巨型艾美耳球虫与产气荚膜梭菌共感染致鸡坏死性肠炎模型的评价

doi:10.11843/j.issn.0366-6964.2025.08.049

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (8): 4120-4128.doi: 10.11843/j.issn.0366-6964.2025.08.049

• 研究简报 • 上一篇

巨型艾美耳球虫与产气荚膜梭菌共感染致鸡坏死性肠炎模型的评价

袁橙¹(), 袁月²(), 张清正², 宋小凯², 徐立新², 严若峰², 李祥瑞², 陆明敏²^,*()

1. 江苏农牧科技职业学院, 泰州 225300
2. 南京农业大学动物医学院, 南京 210095

收稿日期:2025-02-26 出版日期:2025-08-23 发布日期:2025-08-28
通讯作者: 陆明敏 E-mail:840289909@qq.com;1290721798@qq.com;mingmin.lu@njau.edu.cn
作者简介:袁橙(1986-)，女，江苏盐城人，博士，副教授，主要从事预防兽医学、职业教育研究，E-mail: 840289909@qq.com
袁月(2000-)，女，辽宁抚顺人，硕士，主要从事预防兽医学、兽医寄生虫学研究，E-mail: 1290721798@qq.com
第一联系人：
袁橙和袁月为同等贡献作者
YUAN Cheng and YUAN Yue Contributed equally to this work.
基金资助:
国家自然科学基金青年基金(32202837)；江苏农牧科技职业学院校级科研项目(NSF2023ZR02)；江苏高校“青蓝工程”(苏教师函〔2024〕14号)

Evaluation of a Necrotic Enteritis Model Induced by Co-infection with Eimeria maxima and Clostridium perfringens

YUAN Cheng¹(), YUAN Yue²(), ZHANG Qingzheng², SONG Xiaokai², XU Lixin², YAN Ruofeng², LI Xiangrui², LU Mingmin²^,*()

1. Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China
2. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China

Received:2025-02-26 Online:2025-08-23 Published:2025-08-28
Contact: LU Mingmin E-mail:840289909@qq.com;1290721798@qq.com;mingmin.lu@njau.edu.cn

摘要/Abstract

摘要：

鸡球虫感染破坏宿主肠道免疫稳态，是引起产气荚膜梭菌(Clostridium perfringens, CP)介导的鸡坏死性肠炎(necrotic enteritis，NE)的主要诱因。在7种鸡球虫中，巨型艾美耳球虫(Eimeria maxima，EM)最适于诱导NE。因此，本研究旨在通过对不同EM攻虫剂量的探究，建立稳定、可靠的NE实验室模型。将200只14日龄的肉鸡分为10组，分别为空白对照组、CP攻毒组、EM攻虫组(不同攻虫剂量)、EM/CP共感染组(不同攻虫剂量)。在14日龄时，使用不同剂量E. maxima孢子化卵囊攻虫。在感染后4 d使用C. perfringens攻毒，在20日龄时终止试验，剖杀鸡只，记录攻虫前、20日龄体重及存活率并进行肠道病变计分。结果显示，随着E. maxima感染剂量的增加，单一E. maxima感染组的增重逐渐减缓，但在EM/CP共感染组未见该趋势，EM/CP1组中相对增长率显著高于其余EM/CP共感染组，EM/CP2组、EM/CP3组、EM/CP4组间未见显著差异。肠道病变记分呈现与体重增长率较为相似的趋势，EM/CP1组肠道病变程度较其他EM/CP共感染组较轻，且EM/CP2组、EM/CP3组、EM/CP4组间未见显著区别。在EM/CP共感染组中，EM/CP2组的存活率为85%，而EM/CP3组和EM/CP4组存活率偏低，分别为80%和70%。综合增重情况、肠道病变、致死率三个指标进行判断，EM/CP2组的攻虫攻毒剂量(1×10⁴卵囊·羽^-1+1×10⁹ CFUs·羽^-1)最适用于E. maxima诱发NE的实验室模型的构建，该模型可为NE的致病机制研究、疫苗及药物研发等提供较好的研究工具。

关键词: 产气荚膜梭菌, 巨型艾美耳球虫, 鸡坏死性肠炎, 双重感染模型

Abstract:

Coccidiosis disrupts the intestinal immune homeostasis of the host, serving as a major predisposing factor for Clostridium perfringens (CP)-mediated necrotic enteritis (NE). Among seven chicken Eimeria spp., Eimeria maxima (EM) is the most effective at inducing NE. Accordingly, the objective of this study is to establish a stable and reliable experimental model of NE by evaluating the impact of varying EM challenge doses. A total of 200 14-day-old broilers were divided into 10 groups: the control group, CP challenge group, EM infection groups (with varying infection doses), and EM/CP co-infection groups (with varying infection doses). At 14 days of age, different doses of EM sporulated oocysts were inoculated. 4 days post EM infection, CP isolates were inoculated for the challenge infection. The trial was terminated on Day 20, and all the chickens were sacrificed. The body weights (before the challenge and on Day 20) and the survival rate were recorded, and the lesions of the small intestines were scored. The results showed that with the increase of EM infection dose, the weight gain was gradually reduced in the single EM infection groups, whereas this trend was not observed in the EM/CP co-infection groups. The relative weight gain of the EM/CP1 group was significantly higher than those of the other EM/CP co-infection groups. However, no significant changes were observed across the EM/CP2, EM/CP3, and EM/CP4 groups. The lesion scores in the challenged groups showed a trend similar to the relative weight gains. The intestinal lesions in the EM/CP1 group were less severe than those in the other EM/CP co-infection groups, and there was no significant difference in gut lesions among the EM/CP2, EM/CP3, and EM/CP4 groups. In the EM/CP co-infection groups, the survival rate of the EM/CP2 group was 85%, while the survival rates of the EM/CP3 and EM/CP4 groups were lower, at 80% and 70%, respectively. Based on the comprehensive evaluation of the relative weight gains, intestinal lesions, and mortality, the EM/CP2 group dosage (1×10⁴ oocysts/bird + 1×10⁹ CFUs/bird) was determined to be the optimal infection dose for inducing an experimental model of NE. Consequently, this model provides a valuable research tool for the investigation of NE pathogenesis and the development of vaccines/drugs against NE.

Key words: C. perfringens, Emaxima, necrotic enteritis, dual infection model

中图分类号:

袁橙, 袁月, 张清正, 宋小凯, 徐立新, 严若峰, 李祥瑞, 陆明敏. 巨型艾美耳球虫与产气荚膜梭菌共感染致鸡坏死性肠炎模型的评价[J]. 畜牧兽医学报, 2025, 56(8): 4120-4128.

YUAN Cheng, YUAN Yue, ZHANG Qingzheng, SONG Xiaokai, XU Lixin, YAN Ruofeng, LI Xiangrui, LU Mingmin. Evaluation of a Necrotic Enteritis Model Induced by Co-infection with Eimeria maxima and Clostridium perfringens[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 4120-4128.

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分数Score	病变Lesion	病变数量Number of lesions
0	无明显病变 No gross lesions	—
1	薄壁或易碎壁，或弥漫性浅表但可去除的纤维蛋白 Thin or friable walls, or diffuse superficial but removable fibrin	—
2	局灶性坏死或溃疡，或不可去除的纤维蛋白沉积 Focal necrosis or ulceration, or non-removable fibrin deposit	1~5个病灶 1 to 5 foci
3	局灶性坏死或溃疡，或不可去除的纤维蛋白沉积 Focal necrosis or ulceration, or non-removable fibrin deposit	6~15个病灶 6 to 15 foci
4	局灶性坏死或溃疡，或不可去除的纤维蛋白沉积 Focal necrosis or ulceration, or non-removable fibrin deposit	16个或更多病灶 16 or more foci
5	坏死斑块2~3 cm长Patches of necrosis 2 to 3 cm long	数量可变 Variable
6	弥漫性坏死典型的现场病例 Diffuse necrosis typical of field cases	数量可变，但广泛 Variable, but extensive

组别 Group	死亡数量 Death count	死亡时间 Time of death	存活率/% Survival rate
Control	0	—	100
CP	0	—	100
EM1	0	—	100
EM2	0	—	100
EM3	1	DPI 6	95
EM4	1	DPI 5	95
EM/CP1	0	—	100
EM/CP2	3	DPI 6	85
EM/CP3	4	DPI 5、6	80
EM/CP4	6	DPI 5、6	70

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巨型艾美耳球虫与产气荚膜梭菌共感染致鸡坏死性肠炎模型的评价

Evaluation of a Necrotic Enteritis Model Induced by Co-infection with Eimeria maxima and Clostridium perfringens

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