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Studies on Plasmodium ookinetes. 1. Isolation and concentration from mosquito midguts
The Journal of Protozoology
|November 1, 1976
Summary
Researchers developed a method to isolate Plasmodium berghei ookinetes from mosquito midguts using enzymatic digestion and gradient concentration. This technique yields viable ookinetes for structural and biochemical studies.
Area of Science:
- Malariology
- Parasitology
- Entomology
Background:
- Plasmodium berghei is a malaria parasite that infects mosquitoes.
- Studying Plasmodium berghei ookinetes is crucial for understanding malaria transmission.
- Previous methods for isolating ookinetes were insufficient for detailed study.
Purpose of the Study:
- To develop a reliable method for isolating concentrated Plasmodium berghei ookinetes.
- To assess the viability and purity of isolated ookinetes.
- To enable further research into ookinete structure, biochemistry, and physiology.
Main Methods:
- Isolation of ookinetes from infected Anopheles stephensi midguts.
- Enzymatic digestion to free ookinetes.
- Concentration using a BSA/Renografin gradient.
Main Results:
- A mean of 152 Plasmodium berghei ookinetes were recovered per midgut.
- Over 95% of recovered ookinetes demonstrated viability.
- Trypan blue exclusion was found to be an unreliable viability criterion.
Conclusions:
- The developed method successfully isolates viable Plasmodium berghei ookinetes.
- Contaminating microorganisms limit in vitro and in vivo development studies.
- The method allows for future research on ookinete structure, biochemistry, and physiology.