Department of Biotechnology and Biological Sciences

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http://hdl.handle.net/20.500.11837/156

The Department of Biotechnology and Biological Sciences collection in DSpace at the University of Fort Hare provides access to the scholarly and research outputs of staff and students. This collection includes research articles, conference papers, and examination question papers. The Department integrates a broad range of disciplines, including Biochemistry and Microbiology, Botany, Zoology, Entomology, Biotechnology, Molecular Biology, and Genetics, to provide comprehensive teaching and research in the life sciences. Its programmes and research activities address critical challenges in health, agriculture, biodiversity conservation, food security, industry, and the environment, while promoting both fundamental and applied scientific inquiry. By preserving and disseminating these resources, the collection supports teaching, learning, and research, while contributing to scientific advancement and sustainable solutions for local and global communities.

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The allelopathic potential of Arctotis Arctotoides (L.f.) O. Hoffm on some vegetables
(University of Fort Hare, 2012) Badmus, Abimbola Adesile; Afolayan, A J
Laboratory and greenhouse experiments were conducted to evaluate the allelopathic effects of the extracts and residue Arctotis arctotoides (L.f.) O. Hoffm on selected vegetable crops. The study aimed to address the following specific objectives to (i) examine the ultra structures of the leaf of A. arctotoides using the Scanning Electron Microscope (SEM), (ii) carry out comprehensive qualitative and quantitative phytochemical analysis of the root and shoot materials of the plant, (iii) investigate the allelopathic activities of the root and shoot aqueous extracts of A. arctotoides at concentrations of 10, 8, 6, 4 and 2 mg/ml on germination, radicle and plumule growth of cabbage, carrot, tomato and spinach, (iv) evaluate the inhibitory effects of the dried shoot residue of the plant at 10, 20 and 40 g kg-3 of soil (treatments B, C and D) and the control (treatment A) on the morphology, growth and chlorophyll pigment content of tomato and cabbage transplants at 1, 2, 3 and 4 weeks after transplanting and (v) assess the effects of the dried shoot residue of A. arctotoides on the yield, nutrient uptake by the leaves of tomato and cabbage at 4 and 12 weeks after transplanting. Finally, to analyze the residual mineral content of the soils with tomato and cabbage transplants at 12 weeks after transplanting. The the SEM revealed that anisocytic stomata and glandular trichomes (GTs) were numerous on the abaxial than the adaxial surfaces of A. arctotoides. The non glandular trichomes (NGTs) were also present on both surfaces but lesser on the abaxial. Morphologically, the GTs were peltate, uniseriate and globular head while the NGTs were cylindrical and filamentous with variable number of cells at the basal portion. The energy dispersive X-ray spectroscopy of some crystals showed that Na+ Mg2+ and Ca2+ were the major constituents of the crystal deposit found around the GTs and stomata. The results of the phytochemical composition of the root and shoot extracts of A. arctotoides confirmed the occurrence of alkaloids, flavonoids, phenolics, saponnins, tannins and triterpenes as the common constituents. In addition, cardiac glycosides and steroids were also detected in the shoot of the A. arctotoides. Quantitative estimation of the chemical constituents of the crude extracts further revealed that the alkaloid content in the root higher (0.97 percent) than the shoot (0.64 percent). The quantity of flavonoids detected in the shoot (1.02 percent) was more than that observed in the root (0.35 percent). Others (phenolics and tannins) were marginal in the two plant parts. The results of the inhibitory effects of the root and shoot aqueous extract at the varying concentrations showed that root extract at 10 mg/ml considerably reduced the germination of cabbage, carrot, tomato and spinach seeds by 84.0, 83.2, 72.8 and 37.4 percent respectively. Incubation of the shoot extract at the same concentration resulted in 100 percent inhibition of cabbage and carrot seed germination whereas those of tomato and spinach were suppressed by 91.5 and 61.2 percent respectively. The two extracts at the varying concentrations also had a significant reduction on the radicle and plumule growth of the four vegetables. Addition of the shoot residue to the soil showed massive chlorosis, necrotic lesions and wilting of tomato and cabbage leaves under treatments C and D at 2 weeks after transplanting. The number of leaves, leaf area, dry shoot and root weight of the two vegetables grown in the amended soils were also drastically reduced. The inhibition percentages due to the addition of the three concentrations of A. arctotoides dried shoot residue on the dry shoot weight at 4 weeks after transplanting were 38.6, 45.5 and 70.3. for tomato and 57.5, 73.3 and 87.5 percent for cabbage. Similarly, the declines in the dry root weight of 61.3, 82.9.4 and 83.4 percent for tomato as well as 53.1, 54.7 and 67.2 percent for cabbages were recorded for the two vegetables under treatment B, C and D during the period. The results further showed that the dry fruit yield and shoot weight of tomato under the treatments B, C and D decreased with increase in shoot residue concentrations of A. arctotoides. Relative to treatment A, no significant differences were recorded in the dry head weight of cabbage under the residue treated groups. The reductions in the fruit yield and fresh head weight caused by treatments C and D were 37.2 and 84.8 percent for tomato and 30.9 and 72.4 percent for cabbage. The findings on the mineral contents in the leaves of the two vegetables revealed significant differences in the uptake of N, Mg, Na, Cu and Fe by tomato leaves. The concentrations of N, K, Na and Zn in cabbage leaves also differed. However, the P content was relatively constant in the leaves of the two vegetables at 4 and 12 weeks after transplanting. At 12 weeks after transplanting, the Fe content in soils with tomato and cabbage treatments C and D was greatly enhanced in comparison with the other nutrients. The residual N, P and Zn detected in soils planted to cabbage were similarly equal among all the groups including the control. Thus, under the greenhouse experiment, Arctotis arctotoides (L.f) O. Hoffm has been shown to contain some phytotoxic chemical compounds in its root and shoot materials. The compounds either singly or collectively have demonstrated some inhibitory potentials on the germination, growth and yields of cabbage, carrot, tomato and spinach evaluated in this study.
Antibacterial properties of the methanol extract of helichrysum pedunculatum
(University of Fort Hare, 2008) Ncube, Nqobile S; Okoh, Anthony I; Afolayan, A J
The methanol extract of Helichrisum pedunculatum was screened for antimicrobial activity up to a concentration of 5 mg/ml using the agar dilution technique. A number of test bacterial isolates, comprising both Gram negative and Gram positive organisms were susceptible to the crude extract of the plant. The minimum inhibitory concentrations (MICs) of the extract ranged between 1 and 5 mg/ml for the susceptible organisms. The MICs of the selected antibiotics, chloramphenicol and penicillin, ranged between 2 and 4 mg/L, and 2 and 32 mg/L respectively against Bacillus cereus, Proteus vulgaris and Staphylococcus aureus OKOH1. Bactericidal activity was determined by the time kill assay. The methanol extract of the plant was not bactericidal at 1 × MIC for B. cereus, P. vulgaris and Staph. aureus OKOH1. At 2 × MIC the extract was bacteriostatic against B. cereus but bactericidal against P. vulgaris and Staph. aureus OKOH1. Combination studies were done at 1/2 × MIC, 1 × MIC and 2 × MIC of the plant extract with 1 × MIC of the antibiotics. Combinations of the plant extract and chloramphenicol resulted in mostly indifferent interactions against P. vulgaris and Staph. aureus OKOH1 but synergistic interactions at higher concentration of the plant extract for B. cereus. Penicillin combinations gave synergistic interactions at lower concentrations of the plant for P.vulgaris and Staph. aureus OKOH1 but was mostly indifferent for B. cereus.
Biochemical evaluation of Tulbaghia violacea harv.rhizomes in diet induced hypercholestrolemic rats
(University of Fort Hare, 2012) Olorunnisola, Olubukola Sinbad; Afolayan, A J
Discovery of cheap, nontoxic and readily available antiatherosclerotic drugs is an extraordinary challenge in this modern world. Atherosclerosis and cardiovascular diseases have been predicted to be the leading cause of death by the year 2030. Hence, this thesis was designed to search for plant (s) with anti-atherogenic properties, investigate its possible side effects and extrapolate its likely mechanism(s) of action. An ethnobotanical survey was employed in identification of locally important plants used for the management and treatment of cardiovascular diseases and its predisposing factors in Nkonkobe Municipality, Eastern Cape in South Africa. Information on the names of plants, their parts used and methods of preparation was collected through a questionnaire which was administered to herbalists, traditional healers and rural dwellers. The most frequently used plant (Rhizomes of Tulbaghia violacea Harv.) was investigated for toxicity using brine shrimp lethality (in vitro) and in vivo toxicity test (acute and subchronic) on rats to determine safety dosage. The in vitro antioxidant and free radical scavenging activity of the plant was investigated using models such as 1,1-diphenyl-2- picrylhydrazyl (DPPH), superoxide anions, hydrogen peroxide (H2O2), nitric oxide (NO), 2,2’- azinobis [3-ethylbenzothiazoline-6-sulfonic acid] diammonium salt (ABTS), lipid peroxidation inhibition and the ferric reducing agent. Phytochemical content and the effect of oral administration of fresh methanolic extract rhizomes of Tulbaghia violacea (250, 500 mg/kg. bwt/day) on Lipid peroxidation (TBARS), serum and tissue antioxidant enzymes in normal, hypercholesterolemic and diet induced atherogenic rats were also assessed. More so, the potential of the extract (250 and 500 mg/kg. bwt) to protect against atherogenic diet (4 percentage cholesterol 1 pecentage cholic acid and 0.5 percentage thiouracil) induced fatty streaks formation, dyslipidemia, oxidative stress and endothelial dysfunction was also investigated. Ethnobotanical study revealed that 19 plant species are used for the treatment of heart related diseases in the Municipality. 53 percentage of the plants mentioned were used for the management of chest pain, 47 percentage for high blood pressure, 42 percent for heart disease, 16 percentage for stroke and 11 percentage for the treatment of hypercholesterolemia. Tulbaghia violacea was repeatedly mentioned as the plant species used for the treatment of high blood pressure and predisposing factors in the study area. The brine shrimp cytotoxicity test revealed that fresh, dried methanolic extracts and essential oil of the T. violacea exhibited a high degree of cytotoxic activity with IC50 values of 18.18 (fresh) and 19.24 (dried) μg/ml. An IC50 value of 12. 59 μg/ml was obtained for the essential oil of the plant. The low cytotoxicity values obtained, suggested that rhizome of T. violacea may serve as a potential source of antimicrobial and anticancer agents. In vivo acute study of single oral administration of 5g/kg dose does not produce mortality or significant behavioral changes during 14 days observation. In the sub-chronic study, the extract (250, 500 mg/kg/bwt/ day) administered for a period of 28 days showed no mortality or morbidity. The weekly body and organ weight of the rats showed no significant differences between the control and the rats treated with the extract. The extract at all doses does not show any effect on of biomarkers of liver or renal damage. However, a significant decrease in the activity of ƔGT was observed in the extract treated groups. Hematological evaluation revealed that oral administration of fresh methanolic extracts of rhizomes of T. violacea does not cause anaemia or leucocytosis in the animals. Furthermore, histopathology results of the internal organs revealed no detectable inflammation. These results demonstrated that the rhizome extract of T. violacea was potentially safe for consumption orally even in chronic concentration. In vitro antioxidant evaluation showed that the essential oil, fresh and dried methanolic extracts exhibited potent antioxidant activities in a concentration dependent manner. Phytochemical investigation reveals that the fresh and the dry extract of RTV are rich in flavonoid, flavonol, phenols, tannin and proanthocyanidin, while the essential oil contained dimethy disulfide, dimethyl trisulfide, (methyl methylthio) methyl, 2,4-dithiapentane (11.35 percent) and (methylthio) acetic acid, 2- (methylthio) ethanol, 3-(methylthio) - and propanenitrile (7.20 percent). The fresh extract had higher radicals scavenging activity than the essential oil or dried extract, with 50 percentage inhibition of DPPH, hydrogen peroxide and lipid peroxidation at a concentration of 35.0 ± 0.12, 19.3 ± 0.11 and 17.9 ± 0.15 μg/ml respectively. Oral administration of methanolic extract of RTV in 125, 250 and 500 mg/kg to female Wistar rats significantly inhibited reduction of glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT). The extracts also inhibited (p< 0.05) lipid peroxidation in normal, high cholesterol and diet induced atherosclerosis fed rats in a dose dependant manner. Also the extract (250 and 500 mg/kg/bwt/day) caused a significant (p<0.05) improvement in body weight of treated animals compared with untreated hypercholesterolemia control rats. The extracts also protected significantly (p<0.05) against atherogenic diet induced liver damage or fatty streaks formation in the aorta as revealed by histological examination. The anti-cholesterolemia and anti-atherosclerotic activities of the extract compared favorably well with standard drugs Gemfibrozil and Atorvastatin respectively. Conclusively, rhizomes of T. violacea possess significant anti-atherogenic activity and its mechanism of action(s) may be due to its antioxidant and anti-hypercholesterolemia properties. The results of this study also suggested that rhizome of T. violacea is relatively safe for human consumption and it may be used as an alternative to garlic.
Comparative phytochemical analyses of Aloe Ferox Mill. found in Eastern and Western Cape provinces in South Africa
(University of Fort Hare, 2013) Adams, Zanele; Afolayan, A J
Development of conservation methods for gunnera perpensa l.: an overexploited medicinal plant in the Eastern Cape, South Africa
(University of Fort Hare, 2014) Chigor, Chinyere Blessing; Afolayan, A J
South Africa, many plants which are used in traditional medicines are collected from wild populations. The high demand for trade and use of these medicinal plants place an enormous pressure on their natural populations, especially because they are indiscriminately harvested. The most affected of these plant species are those harvested from their underground parts, among which is Gunnera perpensa L. Gunnera perpensa is of considerable ethnobotanical interest in traditional medicine because of its wide usage. The rhizomes are widely used and indiscriminately collected in large quantities from the wild to meet the ever increasing demand in traditional medicine markets. As a result, this valuable medicinal plant species is being endangered. According to the Red List of South African Plants, the conservation status of G. perpensa has been listed as ‘declining’. The ethnobotanical survey conducted as part of this research confirms the plant species as threatened. It is, therefore, important to develop propagation and conservation strategies for this medicinal plant. Clonal propagation of G. perpensa was conducted using varying lengths of the rhizome (1, 2, 3, 4 and 5 cm) segments as propagules. While regeneration was possible with all the rhizome lengths, most of the growth parameters were significantly higher in the 5 cm rhizomes than the other rhizome segments. The appropriate planting depth for the rhizomes was also determined and 4 or 5 cm planting depths were found appropriate. No significant difference was observed in the growth parameters amongst the planting depths; nevertheless, 4 cm depth gave higher growth and yield. The results of this study show that regenerating G. perpensa.
Heavy metal content absorption and medicinal potential of Egeria densa (Planch.) Casp
(University of Fort Hare, 2013) Mgobozi, Vuyokazi; Afolayan, A J
The contamination of heavy metals in the environment is a looming concern worldwide. Egeria densa (Planch) (Submerged aquatic plant) from two ponds: Site A with co-ordinates (32º 48’22.04”S; 26°48’58.79” E) and Site B with co-ordinates (32°48’33.25”S; 26°48’33.25”S) in Alice (Eastern Cape) was evaluated for its ability to absorb heavy metals, phytochemical constituents, antimicrobial activity and ultra-structure using standard analytic procedures. Cadmium (Cd), copper (Cu), iron (Fe), lead (Pb), manganese (Mn), and zinc (Zn) were measured in water, sediments and plant. The concentrations of these metal elements were determined with use of Inductively Coupled Plasma- Optical Emission Spectrometry (ICP-OES). In sediments, the heavy metals (mg/kg) decreased in the order of their average concentration as follows: Fe (40.320) > Zn (1.259) > Pb (0.564) > Mn (0.186) > Cu (0.037) in Pond 1 whereas in Pond 2 Fe (61.527) > Cd (0.999) > Mn (0.648) > Pb (0.586) > Zn (0.156) > Cu (0.045). The highest concentration of Fe was detected in both sites and Cu being the least. The concentrations of the metals in the plants sample (from Pond 1) were found in order of Mn > Pb > Cu > Fe whereas cadmium and zinc were not detected, while the concentration in Pond 2 decreases in order of Zn > Mn > Pb > Cd > Fe > Cu. In the water samples, concentrations of heavy metals (mg/L) decreased in the order of their average concentrations as follows: Pb (35.36) > Fe (3.07) > Mn (0.238) > Cu (0.104), both cadmium and zinc were below the limit of detection in Pond 1, whereas in Pond 2 the concentrations decreased as follows: Pb (13.033) >Fe (1.69) > Cu (0.270) > Mn (0.248) > Cd (0.004) and Zinc was not detected. Phytochemical analyses of the plant extracts revealed the presence of phenols, flavonoids, proanthocyanidin, flavonols, saponins, alkaloid and tannins in all the extracts (water, acetone and n-hexane). Both acetone and water extracts, showed high concentration of proanthocyanidin, while tannin was the lowest in acetone extract. Antimicrobial evaluation using, Gram positive (Staphylococcus aureus, Bacillus pumilus, Bacillus cereus, Streptococcus pyogenes, Enterococcus faecalis) and Gram negative (Klebsiella pneumonia, Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Serratia marcescens) bacteria showed negative results for all the strain, except Streptococcus pyogenes which was inhibited at MIC of 0.1 mg/ml. Scanning electron microscopy (SEM) of ultra-structure of Egeria densa, showed that certain bacteria attached to the leaf, However more work has to be done on E. densa to verify the mechanism by which it accumulates heavy metals. The study shows that E. densa has a potential of accumulating heavy metals especial Manganese in plant.
Investigation of antidiabetic properties, mechanisms of action and toxicology of Strychnos Henningsii (GILG) bark
(University of Fort Hare, 2011) Oyewole, Oyedemi Sunday; Afolayan, A J; Bradley, G
The apparent reversal of trend from modern drugs to herbal medicine is partly due to the fact that synthetic drugs have always shown adverse reactions and other undesirable side effects. Hence, the use of medicinal plants for the treatment of diseases such as diabetes is very common especially in the rural areas. Majority of these plants are used based on the experience and indigenous knowledge without identification of the therapeutic agents. There is enormous wealth of medicinal plants in the world yet many of them have not been discovered or studied scientifically to substantiate their ethno-medicinal usages. Ethnobotanical study has been the method often used to search for locally important plant species for the discovery of crude drugs with low side effects. An ethnobotanical survey was conducted on the medicinal plants commonly used for the management of diabetes mellitus in Nkonkobe Municipality, Eastern Cape of South Africa. Information was obtained through structured questionnaire administered to traditional healers and herbalists in the region. The study revealed 15 species of plants belonging to 13 families. Strychnos henningsii and Leonotis leonorus among others were repeatedly mentioned by the traditional healers as the two mostly used plants for the management of diabetes mellitus. The infusion and decoction of the roots, leaves and barks of these plants are the methods of preparation. The antioxidant potential of aqueous bark extract of S. henningsii was investigated both in vivo and in vitro using spectroscopic method. The antioxidant activity of the extract against hydrogen peroxide (H2O2), 2,2′-azinobis[3-ethylbenzothiazoline6-sulfonic acid] diammonium salt (ABTS), as well as reducing power was concentration dependent. The extract exhibited lower and average scavenging activities against 1,1diphenyl2picrylhydrazyl (DPPH) and nitric oxide (NO) radicals with IC50 value of 0.739 and 0.49 mg/ml respectively. The administration of the plant extract at 250, 500 and 1000 mg/kg significantly increased the activities of the antioxidant enzymes in the hepatotoxic rats induced with carbon tetrachloride. On the other hand, the stem bark extract had lower effect on lipid peroxidation level except at the dose of 250 mg/kg. The effect of oral administration of S. henningsii extract was evaluated in normal Wistar rats for 28 days. The observed result indicated non- toxic effect of sub-acute administration of plant extract to the animals except at certain doses. This is because, there was no apparent damage to some haematological and biochemical parameters used in assessing organ specific toxicity. However, the alterations observed on platelet, white blood cells and its differentials imply parameter and dose selective toxicity when repeatedly consumed on daily basis at the doses investigated. This study also investigated the antidiabetic activities of the extract at the doses of 125, 250 and 500 mg/kg body weight in diabetic rats induced with streptozotocin -nicotinamide for 15 days. The extract appreciably (P <0.05) reduced the blood glucose level, feed and water intake while the best result was obtained at 250 mg/kg. Similarly, the level of triacylglycerol at the three doses investigated was significantly decreased. In addition, the glucose tolerance was reduced to near normal level after 90 min at certain doses. The clinical significance of the extract on some biochemical and haematological parameters lessen both hepatic and renal damages. Anaemic condition in diabetic animals was also improved after plant extract administration. However, no significant effect was observed in white blood cells and some of its differentials. The extract demonstrated strong glucose utilization in 3T3-L1 cells with a response of 278.63 percent of the control at 12.5μg/ml while that of Chang liver cells was 103.54 percent. The cytotoxicity result revealed non toxic effects of the extract to both cell lines. Treatment of 3T3 L1 cells with the extract did not reduce lipid accumulation. The extract inhibited the activity of α- glucosidase and α- amylase in a concentration dependent manner with IC50 values of 38 μg/ml and 60.9 μg/ml respectively. The percentage protein antiglycation of S. henningsii was 18.4, 38.2 and 61.2 perceent for 0.25, 0.5 and 1 mg/ml respectively while aminoguanidine a known inhibitor of protein glycation was 87.2 percent at 1 mg/ml. The FRAP assay values of the extract was 357.05 μmol Fe (II)/g. The findings from this study support the folkloric usage of this plant for the management of diabetes mellitus in the region.
Phytochemical analysis and antibacterial properties of aqueous and ethanol extracts of Brachylaena elliptica (Thurb.) dc. and Brachylaena ilicifolia (Lam.) Phill & Schweick
(University of Fort Hare, 2015) Sagbo, Idowu Jonas; Afolayan, A J; Bradley, G
Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.
Phytochemical analysis and bioactivity of selected South African medicinal plants on clinical isolates of Helicobacter pylori
(University of Fort Hare, 2011) Njume, Collise; Ndip, R N; Afolayan, A J
Medicinal plants have been used as traditional medicine in the treatment of numerous human diseases for thousands of years in many parts of the world. In the developing world, especially in rural areas, herbal remedies continue to be a primary source of medicine. Scientifically, medicinal plants have proven to be an abundant source of biologically active compounds, many of which have already been formulated into useful therapeutic substances or have provided a basis for the development of new lead molecules for pharmaceuticals. Antibiotic resistance, undesireable side effects and expences associated with the use of combination therapy in the treatment of Helicobacter pylori infections have generated a considerable interest in the study of medicinal plants as potential sources of new drugs against this organism. The high complexicity of bioactive compounds accumulated in plants coupled with their broad antimicrobial activity may make it difficult for pathogenic organisms, including H. pylori to acquire resistance during treatment. This study therefore evaluates the antimicrobial potential of selected South African medicinal plants employed in the treatment of H. pylori-related infections, and the subsequent isolation of the plant active principles. An ethnobotanical survey of plants used in the treatment of H. pylori-related infections was conducted in the study area. Crude extracts of Combretum molle, Sclerocarya birrea, Garcinia kola, Alepidea amatymbica and 2 Strychnos species were screened against 30 clinical strains of H. pylori and 2 standard control strains (NCTC 11638 and ATCC 43526). In the preliminary stages of this study, ethyl acetate, acetone, ethanol, methanol and water extracts of the plants were tested against H. pylori by agar well diffusion and micro broth dilution methods. The plant crude extracts that exhibited anti-H. pylori activity with a iv percentage susceptibility of 50 percent and above were considered for the rate of kill assays and the most active crude extracts selected for bio-assay guided isolation of the active ingredient. Preliminary fractionation of the crude extract was achieved by thin layer chromatography (TLC) using different solvent combinations; hexane/diethylether (HDE), ethyl acetate/methanol/water (EMW) and chloroform/ethyl acetate/formic acid (CEF) in order to determine the most suitable combination for column chromatography (CC) and subsequent testing by indirect bioautography. The extract was then fractionated in a silica gel column using previously determined solvent combinations as eluent. Active fractions obtained from column chromatography separations were further fractionated and the compounds identified by gas chromatography/mass spectrometry (GC/MS) analysis. All the plants exhibited antimicrobial activity against H. pylori with zone of inhibition diameters ranging from 0 - 38 mm and minimum inhibitory concentration (MIC) values ranging from 0.06 - 5.0 mg/mL. The most active plant extracts were the acetone extract of C. molle with a percentage susceptibility of 87.1 percent, acetone and aqueous extracts of S. birrea (71 percent each) and the ethanolic extracts of G. kola (53.3 percent). Except for the aqueous extract, these extracts also exhibited a strong bactericidal activity against H. pylori at different concentrations. TLC analysis revealed the presence of 9 components in the acetone extract of S. birrea with the EMW solvent system as opposed to 5 and 8 with HDE and CEF respectively. Bioassay-guided isolation led to the identification of 52 compounds from the acetone extract of S. birrea with n-octacosane being the most abundant (41.68 percent). This was followed by pyrrolidine (38.91 percent), terpinen-4-ol (38.3 percent), n-eicosane (24.98 percent), cyclopentane (16.76 percent), n-triacontane (16.28 percent), aromadendrene (13.63 percent) and α-gujunene (8.77 percent). Terpinen-4-ol and pyrrolidine demonstrated strong antimicrobial activity against H. pylori at all concentrations tested. These results may serve as preliminary scientific validation of the ethnomedicinal uses of the above mentioned plants in the treatment of H. pylori-related infections in South Africa. Terpinen-4-ol and pyrrolidine could be considered for further evaluation as therapeutic or prophylactic agents in the treatment of H. pylori-related infections. However, further investigations would be necessary to determine their toxicological properties, in-vivo potencies and mechanism of action against H.pylori
Phytochemical screening and thin layer chromatographic profiling of aloe vera (l) burn. f growing in South Africa
(University of Fort Hare, 2013) Dubeni, Zimasa Busisiwe; Afolayan, A J
The chemical profiling, characterization of Aloe products and phytochemical properties of Aloe vera were studied. The adulteration of commercial products derived from medicinal plants has been a major muddle for both the society and the pharmaceuticalindustry. Economically motivated adulteration includes the potential for contaminated, sub‐potent or counterfeit medication to enter the supply chain at several levels, from the production of raw ingredients through to the point of retail sale. Darwin’s theory of evolution states that, species undergo genetic variation with time to adapt to environmental changes. Therefore, the same species growing in widely different habitats may drift from the original genetic makeup as a mechanism of adaptation and that may result in them having different chemical profiles. Therefore this study aimed at investigating the phytochemical properties of Aloe vera growing in South Africa. Also, this study aims to utilize Thin Layer Chromatography to profile this plant, as well as use Infra Red spectroscopy to characterize commercial Aloe vera products. A large quantity of Aloe vera plant was collected from AloeWay, Iphofolo Game Farm, Polokwane in the Limpopo province of South Africa. The identity of the plant was confirmedrom literature and authenticated by Professor DS Grierson of Botany Department, University of Fort Hare, Alice. The plant leaves were divided into two portions. One portion was extracted fresh while the other was cut into pieces and oven dried at 400C then and milled to a homogenous powder once dried completely. The phytochemical composition of the gel and leaf extracts revealed the presence of alkaloids, flavonoids, saponins, tannins and phenols at different concentrations. Results showed that the dry plant material yielded more phytochemicals than the fresh plant material. In particular, it was found that the acetone extract showed much more amounts ofphychemicals than the dichloromethane and aqueous extracts. The percentage compositions of phenols (71.86), flavonols (36.61), proanthocyanidins (82.71), saponins (37.73) and alkaloids (13.29) were significantly high in the acetone extract, followed by the dichloromthane extract with values of 46.85, 37.73, 49.51, 89.0 and 11.11 respectively, while the least composition was found in the aqueous extract. Furthermore, flavonoids were somewhat high in composition in both the aqueous extract of the dried and of the fresh plant material while others were very low. Tannins levels were significantly very low in all the solvent extracts. It was found that the acetone extract showed great amounts of phytochemicals than dichloromethane and aqueous extracts. Since A. vera is used in the treatment of different ailments such as skin wounds and abrasions, eczema, constipation, rheumatoid arthritis etc, the medicinal uses of this plant could be associated to such analysed bioactive compounds. Acetone, hexane, ethanol, water and dichloromethane were used to extract the Aloe vera leaf and the best solvent extract was determined. Thin layer chromatography was used to profile the leaf extracts with the aim of documenting the main phytochemicals present in the Aloe vera growing in South Africa. The best spraying reagent was determined. Fourier transform infrared spectrophotometer was used to validate the presence of Aloe vera ingredients in commercial products. The yield extraction ability of the solvent was the order: water>ethanol> hexane >dichloromethane and acetone for the dry portion. However, for the plant extracted fresh, the order of yield produced was ethanol-acetone-dichloromethane > and water. The different solvent systems separated the compounds differently. Hexane: acetone: ethanol (20 : 5: 2) and Benzene: ethanol: ammonium (80): ethanol (10): ammonium solvent systems were noted to be the best mobile phase as they gave the best separation compared to other systems.EMW [ethyl acetate (81): methanol (11): water (8)] showed better separation than the other two separating solvent systems. Vanillin- sulphuric acid spray was seen to be the best spraying reagent as compared to vanillin- phosphoric acid. Fourier transform infrared spectrophotometer validated the presence aloe ingredients in aloe vera commercial products.
Studies on the use of essential oils for the control of Sitophilus Zeamais (Motschulsky) (Coleoptera; Curculionidae): a pest of stored maize grains
(University of Fort Hare, 2008) Odeyemi, Oluwakemi Oluwaseyi; Afolayan, A J
The common maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), a well known pest of stored-maize grain in most parts of the world, was identified as one of the major constraints of harvested maize grains in the Eastern Cape Province, South Africa. The use of plants or their products is one of the recent methods being investigated for insect pest control worldwide. Therefore, the main aim of the present study was to identify available plants in the Eastern Cape that could be used to combat the problem of Sitophilus zeamais in stored-maize grains. For the realization of the aims of this research, the following studies were carried out; a preliminary survey was conducted to obtain baseline information on the farmers’ knowledge and experience of indigenous insect pest control methods in the Eastern Cape. Also, studies on the insecticidal potential of the essential oils of some plants were investigated against the maize weevil. The quality parameters of maize grains treated with the essential oils was also studied and, using a rat model, the toxicity of the essential oils was investigated. The outcome from this study revealed that there is awareness amongst the farmers in the Eastern Cape on the use of plants or their products to control insect pests. Unfortunately, such methods are currently being neglected and the knowledge of their application was found to be eroding. Among the various essential oils screened were those from Mentha longifolia L. and Tagetes minuta L. which evoked an appreciable level of contact, fumigant and repellent toxicity on the maize weevil. Further work done to determine the effects of the oils on maize stored over a period of three months revealed that the two oils had no adverse effect on the proximate compositions and some quality parameters of the stored maize. However, the toxicological study conducted on rats showed that the oils at tested concentrations exhibited some level of toxicity. It is, therefore, suggested that the essential oils of M. longifolia and T. minuta should not be used to treat maize grains intended for human consumption.
Validating the use of Hippobromus pauciflorus (L.f) Radlk for the treatment of eye infections in the Eastern Cape South Africa
(University of Fort Hare, 2010) Pendota, Srinivasa Chary; Afolayan, A J; Grierson, D S
An ethnobotanical survey of plants used for the treatment of eye infections was carried out in the Eastern Cape Province of South Africa. The study revealed that 12 plant species are commonly used with Hippobromus pauciflorus (L.f) Radlk being the most frequent. The crude extracts of the leaves, stem bark and roots of the plant were investigated for antimicrobial activity against 10 bacterial and four fungal strains. The methanol extracts of the plant parts were the most active and showed appreciable activity against Gram-positive and Gram-negative bacteria. The methanol extracts of the leaves and stem bark inhibited the growth of fungi with activities ranging from 78.70 percent to 100 percent on Aspergillus niger and Penicillium notatum. The acetone extracts of the leaves and stem bark were active against A. niger (51.76 percent) and P. notatum (77.22 percent). The aqueous extract of Hippobromus pauciflorus leaves at 50, 100 and 200 mg/kg body weight doses were evaluated for anti-inflammatory, analgesic and antipyretic activities in male Wistar rats. Anti-inflammatory activity was studied by using carrageenan-and histamine induced oedema right hind paw volume while the analgesic effect was evaluated using formalin-induced pain and tail flick nociception response. The brewer’s yeast-induced pyrexia model was used for the antipyretic investigation. The extract at all the doses used significantly inhibited both the carrageenan- and histamine-induced inflammation in a manner that was not dose dependent. The extract reduced the formalin-induced pain licking as well as prolonged the reaction time in the tail flick-induced pain. The effects of the aqueous extract of the plant leaves at 50, 100 and 200 mg/kg body weight doses were investigated for 14 days on some biochemical parameters of male Wistar rats. The extract at all the doses tested did not significantly alter the levels of white blood cells, red blood cells, mean corpuscular volume, platelets, neutrophils, monocytes, lymphocytes and large unstained cells. While the levels of haemoglobin, packed cell volume and basophils increased at specific doses, those of mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration and eosinophils decreased. In vitro antiviral activities of the plant against herpes simplex virus type 1(HSV-1) and coxsakie virus B6 were investigated. Cytotoxicity was evaluated by MTT assay in Vero cells. At concentrations ranging from 165-270/μg/ml there was significant antiviral activity against HSV-1, but toxicity was also noted. There was no antiviral activity against coxsakie virus B6. Bioactivity-guided fractionation of the leaves of H. pauciflorus yielded three known compounds. From the ethyl acetate fraction, two compounds were isolated (epicatechin and -sitosterol ) and from the n- hexane fractions, one compound (lupeol) was isolated. They were isolated and identified using various techniques. The antimicrobial, anti-inflammatory, analgesic and anti-pyretic activity of these compounds have been reported in literature. The structure and distribution of foliar appendages on the leaves of H. pauciflorus were examined by scanning electron microscope. The leaves have long unicellular nonglandular trichomes which were distributed over the mid rib and densely populated at the edges of the adaxial and abaxial surfaces. In general, the experiments and tests conducted in this study appear to have justified the use of Hippobromus pauciflorus for the treatment of eye infections and make a substantial contribution to the knowledge base of the use of herbal medicine for the treatment of the eye infections.

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