粪便保存条件、时间对鸡球虫卵囊活力的影响实验

为探讨在不同温度条件下粪便保存不同时间对球虫卵囊成熟力的影响,本实验将含有球虫卵囊的粪便在-20℃、4℃、20℃、28℃分别保存0 h、24 h、48 h、72 h,通过观察卵囊的孢子化率确定最佳保存条件。结果表明:相同保存温度条件下,保存0 h、24 h、48 h、72 h的E.tenella早熟耐药株卵囊的孢子化率依次降低。-20℃和28℃时孢子化率均未达到80%;在4℃时粪便保存24 h的孢子化率达到80%以上,保存48h及以上时孢子化率极显著(P0.01)低于保存24 h的孢子化率;在20℃时的孢子化率均达到80%以上。     

鸡球虫亚单位苗研究进展

目前药物治疗和活菌预防对球虫病来说存在许多问题，所以研究者的工作重点主要转移到重组亚单位苗的开发上，并且已经克隆出许多抗原。本文从母源亚单位苗和重组亚单位苗两方面对球虫进行了阐述。特别是对柔嫩艾美耳球虫和堆型艾美耳球虫的重组抗原的研究现状进行了较为详细的综述。     

鸡艾美耳球虫3-1E基因的克隆与表达

应用反转录 -聚合酶链式反应 (RT- PCR)技术 ,分别从柔嫩艾美耳球虫甘肃株 (E.tenella GS,Et GS)和堆型艾美耳球虫青海株 (E.acervulina QH,Ea QH)孢子化卵囊子孢子中提取的总 RNA扩增得到鸡球虫子孢子表面抗原 3- 1E基因 (Et GS3-1E和 Ea QH 3- 1E)。将 Et GS3- 1E与原核表达载体 p GEX- 6 P1连接 ,构建了的 p GEX- 3- 1E原核表达质粒 ,并获得融合蛋白的高效表达和纯化。序列分析表明 :Et GS3- 1E和 Ea QH 3- 1E的 ORF均为 5 13个碱基 ,共编码 171个氨基酸。Et GS3- 1E的蛋白分子量为 18.5 k D,Ea QH 3- 1E的蛋白分子量为 18.6 k D。ORF比较 ,Et GS3- 1E与文献报道的 Ea3- 1E比较 ,共有 2个核苷酸发生变异 ,核苷酸同源性为 98.8%,有 1个氨基酸发生变异 ,氨基酸同源性为 99.4 %;Ea QH 3- 1E与文献报道的 Ea3- 1E比较 ,共有 4个核苷酸发生变异 ,核苷酸同源性为 97.7%,有 3个氨基酸变异 ,氨基酸同源性为 98.3%;Et GS 3- 1E与 Ea QH 3- 1E比较 ,有 3个核苷酸发生变异 ,核苷酸同源性为 98.2 %,有 2个氨基酸发生变异 ,氨基酸同源性为 98.8%。获得了 Et GS 3- 1E融合蛋白的高效表达和纯化 ,表达率达 4 3.2 %。     

Ultrastructure of Goussia cruciata (Apicomplexa: Coccidia) infecting the liver of horse mackerel, Trachurus trachurus (L.), from Ibero-Atlantic waters

The ultrastructure of developmental stages of Goussia cruciata and the pathology they cause in the liver of Trachurus trachurus (Teleostei: Carangidae) caught off the Galician (North-West Spain) and Portuguese North Atlantic coasts are described. Each oocyst contained four ellipsoidal sporocysts, with two sporozoites. The sporocyst wall consisted of a thick and dense inner layer with transverse striations and a multi-lamellated outer layer formed by parallel dense internal bands alternating with lighter areas. The lamellae formed filamentous extensions of the wall. The sporocyst wall striation period was smaller than that observed in G. clupearum, which has a similar habitat. The dehiscence suture, characteristic of the genus, was present in the sporocysts of G. cruciata. The sporocysts were arranged in a symmetrical and characteristic cross shape. A large number of sporocysts with sporozoites were observed in direct contact with host liver cells. No macroscopic lesions were observed. In heavily infected fish, aggregations of oocysts were often enveloped in a 'yellow body' composed of amylopectin granules derived from the parasite and necrotic or aggregated host cells. Degenerating parasites were frequently observed in liver tissue. Host inflammatory cells were accumulated near some oocysts. The ultrastructure of the parasite, together with its strict host specificity, confirmed G. cruciata as a separate and valid species.