An in vitro larval migration assay for assessing anthelmintic activity of different drug classes against Ascaris suum.
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 (EC50). The values of EC50of 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.
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.
Jianguo
J
Zhao
Department of Veterinary Disease Biology, University of Copenhagen, DK-1870, Frederiksberg C, Denmark.
Andrew R
AR
Williams
Department of Veterinary Disease Biology, University of Copenhagen, DK-1870, Frederiksberg C, Denmark.
Tina Vicky Alstrup
TVA
Hansen
Department of Veterinary Disease Biology, University of Copenhagen, DK-1870, Frederiksberg C, Denmark.
Stig M
SM
Thamsborg
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, 730046, China.
Jianping
J
Cai
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.
Shuaibao
S
Song
College of Agriculture and Animal Science, Qinghai University, Xining, Qinghai 810016, China.
Gang
G
Chen
College of Agriculture and Animal Science, Qinghai University, Xining, Qinghai 810016, China.
Ming
M
Kang
Xinjiang Veterinary Research Institute, Xinjiang Academy of Animal Science, Urumqi, Xinjiang 830000, China.
Zhuangzhi
Z
Zhang
National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
Qun
Q
Liu
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. Electronic address: qianhan@hainu.edu.cn.
Qian
Q
Han
D000362:Agar; D000818:Animals; D000871:Anthelmintics / Q000145:classification / Q000494:pharmacology*; D017165:Ascaris suum / Q000187:drug effects* / Q000502:physiology; D001681:Biological Assay / Q000379:methods*; D007814:Larva / Q000187:drug effects / Q000502:physiology; D009043:Motor Activity / Q000187:drug effects*
D000871:Anthelmintics; D000362:Agar
Agar; Anthelmintic; Ascaris suum; Benzimidazole; Larval migration assay; Parasitic nematode