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.

All questions/enquiries to aderounmu@gmail.com

Statistical analysis using the chi-square concept was used to determine the prevalence pattern of malaria in metropolitan Lagos. Data used for the analysis were collated from the parasitology laboratory of the Lagos University Teaching Hospital from 1991 to 1999. Over 6 500 blood samples were examined during the period.

The (modified) word document which appeared here was actually submitted for publication in its original form but it was rejected. I wished I never even try to publish it. All these publication stuff and rejection thing can drive you mad if you are not mentally strong as a hardworking scientist.

Here it is now, published for free!!!

incidence-of-malaria-in-lagos.doc

When the plants were combined, they were more effective.

Enantia chlorantha is a promising antimalarial plant and it was combined with each of the other 3 that were used in the first experiment.

See the word document for details.

Toxicity test were never conducted in this experiment and the active substances were not isolated.

Open word doc: combination_blog3.doc

Here I present the first part of my work with 4 medicinal plants that have antimalarial activity. These plants are commonly used in Nigeria against fever as well.

Word doc: (Might take some time to open, be patient!)

medicinal-plants-1_blog.doc

This may sound like an impossibility but it happened! I was able to sustain malaria parasite in a medium that is plant based.

Here is my research from 2002 in word document.

malaria-culture-in-plant-medium_blog.doc

My first research led to the invention of a mechanisms that allows the Malaria parasite-Plasmodium to be sustained in plant-based medium for a limited number of days. Perhaps, one can say that the parasite can grow in this medium. The medium will need further optimisations in order to be able to perform like the conventional growth media.

In other area, some of the ingredients in the medium would perhaps require artificial synthesis on the long run since the use of mice liver extract may not be very easy to comply with. (See thesis for further details).

The second aspect of my PhD research was the use of 4 medicinal plants extracts in inhibitory experiments (in vitro). The extrats used showed inhibitory properties which were more effective when combined. The treatment of malaria today depends on combination of drugs in any case. (See thesis for further details).

I did not find out the active ingredients in the plants since that was beyond the scope of my personal ability and all attempts to get collaborations in Sweden (Karolinska, Goteborg University, Uppsala Uni.) and outside Sweden failed! No one was interested in my plan of action or they said it was too advanced…

The third aspect was on Immunology of Malaria. The best way to deal with malaria will be the use of vaccine. This remains elusive. ITN does save life! (see thesis for details)

This blog is dedicated to my phd research.  

Please note that I did my PhD quite alright but I do not use PhD because I didn’t defend my thesis. I was never awarded a PhD degree by any institution in Nigeria or abroad.

The American Insitutions or whoever addresses me as Dr. Aderounmu are doing that on their own personal judgements!

All my research reports will appear on this blog. They will be released gradually…as time permits me.

Check on PAGES to the right to find out more about my research and the facilitators.

Welcome back later.

Thank you.

Adeola

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Updated: 20th Nov 2007.

My thesis is now available on this blog. There is only a little more information to be added. Just a few updates here and there.

Goodluck to all prospective malaria phd students and researchers. I hope you sincerely know what you are doing and why you are in research……