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PloS one2015; 10(4); e0125276; doi: 10.1371/journal.pone.0125276

Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites.

Abstract: 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.
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Publication Date: 2015-04-27 PubMed ID: 25915529PubMed Central: PMC4411034DOI: 10.1371/journal.pone.0125276Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses the optimization of a fluorescence-based assay for large-scale drug screening against parasites known as Babesia and Theileria. The study explores the effectiveness of anti-parasitic compounds, with the goal of discovering new treatments for these parasites.

Research Objectives

  • The primary goal of this study is to evaluate the usefulness of a fluorescence-based assay in determining the effectiveness of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures.
  • The researchers sought to determine which hematocrit (HCT, a measure of the proportion of blood volume that is occupied by red blood cells) levels—2.5%, 5%, or 10%—would yield the best results without requiring daily replacement of the medium.

Research Methodology

  • The study utilized a high-throughput screening assay (a method of testing a large number of samples rapidly) to determine the best HCT levels for different parasite species.
  • The researchers compared IC50 (half maximal inhibitory concentration, a measure of the effectiveness of a substance in inhibiting a specific biological or biochemical function) values obtained by fluorescence and by microscopy.

Research Findings

  • According to the high-throughput screening assay, the best HCT was found to be 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites.
  • The IC50 values for diminazene aceturate, luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not significantly differ whether calculated by fluorescence or microscopy.

Conclusion

  • The findings of this study demonstrate that the fluorescence-based assay, using low HCT and without requiring daily medium replacement, is an effective tool for conducting large-scale in vitro drug screening against Babesia and Theileria parasites.
  • This could pave the way for the discovery of novel chemotherapeutic agents for treating these parasites in cattle and horses.

Cite This Article

APA
Rizk MA, El-Sayed SA, Terkawi MA, Youssef MA, El Said el Sel S, Elsayed G, El-Khodery S, El-Ashker M, Elsify A, Omar M, Salama A, Yokoyama N, Igarashi I. (2015). Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites. PLoS One, 10(4), e0125276. https://doi.org/10.1371/journal.pone.0125276

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 10
Issue: 4
Pages: e0125276
PII: e0125276

Researcher Affiliations

Rizk, Mohamed Abdo
  • 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.
El-Sayed, Shimaa Abd El-Salam
  • 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.
Terkawi, Mohamed Alaa
  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido, Japan.
Youssef, Mohamed Ahmed
  • Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
El Said, El Said El Shirbini
  • Department of Biochemistry and Chemistry of Nutrition, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
Elsayed, Gehad
  • Department of Biochemistry and Chemistry of Nutrition, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
El-Khodery, Sabry
  • Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
El-Ashker, Maged
  • Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
Elsify, Ahmed
  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido, Japan; Department of Animal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Minoufiya University, Sadat City, Minoufiya, Egypt.
Omar, Mosaab
  • Department of Parasitology, Faculty of Veterinary Medicine, South Valley University, Qena, Egypt.
Salama, Akram
  • Department of Animal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Minoufiya University, Sadat City, Minoufiya, Egypt.
Yokoyama, Naoaki
  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido, Japan.
Igarashi, Ikuo
  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido, Japan.

MeSH Terms

  • Animals
  • Antiparasitic Agents / chemistry
  • Antiparasitic Agents / pharmacology
  • Babesia / drug effects
  • Babesia / growth & development
  • Cattle / parasitology
  • Cells, Cultured
  • Drug Evaluation, Preclinical / methods
  • Hematocrit
  • Horses / parasitology
  • Microscopy, Fluorescence
  • Theileria / drug effects
  • Theileria / growth & development

Conflict of Interest Statement

Ikuo Igarashi is a PLOS ONE Editorial Board member, however the review, processing and decision about this paper will not alter the authors' adherence to PLOS ONE editorial policies and criteria.

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