Anti-plasmodial Property Of Moringa Oleifera Seed Extract On Swiss Mice

3.5 Experimental design
The animals were divided into five (5) groups consisting of four (4) animals each. The malaria parasite density of the experimental animals in each group was determined before the commencement of the administration of the seed extract. Each of the groups is described as follows:
Group 1 (Test group): The animals in this group were administered with Moringa oleifera seed extract at a concentration of forty percent (40%) orally twice daily at interval of 12hrs.
Group 2 (Test group): The animals in this group were administered with Moringa oleifera seed extract at a concentration of sixty percent (60%) orally twice daily at interval of 12hrs.
Group 3(Test group): The animals in this group were administered with Moringa oleifera seed extract at a concentration of eighty percent (80%) orally twice daily at interval of 12hrs.
Group 4 (Test group): The animals in this group were administered with Moringa oleifera seed extract at a concentration of one hundred percent (100%) orally twice daily at interval of 12hrs.
Group 5 (Negative Control): The animals in this group consist of four (4) were administered with chloroquine orally twice daily at interval of 12hrs .
Group 6(Positive Control): the animals in this group consist of four (4) swiss mice and were placed on Placebo by administering distill water twice daily also at interval of 12hrs
3.6 Collection and inoculation of Malaria Parasite Strain
The rodent malaria parasite, Plasmodium berghei NK 65 strain was obtained from a donor
(infected) mouse at the Instituteof Advanced Medical Research and Training (IMRAT), University of Ibadan, Ibadan. Plasmodium. berghei-infected red blood cells was obtained from the tail vein of infected mouse by cuttingthe tip of its tail with a sterile scissors, this was then diluted with phosphate buffered saline(PBS) so that each 0.2 ml that was subsequently injected contained approximately 107 infected red cells (> 40% parasitemia) per kilogram of body weight.
    The parasite preparation (containing P. berghei) was injected intraperitoneally into 32healthy Swiss mice using a sterile syringe as described by David et al., (2004). After three days, the pre-intervention test was carried out to confirm the presence of the parasite. This was done by collecting each mouse’s blood on separate slides. The blood was obtained by cutting the tip of each mouse tail with a sterile scissors and massaging it gently to draw blood. Each blood sample was collected in duplicates on two clean slides, allowed to dry and stained by Leishman technique as described by Chessbrough (2000).  All stained slides were examined under x100 objective of the light microscope to determine the percentage of parasitemia. Only concordant results were recorded while discordant ones repeated. Percentage suppression / inhibition was also calculated after four days of extract administration. The formulae below were used for the calculations:

3.7 Statistical analysis
Results obtained were analyzed using the SPSS version 21 software (SPSS Inc, USA). The data were expressed as mean ±Standard error of mean (SEM). The test of Significance was performed using student t–test for analysis of data from the test and control groups of the experimental animals. The level of significance was based on P value less than 0.05.