斯氏艾美耳球虫重组表面抗原SAG13和SAG14对兔的免疫保护效果初步观察

doi:10.11843/j.issn.0366-6964.2022.03.021

摘要/Abstract

摘要： 本研究旨在观察斯氏艾美耳球虫裂殖生殖阶段特异性表面抗原Es-SAG13和Es-SAG14的原核表达产物对兔的免疫保护效果。利用相对荧光定量PCR分析Es-SAGs在未孢子化卵囊时期、孢子化卵囊时期、裂殖生殖时期和配子生殖时期的转录水平,并对裂殖生殖阶段特异性表达的Es-SAG进行原核表达;然后将40只45日龄无球虫幼兔随机分为4组,分别是空白对照组、攻虫对照组、rEs-SAG13免疫组和rEs-SAG14免疫组,分别经颈部皮下注射1 mL PBS、1 mL PBS、1 mL (100 μg·mL^-1) rEs-SAG13、1 mL (100 μg·mL^-1) rEs-SAG14,首免14 d后,同等剂量加强免疫,二免14 d后,除空白对照外经口感染1×10⁴个斯氏艾美耳球虫孢子化卵囊·只^-1,观察各组临床症状并于感染21 d后剖检,并统计各组的卵囊排出量、相对增重、料肉比、肝指数和肝病变记分。结果显示:Es-SAGs在各阶段的转录水平存在差异,其中,SAG13、SAG14属于裂殖生殖阶段特异表达的成员,原核表达的rEs-SAG13和rEs-SAG14均具有良好的反应原性。免疫保护试验表明:感染后攻虫对照组出现兔肝球虫病典型症状,而免疫组症状不明显;rEs-SAG13免疫组和rEs-SAG14免疫组的卵囊减少率分别达82.8%和51.9%;免疫组平均相对增重均显著大于攻虫对照组(P<0.05);rEs-SAG13免疫组与攻虫对照组的肝指数、肝病变记分差异均显著(P<0.05),而rEs-SAG14免疫组与攻虫对照组的肝指数、肝病变记分差异均不明显(P>0.05)。rEs-SAG13和rEs-SAG14均能不同程度地减少增重损失和卵囊排出,减轻肝病变,其中rEs-SAG13免疫保护效果更佳。

关键词: 斯氏艾美耳球虫, 表面抗原, SAG13, SAG14, 原核表达, 免疫保护

Abstract: This experiment was conducted to observe the immunoprotective effects of the prokaryotic expression products of surface antigens SAG13 and SAG14 of Eimeria stiedae in rabbits. We analyzed the transcription level of Es-SAGs in unsporulated oocyst stage, sporulated oocyst stage, schizogony stage and gametogony stage by relative fluorescence quantitative PCR, and expressed Es-SAG specifically expressed in the schizogony stage. Then forty 45-day-old coccidia-free rabbits were randomly divided into four groups:blank control group (B), parasite challenge control group (P), rEs-SAG13 immunization group (SAG13) and rEs-SAG14 immunization group (SAG14). Rabbits of group B, group P, group SAG13 and group SAG14 were subcutaneously injected with 1 mL PBS, 1 mL PBS, 1 mL 100 μg·mL^-1 rEs-SAG13 and 1 mL 100 μg·mL^-1 rEs-SAG14, respectively, then the same amount of booster immunity was given after 14 days. Two weeks after the second immunization, 1×10⁴ sporulated oocysts were infected orally in all groups except group B. Three weeks after infection, the differences of clinical symptoms, oocyst excretion, relative weight gain, feed conversion ratio, liver index and liver lesions were observed in each group. Results revealed that the transcription level of Es-SAGs at each stage is significantly different, and SAG13 and SAG14 are members that are specifically expressed in schizogony stage. The rEs-SAG13 and rEs-SAG14 have good reactogenicity. The typical symptoms of rabbit hepatic coccidiosis appeared in the control group, while the symptoms of the other groups were not obvious. The oocyst reduction rates of rEs-SAG13 immunization group and rEs-SAG14 immunization group were 82.8% and 51.9%, respectively. The average relative weight gain of immune group was significantly higher than that of parasite challenge control group. There were significant differences in liver index and liver lesion score between rEs-SAG13 immunization group and parasite challenge control group. However, there was no significant difference in liver index and liver lesion score between rEs-SAG14 immunization group and parasite challenge control group (P<0.05). These results indicated that rEs-SAG13 and rEs-SAG14 can reduce weight gain loss and oocyst output in varying degrees, alleviate liver lesions, but rEs-SAG13 is more effective.

Key words: Eimeria stiedae, surface antigens, surface antigen 13, surface antigen 14, prokaryotic expression, immunoprotective

中图分类号:

S852.723

罗跃军, 任永军, 白鑫, 陈浩, 蒲家艳, 郑若愚, 肖洁, 杨光友. 斯氏艾美耳球虫重组表面抗原SAG13和SAG14对兔的免疫保护效果初步观察[J]. 畜牧兽医学报, 2022, 53(3): 883-893.

LUO Yuejun, REN Yongjun, BAI Xin, CHEN Hao, PU Jiayan, ZHENG Ruoyu, XIAO Jie, YANG Guangyou. Preliminary Observation on the Immunoprotective Effects of Recombinant Surface Antigens SAG13 and SAG14 of Eimeria stiedae in Rabbits[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 883-893.

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