatory cytokines in serum of mice at 12 and 24 h after challenge(n=6) A, B, C. IL-6 levels in serum of mice after SS1803、SS1803024、SS1696 challenge; D, E, F. IL-12 levels in serum of mice after SS1803, SS1803024, SS1696 challenge. *. P < 0.05; **. P < 0.01; ***. P < 0.001; ****. P < 0.000 1;ns. P > 0.05"
Fig. 8
Pathological histological sections of brain, spleen, lung and kidneys of mice after challenge(200× and 100×) A. Control group of SS1803 challenge; B. TIV group of SS1803 challenge; C. Control group of SS1803024 challenge; D. TIV group of SS1803024 challenge; E. Control group of SS1696 challenge; F. TIV group of SS1696 challenge; G. Blank group(bar: 200 μm, 100 μm). Brain tissue sections: red arrows indicate neuronal cell degeneration with pyknotic hyperchromatic nuclei; green arrows indicate mild hemorrhage in brain parenchyma. Spleen tissue sections: red arrows indicate increased multinucleated macrophages; green arrows indicate a small amount of hemosiderin deposition. Lung tissue sections: red arrows indicate alveolar epithelial cell hyperplasia; green arrows indicate a small amount of protein mucus in the bronchial lumen; yellow arrows indicate inflammatory cell infiltration. Kidney tissue sections: red arrows indicate glomerular lobulation; black arrows indicate atrophy and loss of some glomeruli in the cortical area, and the number of mesangial cells in the glomerulus is reduced"
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