一种用于评估不同药物类别对猪蛔虫驱虫活性的体外幼虫迁移试验。

An in vitro larval migration assay for assessing anthelmintic activity of different drug classes against Ascaris suum.

作者信息

Zhao Jianguo, Williams Andrew R, Hansen Tina Vicky Alstrup, Thamsborg Stig M, Cai Jianping, Song Shuaibao, Chen Gang, Kang Ming, Zhang Zhuangzhi, Liu Qun, Han Qian

机构信息

Laboratory of Tropical Veterinary Medicine and Vector Biology, and Hainan Key Laboratory of Sustainable Utilization of Tropical Bioresources, College of Agriculture, Hainan University, Haikou 570228, China; Key Laboratory of Tropical Biological Resources of Ministry of Education, Hainan University, Haikou, Hainan 570228, China; State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, 730046, China.

Department of Veterinary Disease Biology, University of Copenhagen, DK-1870, Frederiksberg C, Denmark.

出版信息

Vet Parasitol. 2017 Apr 30;238:43-48. doi: 10.1016/j.vetpar.2017.03.014. Epub 2017 Mar 18.

DOI:10.1016/j.vetpar.2017.03.014

PMID:28342672

Abstract

In vitro methods have been developed for the detection of anthelmintic resistance in a range of nematode species. However, the life cycle of Ascaris suum renders the commonly used egg hatch assay and larval development assay unusable. In this study we developed a combined multi-well culture and agar gel larval migration assay to test the effect of benzimidazole and tetrahydropyrimidin/imidazothiazole anthelmintics against nine isolates of A. suum collected from locations in China and Denmark. Drugs tested were thiabendazole, fenbendazole, mebendazole, levamisole, and pyrantel. The percentages of larvae that migrated to the surface of each treated and control well were used to calculate the drug concentration which inhibits 50% of the larvae migration (EC). The values of EC of thiabendazole, fenbendazole, mebendazole, levamisole, and pyrantel against A. suum isolates ranged 74-150, 4.9-13.9, 2.3-4.3, 358-1150 and 1100-4000nM, respectively. This combined multi-well culture and agar gel larval migration assay was a sensitive bioassay for anthelmintic activity and could serve as an in vitro method to detect for lowered drug efficacy against A. suum or possibly to screen for anthelmintic drug candidates.

摘要

已开发出体外方法用于检测一系列线虫物种中的抗蠕虫药耐药性。然而，猪蛔虫的生命周期使得常用的虫卵孵化试验和幼虫发育试验无法使用。在本研究中，我们开发了一种组合式多孔培养和琼脂凝胶幼虫迁移试验，以测试苯并咪唑和四氢嘧啶/咪唑噻唑类抗蠕虫药对从中国和丹麦各地收集的9株猪蛔虫分离株的作用。所测试的药物有噻苯达唑、芬苯达唑、甲苯达唑、左旋咪唑和噻嘧啶。用迁移到每个处理孔和对照孔表面的幼虫百分比来计算抑制50%幼虫迁移的药物浓度（EC）。噻苯达唑、芬苯达唑、甲苯达唑、左旋咪唑和噻嘧啶对猪蛔虫分离株的EC值分别为74 - 150、4.9 - 13.9、2.3 - 4.3、358 - 1150和1100 - 4000 nM。这种组合式多孔培养和琼脂凝胶幼虫迁移试验是一种用于检测抗蠕虫药活性的灵敏生物测定法，可作为一种体外方法来检测针对猪蛔虫的药物疗效降低情况，或可能用于筛选抗蠕虫药候选物。