摘要: 目的建立兔斯氏艾美耳球虫(Eimeria stiedai,E.stiedai)感染模型和巢式PCR诊断方法。方法利用剖检、显微镜观察、血液生化和病理组织学检查等方法对建立的兔E.stiedai模型进行鉴定。通过收集卵囊,提取E.stiedai的DNA,设计特异性引物,建立E.stiedai巢式PCR检测方法。结果临床剖检:兔腹围明显增大,解剖见肝脏肿大,肝脏表面和实质布满白色及淡黄色结节,胆囊和胆管肿大,胆汁呈淡黄色。显微镜观察:虫卵大小为(31.72～38.43)μm×(18.10～22.69)μm。血液生化检查:球蛋白(GLOB)指标偏高,肌酐(CREA)和碱性磷酸酶(ALKP)指标偏低,其余检测指标均在参考值范围内。病理组织学检查:肝组织和胆管中见大量粉红色E.stiedai虫卵。巢式PCR检查:最低检测限为1个卵囊DNA样本和1.14×103拷贝数质粒,阴性对照和空白对照均未出现条带,重复性实验变异性系数<5%。结论成功构建兔E.stiedai感染模型,建立的巢式PCR诊断方法可扩增出E. stiedai的特异片段,敏感性强,重复性好。

摘要: The experimental beastie in this essay is only the rodent (rat, small rat, shrewmouse, cavy) and rabbit. The eimeria parasitizing in beastie are variety and frequency. Some of them are distinctly pathogeny and dangerous, so eimeria is one of debared parasites in “International Standards". All the eimerias in beastie are not touched in our country except rabbit eimeria reported systematically, so we aim to discuss it in this essay. The following are our opinions based on the information we have. 1. The 30 merias can be divided into 5 animal groups according to the eimeria's special advocation characteristic. 2. The classification nowadays is based on the conformation characteristic. It is difficult to distinguish when there are many kinds of merias, because the maturate characteristics of oocyst are different. So it is better to see if the sample is new or old when you choose the material. You can find the rule by contrasting, so that the distinguish of species is easier. 3. The conformation of oocyst is the main point of classification. We can increase the creditability of defination by the colligate judgement of observing the transformation in organize pathology cooperating the entirely process of eimeria life history, such as the asexuality procreation and sexual reproduction of eimeria in intestines cell and the form of oocyst.

摘要: 目的探求一种安全高效的驱治高原鼠兔球虫的方法。方法选用复方伊维菌素干悬浮剂和天字球虫粉2种抗寄生虫药,采取经口给药(灌服和加入饲料、饮水),观察驱治效果。结果实验室检测到高原鼠兔感染的球虫属艾美尔科,艾美尔属,初步鉴定出5个种,总感染率87.09%(108/124)。给药1 d后,阳性率下降为39.51%,30 d后为14.51%,第45 d为0。结论针对野生感染的球虫,采用复方伊维菌素干悬浮剂和天字球虫粉联合驱治效果良好,为人工饲养高原鼠兔的寄生虫病防治和寄生虫质量监测提供参考。

摘要: 目的 研究堆型艾美尔球虫延伸因子1-α(EaEF-1α)在堆型艾美尔球虫的感染期间调控细胞凋亡的主要机制。方法 从球虫裂殖子侵染DF-1细胞实验的上清液中分析筛选得到EaEF-1α,与质粒pEGFP-N1重组得到重组质粒pEGFP-N1-EaEF-1α,转染入DF-1细胞中，Western bolt及荧光检测蛋白表达，通过流式细胞术检测脂多糖组、pEGFP-N1组、pEGFP-N1-EF-1α组和空白组DF-1细胞凋亡情况，酶联免疫吸附实验检测脂多糖组、pEGFP-N1组、pEGFP-N1-EF-1α组和空白组DF-1细胞中凋亡因子Bax、Bcl-2、Caspase3、Caspase8表达水平。结果 流式细胞术检测发现pEGFP-N1-EF-1α组细胞凋亡率显著低于pEGFP-N1组和脂多糖组。DF-1细胞裂解液中Bax表达水平降低，Bcl-2及Caspase3表达水平上调，pEGFP-N1-EF-1α组Bcl-2/Bax的值显著高于pEGFP-N1组和脂多糖组。Caspase8表达水平无差异。结论 EaEF-1α体外过表达通过线粒体通路显著抑制细胞凋亡。