Rizk Mohamed Abdo, El-Sayed Shimaa Abd El-Salam, Terkawi Mohamed Alaa, Youssef Mohamed Ahmed, El Said El Said El Shirbini, Elsayed Gehad, El-Khodery Sabry, El-Ashker Maged, Elsify Ahmed, Omar Mosaab, Salama Akram, Yokoyama Naoaki, Igarashi Ikuo
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido, Japan; Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido, Japan; Department of Biochemistry and Chemistry of Nutrition, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
PLoS One. 2015 Apr 27;10(4):e0125276. doi: 10.1371/journal.pone.0125276. eCollection 2015.
A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses.
为发现针对巴贝斯虫寄生虫的新型化疗药物，需要一种快速准确的大规模评估抗巴贝斯虫药物的检测方法。在本研究中，我们评估了一种基于荧光的检测方法在体外培养中测定抗巴贝斯虫化合物对牛和马血液寄生虫疗效的实用性。使用了三种不同的血细胞比容（HCT；2.5%、5%和10%），且不每天更换培养基。高通量筛选检测结果显示，对于牛巴贝斯虫寄生虫，最佳HCT为2.5%，对于马巴贝斯虫和泰勒虫寄生虫，最佳HCT为5%。通过荧光法和显微镜法获得的乙酰氨基阿维菌素的IC50值没有显著差异。同样，木犀草素、磷酸咯萘啶、印楝素、格杜宁和依诺沙星的IC50值在两种方法之间也没有差异。总之，我们基于荧光的检测方法使用低血细胞比容，且不需要每天更换培养基，非常适合针对感染牛和马的巴贝斯虫和泰勒虫寄生虫进行体外大规模药物筛选。
