青海部分地区藏羊膈肌住肉孢子虫感染情况调查及分子生物学种属鉴定

发布时间：2018-03-19 16:22

本文选题：藏羊　切入点：住肉孢子虫　出处：《青海大学》2015年硕士论文　论文类型：学位论文

【摘要】：采用压片镜检法对青海省黄南州泽库县、海北州祁连县和海晏县、海西州乌兰县、海南州共和县和贵南县、海东市互助县和化隆县共八个县的181只藏羊膈肌住肉孢子虫感染情况进行了调查。结果表明,共检出阳性藏羊141只,其中感染率最高为乌兰县、互助县(100%),感染强度最高为乌兰县,达368(14~1296)个/g。显微镜下测得八县虫体大小的平均值为0.35(0.15~1.20)mm×0.05(0.04~0.13)mm。调查结果提示,青海省部分地区藏羊膈肌住肉孢子虫的感染情况较为严重,其防治工作应引起相关部门足够的重视。对检出的阳性膈肌,经切片制作、HE染色后进行了虫体包囊形态结构的观察,结果表明:藏羊膈肌住肉孢子虫虫体包囊呈梭形,少数呈不规则形状;包囊寄生部位的肌纤维明显肿胀,肌组织结构模糊;虫体包囊壁未见特殊形态;扫描电镜观察了虫体包囊的超微结构,结果显示:虫体纵切呈梭形,横切呈豌豆形;囊壁有原生囊壁和次生囊壁组成,厚约5um;原生囊壁光滑,次生囊壁较为粗糙,基质向囊腔内延伸,将囊腔分隔成若干个不同的小室,小室内含有香蕉形的缓殖子。以18S r RNA作为靶基因,对寄生于藏羊膈肌的住肉孢子虫的目的基因进行特异性扩增、测序、构建了系统发育树。分析表明:所测样品基因序列与Gen Bank上已公布的柔嫩住肉孢子虫的基因序列高度相似。综合染色切片形态观察、扫描电镜虫体包囊形态、18S r RNA基因测序结果,将采自青海藏羊膈肌的住肉孢子虫鉴定为羊柔嫩住肉孢子虫(Sarcocystis tenella)。
[Abstract]:The methods of pressing microscopic examination were used to examine Zecu County of Huangnan Prefecture, Qilian County and Haiyan County of Haibei Prefecture, Wulan County of Haixi Prefecture, Republican County of Hainan State and Guinan County of Qinghai Province. An investigation was carried out on 181 Tibetan sheep infected with the diaphragm in eight counties of Haidong and Hualong. The results showed that 141 Tibetan sheep were positive, and the highest infection rate was in Wulan County. The infection intensity was the highest in Wulan county (368,141,296) / g. The average size of insect body measured under microscope was 0.35 ~ 0.15 ~ 1.20mm 脳 0.05 脳 0.05 ~ 0.04 ~ (0.13) mm. the results of investigation indicated that the infection of diaphragm of Tibetan sheep was more serious in Qinghai province than that in other parts of Qinghai Province, and the results showed that the average size of insect body was 0.35 ~ 0.15 ~ 1.20 mm 脳 0.05 ~ 0. 05 ~ (0. 05) ~ 0. 13 m ~ (m ~ (m)). The prevention and cure work should be paid enough attention to by relevant departments. The morphology and structure of the cysts of the positive diaphragm detected by HE staining were observed. The results showed that the cysts of the diaphragm of Tibetan sheep were fusiform. A few of them were irregular in shape; the muscle fibers in the parasitic part of the cyst were obviously swollen and the structure of the muscle tissue was blurred; the wall of the cyst did not show a special shape; the ultrastructure of the cyst was observed by scanning electron microscope. The results showed that the longitudinal cutting of the insect body was fusiform. The primary wall is smooth, the secondary wall is rough, and the matrix extends into the cavity, dividing the cavity into several different chambers, which are composed of primary and secondary walls of the capsule, which are in the shape of peas, and have a thickness of about 5 um.The primary wall is smooth, the secondary wall is rough, and the matrix extends into the cavity of the capsule. Using 18s r RNA as target gene, the specific amplification and sequencing of the target gene of Sarcocystis infestans were carried out in the diaphragm of Tibetan sheep. Phylogenetic tree was constructed. The results showed that the gene sequences of the samples were highly similar to those of Gen Bank. The results of scanning electron microscopy (SEM) showed that the sarcocystis tenellae was identified as Sarcocystis tenellaa, which was collected from the diaphragm of Tibetan sheep in Qinghai, and was identified as Sarcocystis tenellaa.
【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.26

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