Adeola Aderounmu(Download word doc of full research report Adeola’s phd report)
Preamble…
The use of coconut water seems to be the only previous attempt made to cultivate P. falciparum using plant medium as reported by Renapurkar and Sutar (1989). Even then, RPMI and human serum were employed because the red cells were lysed when coconut water was used alone. However, by more thoroughly defining the nutritional requirements of P. falciparum, it may be possible to eliminate the human serum (and RPMI 1640) required for the continuous cultivation or to use animal sera without any reduction in growth (Divo and Jensen, 1982b). One of the potential uses of such a medium is in the standardization of antimalarial drug sensitivity tests (Ofulla et al., 1994).
Growing Malaria Parasites in Plant Based Medium. (A hidden recipe for malaria vaccine antigen?)
In this new attempt which was the core of my PhD research:
P. falciparum was successfully cultivated in vitro in a medium that is free of both human serum and RPMI 1640. The features of this newly developed serum free formulation require further optimization and investigation into its ability and sustainability for continuous cultivation of P. falciparum. Approaches such as this are also aimed to eliminating host factors influence in parasite biochemical and immunological experiments.
This may probably be the first successful cultivation of P. falciparum using a plant exudate at basal level of nutrients and animal extracts to sustain development. Arguably, it also represents the cheapest way to keep the parasite alive.
References:
Renapurkar, D. M and Sutar, N. K. (1989) Coconut water and the cultivation of Plasmodium in vitro. Trans R. Soc. Trop. Med. Hyg. 83, 720.
Divo, A. A and Jensen, J. B. (1982b) Studies on serum requirements for the cultivation of P. falciparum 2. Medium enrichment. Bull. W. H. O. 60, 571-575
Ofulla, A V. O., Orago, A. S., Githure J. I., Burans, J. P Aleman, G. M., Johnson, A. J and Martin, S. K. (1994) Determination of fifty percent inhibitory concentration (IC50) of antimalarial drugs against P. falciparum parasites in a serum-free medium. Am. J. Trop. Med. Hyg. 51, 214-218.
The PhD Thesis Experiments:
Study on malaria culture system
Preparation of Glucose solution
Preparation of saline solution
Preparation of (Phosphate buffer) PBS
Preparation of liver extract
Preparation of sap (plant exudate) solution
Preparation of hypoxanthine solution
Effects of water dilution of stock sap solution on uninfected erythrocytes
The newly formulated plant medium for growing malaria parasite
New Malaria culture experiment
The influence of serum variability of the in vitro cultivation of Plasmodium falciparum
Materials and methodology
Synchronization of parasite culture
The use of plant extract and human serum for the cultivation of P. falciparum malaria parasites
Studies on antimalarial plants
Plants used
Preparation of extracts
Culture technique
Fluorescent microscopy
Antimalarial crude extract combination
The plant extracts combinations
Plant extracts
In vitro inhibition assay for extract combination
Stage specificty of activity of antimalarial extracts
Materials and methods for studies on insecticides treted curtains
Study population for Insecticides treated Nets usage
Enzyme-linked immunosorbent assay (ELISA)
Preparation of parasite DNA
Polymerase chain reaction (PCR)
Gel electrophoresis
The Results
New malaria culture
Influence of serum variability
On antimalarial plants
Insecticides treated curtain
P. falciparum specific IgG Parasite specific
IgG1 and IgG3 antibodies
P.falciparum genotypes
Total IgG. IgG1 and IgG3 subclsses.
P.falciparum genotypes
The Discussions
Malaria culture system
On antimalarial plants
On insecticides treated curtains
Multiplicity of Infection
Summary
References
Click Adeola’s phd report for the full text/Full Thesis in word document.
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