Open Access Original Research Article

Development of Sequence Characterized Amplified Region (SCAR) Marker for the Authentication of Bacopa monnieri (L.) Wettst

Aradhana Yadav, Javed Ahmad, Anis A. Chaudhary, Altaf Ahmad

European Journal of Medicinal Plants, Page 186-198
DOI: 10.9734/EJMP/2012/1192

Aims: To develop Sequence Characterized Amplified Region (SCAR) marker for identification of Bacopa monnieri (L.) Wettst.
Study design: Molecular biology tools for authentic identification of Bacopa monnieri.
Methodology: RAPD-based SCAR marker was developed to identify Bacopa monnieri from its adulterant candidates namely Centella asiatica, Eclipta alba and Malva rotundifolia. 50 random primers were used for initial screening of different accessions of Bacopa monnieri, Eclipta alba and Malva rotundifolia. A putative 589 bp marker specific to Bacopa monnieri was identified using RAPD technique. This RAPD-amplicon was then sequenced and cloned. Based on the information of cloned sequences a pair of SCAR primers was designed. SCAR primers were then used for authentication of DNA samples of Bacopa monnieri and its adulterants. Market samples of Bacopa monnieri and Centella asiatica collected under the name of Brahmi was put to test with these primers.
Results: Out of 50 random primers, only 14 primers were able to amplify the above plants. A 589 bp polymorphic band obtained with OPAA-3 primer which was specific to Bacopa monnieri accessions and not found in other adulterant candidates was selected. This band was eluted, cloned and further sequenced. A pair of SCAR primers (Bac F & Bac R) between 406 bp of 589 bp sequence of RAPD amplicon was designed. A single, bright, distinct band was obtained in Bacopa monnieri and not in the adulterants. Further validation was also done in the market samples.
Conclusion: In essence, the study was to develop a RAPD-based SCAR marker for authentication of Bacopa monnieri. The SCAR marker was found to be useful for preventing the adulteration of other plants in Brahmi and also for screening of crude drug samples intended for export and domestic uses.

Open Access Original Research Article

Changes in Erythrocytic Membrane Free Energy of Albino Rabbits Administered Ethanol Leaf Extract of Spondias mombin Linn

C. U. Igwe, C. G. Osuagwu, V. A. Onwuliri, G. O. C. Onyeze

European Journal of Medicinal Plants, Page 199-208
DOI: 10.9734/EJMP/2012/1062

Aims: To determine the effect of ethanol extract of Spondias mombin Linn on membrane potential energy, the intracellular and extracellular concentrations of Na+, K+, Ca2+ and Mg2+ in an easily accessible model cell, erythrocyte, was studied. The effect on the first approximation estimates of erythrocytic free energy changes (ΔG) was calculated.
Study Design: Randomized study.
Place and Duration of Study: Department of Biochemistry, Federal University of Technology, Owerri, Nigeria between January 2010 and February 2011).
Methodology: Twenty-five female albino rabbits (1.47 ± 0.17 kg) of the same age set were divided into 5 groups of 5 animals. The first group served as the baseline control, the groups II and III animals were intraperitoneally administered 1ml of 750 mg/kg body weight of S. mombin extract daily for 7 and 14 days respectively, while groups IV and V animals were intraperitoneally administered 1ml of 0.14 IU/kg body weight of oxytocin drug ((Pitocin®, USA) daily for 7 and 14 days respectively. Plasma and lyzed red blood cells obtained were analyzed for extracellular and intracellular erythrocyte concentrations of sodium, potassium, calcium and magnesium using standard methods.
Results: The study showed that both the extract and oxytocin administrations significantly (p<0.05) reduced intracellular [Mg2+] and extracellular [K+] and [Ca2+], with a concomitant increase in intracellular [Ca2+]. Both treatment with the extract and oxytocin, did not significantly (p>0.05) alter [Na+] and extracellular [Mg2+]. On the other hand, while the extract did not affect intracellular [K+], oxytocin significantly (p<0.05) reduced it. The results also indicated that while the extract significantly (p<0.05) increased the [K+]-based estimated ΔG, oxytocin treatment reduced it. On the other hand, both the extract and oxytocin caused a non-significant (p>0.05) drop in the Ca2+ based ΔG, with no significant (p>0.05) alteration of the [Na+] and [Mg2+] based estimated ΔG. The pole reversal observed with the study of the Ca2+-based ΔG indicates depressive effect of both the extract and oxytocin on Ca2+ ATPase pump.
Conclusion: The observations above indicate that, while the use of the extract might enhance energy generation and conservation, the use of oxytocin might elicit exhaustive utilization of energy.

Open Access Original Research Article

Potential Antimicrobial Plant Extract Based Therapeutics from Temnocalyx obovatus Roots

P. Dzomba, N. Muchanyereyi

European Journal of Medicinal Plants, Page 209-215
DOI: 10.9734/EJMP/2012/1293

Aims: The study was conducted to determine antimicrobial activity of Temnocalyx obovatus mature root extracts popularly used in folk medicine to treat diarrhoea in chickens, stomach disorders in turkeys, goats and cows, snakebites, asthma, ulcers and whooping cough in humans.
Study Design: Agar disk diffusion method and determining MIC.
Place and Duration of Study: Department of Chemistry (Natural product section) June 2011 and July 2011.
Methodology: Two methods were employed for the determination of antimicrobial activities; an agar paper disc diffusion method and determination of minimal inhibitory concentration (MIC). Methanol and ethanol extracts were assayed for antimicrobial activities. The following bacterial strains were employed in the screening studies: Staphylococcus aureus, Escherichia coli, Clostridium perfringens and fungi: Aspergillus niger, Candida albicans all from stock cultures of Bindura University department.
Results: Methanol extract exhibited the greatest extraction efficiency; 136,1g/Kg as compared to ethanol; 124,8 g/Kg. Both methanol and ethanol extracts showed significant antibacterial and antifungal activity. Sensitivity to the extracts was not similar for the chosen strains. The highest antibacterial activity of plant extract was 37,0 mm for methanol and 28,0 for ethanol extracts for diameter of zone inhibition found against Clostridium perfringens followed by 35,0 mm and 26,3 diameter of zone inhibition against Escherichia coli at a concentration of 100%. Antifungal activity was highest for Aspergillus niger at a concentration of 100%. The minimum inhibitory concentration (MIC) values against these bacteria ranged from 10 - 60 µg/ml. In comparison to reference standards Gentamicin, and miconazole, methanolic extracts showed significant antimicrobial activity, student t-test, p = 0,05.
Conclusion: The present results support the use of aqueous extracts of Temnocalyx obovatus roots in folk medicine for the treatment of conditions such as diarrhoea and stomach disorders. These results encourage follow-ups through bioassay-directed isolation of the phytocompounds responsible for the potency.

Open Access Original Research Article

Comparison of Carbohydrate Structures and Immunomodulating Properties of Extracts from Berries and Flowers of Sambucus nigra L.

Hilde Barsett, Torun H. Aslaksen, Parakastha Gildhyal, Terje E. Michaelsen, Berit Smestad Paulsen

European Journal of Medicinal Plants, Page 216-229
DOI: 10.9734/EJMP/2012/1335

Aims: To investigate if the immunomodulating activity of compounds present in berries and flowers of S. nigra were of the same order, or different, and also if the most active components were of high or low molecular weight nature.
Methodology: Defatted material of berries and flowers of S. nigra were extracted with 50% ethanol and with water of 50ºC and 100ºC. High molecular weight fractions were obtained after gel filtration on BioGelP6DG. The different fractions were investigated for their monosaccharide contents and carbohydrate structures. The immunomodulating effects were investigated using a complement fixing assay as well as a system for measuring the production of NO after stimulation of macrophages with the different fractions.
Results: All fractions contained substantial amounts of carbohydrates. Removal of low molecular weight material revealed polysaccharide fractions containing monosaccharides typical for pectins and showed enhanced bioactivity.
Conclusion: High molecular weight fractions from elderflowers showed higher bioactivity than the equally extracted fractions from elderberries. The 100ºC water flower fraction gave after gel filtration the fraction with the highest activity and with the longest backbone of rhamnogalacturonan I. Based on this; one should encourage the drinking of juice made form flowers in the same way as has been done for the juice made from berries, since both may have a beneficial heath effect.

Open Access Original Research Article

Pharmacological Activities of a Mongolian Medicinal Plant, Malva mohileviensis Down.

G. Odontuya, G. Enkhmaa, N. Batbayar, R. Naran, K. T. Inngjerdingen, T. E. Michaelsen, B. S. Paulsen

European Journal of Medicinal Plants, Page 230-241
DOI: 10.9734/EJMP/2012/1280

Aims: To investigate the Mongolian plant M. mohileviensis Down. for the presence of possible bioactive products that could be related to the traditional use of the plant in Mongolia.
Methodology: Ethanolic and water extracts of both seeds and herb were tested for anti-inflammatory and DPPH scavenging activity. Polysaccharides were isolated from the seeds using sequence of chromatographic methods. The polysaccharide fraction from the cold water extract was then analyzed for the presence of monosaccharides and their type of linkages by GC and GC-MS. The effect of the polysaccharides on the complement system was then determined.
Results: The ethanolic and aqueous extracts of seeds and herbs, as well as crude polysaccharides from cold and hot water extracts exhibited a significant anti-inflammatory activity in the model based on histamine-induced paw edema. Anti-inflammatory effects of all samples were high during the 3rd hour of inflammation. Moreover, the ethanolic extracts of seeds and herbs exhibited DPPH scavenging activity. Phytochemical studies of the cold water seed polysaccharide revealed the presence of an uncommon pectic type polysaccharide. Galacturonic acid (38%) and rhamnose (30%) were present as the main monosaccharides, and linkage analyses revealed that galacturonic acid was present as terminal, 1→4 and 1→3,4 linked units and rhamnose basically as 1→3 linked units. The complement fixation activity was appr. 15 µg/ml, substantially lower than that of the standard used. The results also indicate strongly that the M. mohileviensis and M. verticillata are two different Malva species.

Open Access Original Research Article

In vitro Antibacterial Activity and Phytochemical Screening of Crude Extracts from Jatropha curcas Linn.

K. Nyembo, N. Kikakedimau, H. Mutambel, N. Mbaya, T. Ekalakala, O. Bulubulu

European Journal of Medicinal Plants, Page 242-251
DOI: 10.9734/EJMP/2012/1233

Aims: This study aims at investigating the antibacterial activity of crude methanolic and aqueous extracts of leaves and root barks of Jatropha curcas against Gram-positive and Gram-negative bacteria isolated from urinary tract infections (UTIs) and to confirm the effective use of this plant against the uropathogenic strains in traditional medicine in Democratic Republic of Congo (D.R.C).
Study Design: Laboratory experimental tests; Extraction of J. curcas leaf and root bark, susceptibility tests (zones of inhibition) and minimum inhibitory concentration (MIC) determination and phytochemical screening and high performance liquid chromatography (HPLC) analysis.
Place and Duration of the Study: Department of Phytobiology, Department of Biotechnology and Department of Microbiology, General Atomic Energy Commission. Regional Center of Nuclear Studies of Kinshasa P.O BOX 868 Kin. XI DRC during October and November 2011.
Methodology: Fresh leaves and root barks of J. curcas were collected, oven- dried at 45ºC, powdered and extracted with water and methanol. The aqueous extracts were lyophilized. Agar disc diffusion method was used to test antibacterial activity of the crude extracts of J. curcas against Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Citrobacter diversus isolated from UTIs. The extracts were subjected to phytochemical tests. HPLC method was used to screen phenolic compounds.
Results: The crude extracts exhibited a significant antibacterial activity against four of seven tested bacterial isolates. MIC values ranged from 1.0 to 7.5 mg/L. The extracts phytochemical screening revealed the presence of saponins, tannins, alkaloids, steroids and flavonoids. The presence of phenolic compounds was screened by HPLC analysis.
Conclusion: The inhibitory effects of the crude extracts from leaves and root barks against uropathogenic strains have justified the usefulness of J. curcas for the treatment of UTIs and sexually transmitted Infections (STIs) in traditional medicine of D.R.C.

Open Access Original Research Article

Toxicity of Aqueous Extract of Senna alexandrina Miller Pods on Newzealand Rabbits

Ibrahim B. Elkhidir, Ahmed I. Yagi, Samia M.A. El Badawi, Sakina M. Yagi

European Journal of Medicinal Plants, Page 252-261
DOI: 10.9734/EJMP/2012/1434

Aims: The herbal extract of Senna alexandrina pods is used commonly to treat constipation. The study was designed to evaluate the effect of oral administration of aqueous extract of S. alexandrina pods at varying doses on Newzealand rabbits for 2 and 4 weeks.
Study Design: Biochemical and cross-sectional study.
Place and Duration of Study: Botany Department, Faculty of Science and Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine Science, University of Khartoum.
Methodology: Rabbits of either sex were divided into four groups of 8 animals each. Group I served as control and groups II, III and IV were orally treated with doses of 50, 100 and 300 mg/kg body weight for up to 4 weeks. Toxicity was evaluated using biochemical, hematological and histopathological assays.
Results: No significant pathological changes were observed in rabbits given a dose of 50 mg/kg/day. However, at higher doses alterations in the levels of blood haematological parameters, transaminases, creatinine, albumin and globulin were observed; such changes are likely to occur due to spleen, hepatic and renal injury, which was confirmed by histopathological analysis.
Conclusion: The data suggest that administration of the aqueous extract of S. alexandrina pods at 50 mg/kg/day is not toxic. The observed toxic effect might be due to higher doses and/or frequency of administration. Although in traditional medicine the extract is administrated at a low dose, the results suggest the necessity of standardization of the drug.

Open Access Original Research Article

Ethnomedicinal Study and Screening of Plants Used for Memory Enhancement and Antiaging in Sagamu, Nigeria

T.O. Elufioye, A.T. Oladele, C.M. Cyril-Olutayo, J.M. Agbedahunsi, S.A. Adesanya

European Journal of Medicinal Plants, Page 262-275
DOI: 10.9734/EJMP/2012/1372

Background: The objective of this study was to carry out a comprehensive ethnomedicinal survey of plants used as memory enhancer and antiaging in Sagamu Local Government area of Ogun State in south west Nigeria.
Methodology: Semi-structured questionnaires were used to conduct interviews with traditional healers, herb sellers and other knowledgeable individuals on use of medicinal plants as memory enhancers and anti-aging. Nine communities were selected randomly for data collection across the rural and urban centers with a total of 110 informants.
Results: The informants consist of 92% males and 8% females, 60% were traditional healers, 13% herb sellers while the others were knowledgeable individuals on medicinal plants utilization. Findings revealed 41 plant genera belonging to 31 families of which 46.3%, 26.8%, 17.1% and 12.2% are trees, herbs, climbers and shrubs respectively. Leaves (58.5%) and stem bark (26.8%) are prevalent in plant parts used. Screening of commonly used plants revealed presence of alkaloids, tannins, saponins, anthraquinones, cyanogenetic and cardiac glycosides.
Conclusion: Potentials of plants commonly used by indigenous people in the study area as memory enhancer and antiaging are recommended for further pharmacological research and conservation.