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Protocol for Plasmodium falciparum Infections in Mosquitoes and Infection Phenotype Determination

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STUDIES ON MALARIAL PARASITES : VII. METHODS AND TECHNIQUES FOR CULTIVATION.

Q M Geiman¹, C B Anfinsen, R W McKee

¹Department of Comparative Pathology and Tropical Medicine, Harvard School of Public Health and Harvard Medical School, and the Department of Biological Chemistry, Harvard Medical School, Boston.

The Journal of Experimental Medicine

|October 30, 2009

Summary

Researchers developed methods to isolate Plasmodium knowlesi from monkeys for malaria studies. These techniques enable in vitro cultivation and biochemical analysis of the malaria parasite.

Keywords:

MALARIA/experimental

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Area of Science:

Parasitology
Biochemistry
Cell Biology

Background:

Plasmodium knowlesi is a malaria parasite that infects monkeys and humans.
Biochemical studies and in vitro cultivation of P. knowlesi are crucial for understanding malaria.
Efficient methods for isolating and culturing P. knowlesi are needed.

Purpose of the Study:

To describe methods for recovering adequate amounts of Plasmodium knowlesi from Macaca mulatta monkeys.
To detail techniques for the in vitro cultivation of malarial parasites.
To provide a basis for biochemical studies, drug efficacy testing, and nutritional analysis of P. knowlesi.

Main Methods:

Differential sedimentation of parasitized blood to concentrate red blood cells infected with P. knowlesi.
Rocker-dilution and rocker-perfusion methods for in vitro cultivation of malarial parasites.
Detailed descriptions of apparatus, assembly, sterilization, and parasite counting/evaluation.

Main Results:

Concentrates of P. knowlesi-infected red blood cells were obtained through differential sedimentation.
Two distinct in vitro cultivation techniques (rocker-dilution and rocker-perfusion) were established.
Control experiments showed an average multiplication rate of 3.9 over 20–24 hours.

Conclusions:

The described methods facilitate the recovery and cultivation of Plasmodium knowlesi.
These techniques are valuable for studying parasite metabolism, nutrition, and antimalarial drug action.
The findings support further research into malaria pathogenesis and treatment.