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柔嫩艾美耳球虫(球虫纲)子孢子对离子载体抗球虫作用反应的流式细胞术分析

Flow cytometric analysis of the response of Eimeria tenella (Coccidia) sporozoites to coccidiocidal effects of ionophores.

作者信息

Fuller A L, Golden J, McDougald L R

机构信息

Department of Poultry Science, University of Georgia, Athens 30602, USA.

出版信息

J Parasitol. 1995 Dec;81(6):985-8.

PMID:8544076
Abstract

Fluorescein diacetate (FDA) and propidium iodide (PI) were used as indicators of membrane integrity after Eimeria tenella sporozoites were treated with polyether ionophores. Flow cytometry was used to quantitate the structural and functional effects based on red or green fluorescence and shape index of the sporozoites. Two field isolates (FS119 and FS139) were essentially resistant to polyether ionophores administered under practical conditions, whereas a laboratory strain was considered sensitive. The shape of sporozoites changed after treatment with ionophores, and this could be detected by flow cytometry. Green-fluorescing cells declined in number as the membranes were compromised by ionophore treatment. Red-fluorescing cells increased as the compromised membranes allowed entry of PI to bind with the nucleic acids. These effects were generally slower to develop in ionophore-tolerant field isolates compared with the sensitive laboratory strain. The effect of lasalocid on FDA and PI uptake, change in shape of the sporozoites, and lysis of sporozoites was more rapid than that of monensin or salinomycin. The 2 field isolates responded at different rates to salinomycin and monensin. Flow cytometry was a sensitive and accurate instrument for analysis of the effects of ionophores on sensitive and resistant lines of coccidia.

摘要

在用聚醚离子载体处理柔嫩艾美耳球虫子孢子后,使用二乙酸荧光素(FDA)和碘化丙啶(PI)作为膜完整性的指标。流式细胞术用于根据子孢子的红色或绿色荧光以及形状指数来定量其结构和功能效应。在实际条件下,两个田间分离株(FS119和FS139)对施用的聚醚离子载体基本耐药,而一个实验室菌株被认为敏感。用离子载体处理后子孢子的形状发生了变化,这可以通过流式细胞术检测到。随着离子载体处理使膜受损,发绿色荧光的细胞数量减少。随着受损的膜允许PI进入与核酸结合,发红色荧光的细胞增加。与敏感的实验室菌株相比,这些效应在耐离子载体的田间分离株中通常发展得较慢。拉沙洛西对FDA和PI摄取、子孢子形状变化以及子孢子裂解的影响比莫能菌素或盐霉素更快。这两个田间分离株对盐霉素和莫能菌素的反应速率不同。流式细胞术是分析离子载体对球虫敏感和耐药品系影响的灵敏且准确的仪器。

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