Effects of Heparin, Acid Citrate Dextrose, and EDTA on Plasmodium Falciparum Parasites Morphology and in Vitro Growth Rate

Abstract

This study investigated the effects of three anticoagulants - Acid Citrate Dextrose (ACD), Heparin, and EDTA - on the in vitro growth and morphology of Plasmodium falciparum (3D7 strain). Using venous blood from five healthy donors, we established triplicate cultures for each anticoagulant and monitored parasitaemia daily for 14 days through microscopic examination. Our results demonstrated distinct patterns of parasite growth across the different anticoagulant conditions. ACD-treated cultures showed the most favorable growth characteristics, reaching peak parasitaemia of 13.57% and maintaining well-preserved parasite morphology throughout the observation period. Heparin-supported cultures exhibited moderate growth (peak 7.4% parasitaemia) with some morphological variations, while EDTA-treated cultures displayed the lowest growth rates (peak 4.13% parasitaemia) and more frequent degenerative changes in parasite forms. Statistical analysis using ANOVA (F = 0.404, p > 0.05) revealed no significant differences in mean parasitaemia levels among the three anticoagulant groups. However, the consistent trend of superior performance in ACD-treated cultures suggests its potential advantage for maintaining robust P. falciparum cultures. These findings have important implications for malaria research methodologies, particularly in studies requiring optimal parasite viability and standardized growth conditions. The study provides evidence-based recommendations for anticoagulant selection in P. falciparum culture systems, with ACD emerging as the preferred choice for high-yield cultures, while Heparin and EDTA may serve as viable alternatives depending on specific experimental requirementsThese results contribute to the standardization of malaria research protocols and may enhance the reliability of in vitro studies investigating antimalarial drug efficacy and parasite biology.