Analysis of Selected Medicinal Plants used in the Treatment of Malaria and Typhoid Fever in Ebonyi State , Nigeria

The study was aimed at evaluating selected medicinal plants used in the treatment of malaria and typhoid fever. Materials used include medicinal plants used locally in treating malaria and typhoid fever sourced from different villages in Ebonyi State. The analysis of the medicinal plants was conducted using random amplified polymorphic DNA (RAPD) markers and NTSYSpc software version 2.02. Different RAPD markers including OPB-1, OPB-2, OPB-3, OPB-5, OPB-12 and OPH-12 were used to amplify the DNA of these plants. These markers were found to be polymorphic except OPB-3 which did not produce any band. It was observed that RAPD markers can effectively amplify DNA sequences of different medicinal plants. The data matrix of RAPD profiles obtained from fragments of each amplicon were scored as 1 (presence of alleles) or 0 (absence of alleles). A dendrogram of the plants using unweighted pair group mean (UPGMA) clustered the genotypes into groups. The dissimilarity values were 0.26 and 1 as minimum and maximum with an edge length of 1.32. Principal component analysis of the generated amplicons resulted to clusters with unique genetic identity. The polymorphism detected among the plants genotypes will be useful in selecting genetically diverse species in future breeding programme. Keywords: Medicinal plants, Malaria/Typhoid, RAPD, Ebonyi State, Nigeria

Herbal medication remains the major unavailability of health post in some rural areas of the state and the cost of treatment, it is sources of healing for most persons infected with imperative to identify, authenticate and ensure malaria and typhoid fever.Because of its broad the availability of plant species with the potential knowledge structure, herbal medication involves of curing.This study analyzed selected medicinal the utilization of substances, dosage and plants in Ebonyi State used in the treatment of practices due to social and cultural norms and malaria and typhoid fever using RAPD markers.beliefs and also showcases the know-hows and thought of some of people.In order to Materials and Methods determine, avoid and cure societal and spiritual Plant samples were collected from disparity this idea is spread across generations villages in Ebonyi State and identified by a (Diallo and Paulsen, 2000).
qualified taxonomist in the Department of About 80% of the global population Applied Biology, Ebonyi State University depends on herbal treatment for their health Abakaliki, Ebonyi State.care delivery, and most of these people use plants or their ethics (Gupta et al. 2005) and iso-amylacohol at 25: 24: 1 was added.It these plant natural resources for their health was vortexed and centrifuged at 1, 3000 g for 10 care.
However, because of high price in min and 450 µl of the supernatant was removed availability, cultural mismatch and selfand poured into 1.5 ml sterile tube and 400 µl of sufficiency several persons are presently cold isopropanol was added into the resorting to folk practice for primary health supernatant.It was mixed and incubated for 3 h o at 20 C in freezer and centrifuge at 1,3000 g for treatment.(Kamatenesi-Mugisha et al.2005).
10 min to sediment the DNA.The supernatant Based on ethic favorites and apparent efficacy was decanted to ensure that the pellets was not some people also adopt traditional medicines distributed and 500 µl of 70% ethanol was added (WHO, 2002).
to the pellets and centrifuged again at 1,3000 g The physiological changes that occurs in for 5 min to wash the pellet, then the ethanol was the human body is usually caused by the decanted and the DNA air-dried at room substances available in medicinal plants temperature.The pellets was suspended in 100- (Nwachukwu et al., 2010), and the fresh or dried 200 µl of nuclease free water for storage or used.parts of plants can be used to inhibit and cure different types of sicknesses (Joharchi and Amiri, Table 1: Studied Plants Used in the Treatment of 2012).Generally, the utilization of medicinal Malaria plants differs from species to species, as illnesses changes from one pattern to another in different sites (Nwachukwu et al., 2010).Nevertheless, at the market level medicinal plants and their parts are usually contaminated or changed by other plants thereby causing loss of value and hazard due to physical relatedness of plant part that causes misidentification by users and absence of standard identification system (Joharchi and Amiri, 2012).
Random amplified polymorphic DNA marker system is simple, fast, cheap and widely used for genetic diversity studies and marker assisted selection.Considering the poverty and Delhi.The bands were scored on the basis of D N A E x t r a c t i o n u s i n g C TA B M e t h o d presence (1) or absence (0).Fresh young leaf tissue (100 g) was weighed and grinded with 1000 µl of extraction buffer, vortexed and poured into sterile tube of 1.5 ml.It Polymerase Chain Reaction (PCR) Analysis o was incubated in water bath at 60 C for 10 min, PCR amplification was performed in and equal volume of phenol, chloroform, and isovolume of 25 µL which consisted of 20 ng of amylacohol at 25: 24: 1 was added.It was genomic DNA, 2.5 µl of 10 x buffer Inqaba vortexed and centrifuged at 1, 3000 g for 10 min Biotechnical Industries (Pty) Ltd.Pretoria, and 450 µl of the supernatant was removed and South Africa, 1.5 µl of 50 mM MgCl (Inqaba 2 poured into 1.5 ml sterile tube and 400 µl of cold Biotechnical Industries (Pty) Ltd.Pretoria, isopropanol was added into the supernatant.It o South Africa) 2.0 µl of 2.5 mM dNTPs (Inqaba was mixed and incubated for 3 h at 20 C in Biotechnical Industries (Pty) Ltd.Pretoria, freezer and centrifuge at 1,3000 g for 10 min to South Africa), and 0.2 µl of 500U Taq DNA sediment the DNA.The supernatant was decanted to ensure that the pellets was not polymerase (Inqaba Biotechnical Industries distributed and 500 µl of 70% ethanol was added (Pty) Ltd.Pretoria, South Africa), 1.0 µl of 10 µM to the pellets and centrifuged again at 1,3000 g of each primer and 15.05 µl DEPC-treated water for 5 min to wash the pellet, then the ethanol was (invitrogen corporation).The PCR cycling profile decanted and the DNA air-dried at room used was an initial step at 94°C for 5 min., 35 O temperature.The pellets was suspended in 100cycles at 94°C for 20 sec, 60 C for 30 sec, 72°C 200 µl of nuclease free water for storage or used.
for 1 min, and a 10 min final extension at 72°C.Amplified products were resolved on 2% agarose Electrophoresis of the Extracted Genomic DNA gel, visualized under UV light and photographed.The extracted DNA was separated in 1.5% agarose gel. 2 µl of the 6X DNA loading dye Results and Discussion was mixed with 5 ìl of the extracted DNA and the Banding Profiles Obtained using RAPD Markers mixture was loaded into the gel wells.10 µl of The data obtained from the scoring of the 100 base pair DNA ladder was used as molecular amplicons were used for phylogenetic size marker and was loaded into the first well and reconstruction using unweighted pair group ran alongside with the samples at 100 volts for 1 mean arithmetic (UPGMA) and dissimilarity index h in the electrophoresis tank.The gel was viewed in Jaccard's option (Jaccard, 1908) were used for under UV trans-illuminator and the image the analysis.The analysis was conducted using captured using gel documentation system BR NTSYSpc software version 2.02.Biochem.Life Science LTD, Tilak Nagar, New   the generated amplicons gave a representative Phytomed.2: 105 -112. of the accessions.However, this result is in accordance with the report of Yook et al. (2014) Kamatenesi-Mugisha, M., D. W., Makawiti, H., who observed that RAPD studies have been Oryem-Origa, and Olwa-Odey. (2005).widely used for genetic variation of population.
Ethnopharmacological screening of Vernonia The results of the study showed that RAPD amygdalina and Cleome gynandra traditionally marker used established remarkable genetic used in child birth in western Uganda.Pp. 110difference among the medicinal plants 122 in Proceedings of the 11th Natural Product examined.
Research Network for Eastern and Central Africa (NAPRECA) Symposium.Antananarivo,