Ⅱd亚型微小隐孢子虫感染昆明小鼠模型的初步建立  

Preliminary establishment of Kunming mice model infected with Cryptosporidium parvumⅡd subgenotypes

作　　者：张孝天 侯文燕 张营营 张龙现[1,2] 张素梅 ZHANG Xiao-tian;HOU Wen-yan;ZHANG Ying-ying;ZHANG Long-xian;ZHANG Su-mei(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China;Key Laboratory of Quality and Safety Control of Poultry Products(Zhengzhou),Ministry of Agriculture and Rural Affairs,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物医学院,河南郑州450046 [2]农业农村部禽类产品质量安全控制重点实验室,河南郑州450046

出　　处：《中国预防兽医学报》2023年第10期1019-1024,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金面上项目(32172882、U2004155)。

摘　　要：为建立稳定、可靠的Ⅱd亚型微小隐孢子虫感染的小鼠模型,本研究以1×10^(6)个/只剂量的Ⅱd亚型微小隐孢子虫卵囊感染3周龄雌性昆明小鼠,对感染前后小鼠粪便样品进行显微镜观察和隐孢子18S r RNA和GP60基因的套式PCR检测、以及对小鼠的排卵囊规律、临床症状变化、体质量变化、回肠组织病变特征和回肠绒毛指数进行分析。结果显示:通过形态学和PCR方法均能在粪便样品中检测到微小隐孢子虫;感染组小鼠较对照组在排卵囊高峰期体质量的增加略微减少,但总体增加趋势一致,在排卵囊高峰期时感染组小鼠饮水量减少、反应迟钝、粪便变稀,高峰期后小鼠精神状态逐渐恢复良好;通过对感染后7 d小鼠的回肠组织制备病理切片并经HE染色后观察发现,与对照组相比,感染组小鼠回肠组织中出现微小隐孢子虫附着,部分区域杯状细胞数量减少,上皮细胞排列轻微紊乱且部分脱落,黏膜层可见少量炎症细胞;回肠绒毛长度显著变短、变钝,隐窝深度变浅,但绒毛直径和黏膜厚度未见显著变化。上述结果与动物临床感染隐孢子虫的症状、病理变化等基本一致,表明本研究首次建立了Ⅱd亚型微小隐孢子虫感染昆明小鼠的模型。本研究该模型的建立为研究微小隐孢子虫的致病性、宿主的免疫调节机制以及药物和疫苗的筛选提供研究基础。In order to establish a stable and reliable mouse model of Cryptosporidium parvum,1×10^(6) Cryptosporidium parvumⅡd subgenotypes oocysts were used to infect 3-week-old female Kunming mice.The feces of mice before and after infection were analyzed.The samples were subjected to microscopic observation and PCR detection.The regular pattern of oocyst shedding,changes in clinical symptoms,body weight,ileal tissue pathological characteristics and ileal villus index were analyzed.The results showed that Cryptosporidium parvum oocysts could be detected in feces through both morphology and molecular analysis;the weight gain of the infected group was slightly less than that of the control group during the peak period of ovulation sacs,but the overall increase trend was consistent.During the peak of oocyst shedding,the water consumption of mice decreased,the reaction were unresponsive,and their feces became thinner.After the peak period,the mental state of the mice gradually recovered.HE staining of pathological sections of the the ileum tissue of mice on the 7th day post-infection revealed that,compared with the control group,Cryptosporidium parvum was present in the ileum tissue of the infected mice,and the number of goblet cells was reduced in some areas.The arrangemant of epithelial cells were slightly disordered,epithelial cells were partially shed,and a small number of inflammatory cells were seen in the mucosal layer;the length of ileal villi became significantly shorter and blunted,and the depth of crypt became shallower,but no significant changes were found in villus diameter and mucosal thickness.The above results are basically consistent with the clinical infection symptoms and pathological changes in animals,indicating that this study has established a Kunming mouse model infected by Cryptosporidium parvumⅡd subgenotypes for the first time.This model provides a research basis for studying the pathogenicity of Cryptosporidium parvum,the immune regulation mechanism of the host,and drugs and vaccines screeenin

关 键 词：Ⅱd亚型微小隐孢子虫 昆明小鼠 感染模型 

分 类 号：S852.74[农业科学—基础兽医学]