Bhekumthetho Ncube

In vitro antimicrobial synergism within plant extract combinations from three South African medicinal bulbs

ETHNOPHARMACOLOGICAL RELEVANCE Tulbaghia violacea, Hypoxis hemerocallidea and Merwilla plumbea are used in South African traditional medicine for the treatment of some infectious diseases and other ailments. AIM OF THE STUDY The study aimed at investigating the antimicrobial efficacies of independent and various within-plant extract combinations of three medicinal bulbs to understand the possible pharmacological interactions. MATERIALS AND METHODS Bulb and leaf extracts of the three medicinal plants, independently and in combinations, were comparatively assessed for antimicrobial activity against two Gram-positive and two Gram-negative bacteria and Candida albicans using the microdilution method. The fractional inhibitory concentration indices (FIC) for two extract combinations were determined. RESULTS At least one extract combination in each plant sample demonstrated good antimicrobial activity against all the test organisms. The efficacies of the various extract combinations in each plant sample varied, with the strongest synergistic effect exhibited by the proportional extract yield combination of PE and DCM extracts in Merwilla plumbea bulb sample against Staphylococcus aureus (FIC index of 0.1). Most extract combinations demonstrated either a synergistic, additive or indifferent interaction effect against the test bacteria with only a few exhibiting antagonistic effects. CONCLUSION The observed antimicrobial efficacy and synergistic interactions indicate the beneficial aspects of combination chemotherapy of medicinal plant extracts in the treatment of infectious diseases.

A comparative study of the antimicrobial and phytochemical properties between outdoor grown and micropropagated Tulbaghia violacea Harv. plants

AIM OF THE STUDY The study aimed to compare the antimicrobial and phytochemical properties of in vitro cultured and outdoor grown Tulbaghia violacea plants in the quest to validate the use of micropropagated plants as alternatives to outdoor grown plants in traditional medicine. Tulbaghia violacea is used extensively in South African traditional medicine for HIV/AIDS patients and in the treatment of gastrointestinal ailments, asthma, fever and tuberculosis. MATERIALS AND METHODS Extracts of micropropagated and outdoor grown Tulbaghia violacea plants were evaluated for their antibacterial and antifungal activities against Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and a fungus Candida albicans using microdilution methods. Saponins and phenolic compounds including condensed tannins, gallotannins and flavonoids were quantitatively determined using spectrophotometric methods. A qualitative test for saponins was also carried out. RESULTS The petroleum ether (PE) extracts of micropropagated plants and dichloromethane (DCM) extracts of outdoor grown plants showed good antibacterial activity, each against two bacterial test strains. PE extracts of micropropagated plants showed the best antibacterial activity with a minimum inhibitory concentration (MIC) of 0.39 mg/ml against Bacillus subtilis. Good MIC (<1mg/ml) and minimum fungicidal concentration (MFC) values of 0.78 mg/ml were only obtained in DCM extracts of outdoor grown plants. MIC and MFC values for water and ethanol extracts of both micropropagated and outdoor grown plants were similar and in the range 3.125-12.5mg/ml. Total phenolics, gallotannins, flavonoids and saponins were significantly higher in micropropagated plants than in outdoor grown ones. In all cases, the amounts of phytochemical compounds in micropropagated plants were more than twice that of outdoor grown plants except for condensed tannins. CONCLUSION The results form a good basis for the use of Tulbaghia violacea micropropagated plants as a complement to the outdoor grown plants in traditional medicine.

Antioxidant, Antimicrobial and Phytochemical Variations in Thirteen Moringa oleifera Lam. Cultivars

A study was undertaken to assess variation in antioxidant, antimicrobial and phytochemical properties of thirteen Moringa oleifera cultivars obtained from different locations across the globe. Standard antioxidant methods including the DPPH scavenging, ferric reducing power (FRAP) and β-carotene-linoleic acid model were used to evaluate the activity. Variation in the antioxidant activity was observed, with TOT4951 from Thailand being the most active, with activity five times higher than that of ascorbic acid (reference compound). A different trend was observed for the activity in the FRAP and β-carotene-linoleic acid assays. Antimicrobial activity was tested against Gram-positive (Staphylococcus aureus) and Gram-negative (Klebsiella pneumoniae) strains using the microdilution method. Acetone extracts of all cultivars exhibited good antibacterial activity against K. pneumoniae (MIC values of 0.78 mg/mL). The remaining extracts exhibited weak activity against the two microorganisms. For the antifungal activity, all the extracts exhibited low activity. Variations were observed in the total phenolic and flavonoid contents. Cultivars TOT5169 (Thailand) and SH (South Africa) exhibited highest amounts of total phenolic compounds while TOT5028 (Thailand) exhibited the lowest amounts of five times lower than the highest. The information offer an understanding on variations between cultivars from different geographical locations and is important in the search for antioxidant supplementation and anti-ageing products.

Anti-Oxidative and Cholinesterase Inhibitory Effects of Leaf Extracts and Their Isolated Compounds from Two Closely Related Croton Species

A comparative evaluation of the antioxidant and acetylcholinesterase inhibitory activity of the leaf extracts of Croton gratissimus and Croton zambesicus (subgratissimus) and compounds isolated from the extracts was carried out to determine their potential and suitability or otherwise as a substitute for each other in the management of oxidative and neurodegenerative conditions. Different antioxidant assays (DPPH, FRAP, β-carotene-linoleic and the lipid peroxidation models) and the microplate assay for acetylcholinesterase (AChE) inhibition were carried out separately to study the activities of the crude leaf extracts and four solvent fractions from each of the two Croton species. Bioassay guided fractionation was used to target antioxidant constituents of the crude extracts and ethyl acetate fractions of 20% aqueous methanol extract of C. gratissimus on silica gel and Sephadex LH-20 columns resulted in the isolation of kaempferol-3-O-β-6’’(p-coumaroyl) glucopyranoside (tiliroside, 2), apigenin-6-C-glucoside (isovitexin, 3) and kampferol (4). The extract of C. zambesicus yielded quercetin-3-O-β-6’’(p-coumaroyl) glucopyranoside-3’-methyl ether (helichrysoside-3’-methyl ether, 1), kaempferol-3-O-β-6’’(p-coumaroyl) glucopyranoside (tiliroside, 2) and apigenin-6-C-glucoside (isovitexin, 3). Three of the isolated compounds and their different combinations were also included in the bioassays. In all the assays performed, the antioxidant capacity and AChE inhibitory effects of C. zambesicus extracts were weaker than those of C. gratissimus. This suggests that C. gratissimus may not be substituted by C. zambesicus, despite the similarity in some of their constituents. Generally, the combinations made from the isolated compounds showed better activities in most of the assays compared to the individual isolated compounds. This suggests mechanisms such as synergism and/or additive effects to be taking place. This study established low, moderate and high antioxidant activities as well as AChE inhibitory effects by the crude extracts, fractions, compounds and compound combinations. This means some of the extracts, isolated compounds and compound combinations could be useful in the management of neurodegenerative conditions and serve as sources of natural neurodegenerative agents.

Tilting Plant Metabolism for Improved Metabolite Biosynthesis and Enhanced Human Benefit

The immense chemical diversity of plant-derived secondary metabolites coupled with their vast array of biological functions has seen this group of compounds attract considerable research interest across a range of research disciplines. Medicinal and aromatic plants, in particular, have been exploited for this biogenic pool of phytochemicals for products such as pharmaceuticals, fragrances, dyes, and insecticides, among others. With consumers showing increasing interests in these products, innovative biotechnological techniques are being developed and employed to alter plant secondary metabolism in efforts to improve on the quality and quantity of specific metabolites of interest. This review provides an overview of the biosynthesis for phytochemical compounds with medicinal and other related properties and their associated biological activities. It also provides an insight into how their biosynthesis/biosynthetic pathways have been modified/altered to enhance production.

Antibacterial, Antifungal, and Antiviral Activities of African Medicinal Plants

The challenge of disease and death caused by infectious diseases is strikingly apparent in both developed and developing nations around the world, with developing countries being the most affected. As infectious diseases evolve and pathogens develop resistance to existing pharmaceuticals, the search for new novel leads, possibly with different modes of action, against fungal, bacterial, and viral diseases has intensified in recent years. To most of the African population, the use of medicinal plants to combat these challenges forms an integral component of their culture and hence a prominent characteristic. To this extent, several species have been subjected to different pharmacological screening techniques using different pathological strains. As a result, varied ranges of bioactivities have been reported for a number of species, different plant parts, and even geographical location and season. We present here the best reported antimicrobial activities of “Africa’s top 50 medicinal plants.”

Hypoxis (Hypoxidaceae) in African traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE Hypoxis species (Hypoxidaceae) are popular medicinal plants used in the African traditional medicine to treat numerous ailments and are reported to have a wide spectrum of pharmacological properties. In this paper we reviewed and evaluated the traditional uses, pharmacological, phytochemistry and toxicity aspects of the genus Hypoxis. Potential medicinal prospects and possible knowledge gaps and pitfalls are discussed. MATERIALS AND METHODS A comprehensive and systematic review of literature on Hypoxis species was done using numerous resources such as books and scientific databases that include Pubmed, Scopus, Scirus, Google scholar, Web of Science and others. Information on the various pharmacological and chemical properties of Hypoxis extracts was critically analysed and discussed under the various topics. RESULTS The literature indicated a broad range of uses, pharmacological and toxicological properties of different Hypoxis species extracts and their relevance to African healthcare systems. Several compounds, mostly glucosides, sterols and sterolins, have been isolated, identified and tested in various in vitro and in vivo models as well as in a couple of clinical trials with a lot of promising prospects reported in some studies. CONCLUSION A critical analysis of the available literature and studies identifies positive potential for the future use of Hypoxis species in both traditional and modern medicine and concurrently so with possible pitfalls and research gaps in current knowledge. An integrated and holistic approach to addressing research issues, particularly toxicology aspects could be more effectively applied and incorporate conservation strategies for the species.

Antimicrobial, Anthelmintic Activities and Characterisation of Functional Phenolic Acids of Achyranthes aspera Linn.: A Medicinal Plant Used for the Treatment of Wounds and Ringworm in East Africa

Achyranthes aspera Linn. (Amaranthaceae) commonly known as Prickly Chaff flower (English) is traditionally used for treating a number of ailments. Different parts of the plant are used in treating wounds and ringworm in East Africa and elsewhere for a number of ailments. In this study, leaf extracts of A. aspera collected from two different geographical locations (Ciaat, Eritrea and Ukulinga, South Africa) were evaluated for antibacterial, antifungal, anthelmintic activities and the plant characterized for functional phenolic acids as well as protein binding capacity. The pathogens used in the tests were, two Gram-negative (Escherichia coli and Klebsiella pneumoniae), two Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), a filamentus yeast-like fungus (Candida albicans) and a free-living nematode (Caenorhabditis elegans). The water and acetone extracts of the samples collected from Ciaat exhibited good antibacterial, antifungal and anthelmintic activity (MIC < 1 mg/ml) except the water extract against E. coli which showed moderate activity. In contrast, the extracts collected from Ukulinga exhibited moderate to weak activities except for the acetone (aq.) extracts which had good activity against some of the tested organisms. UHPLC-MS/MS revealed variation in the levels of some functional phenolic compounds, with rutin, chlorogenic acid and genistein not being detected in the extracts from Ukulinga. The variation was also observed in the protein binding capacity, which could offer a predictive wound healing model. All extracts from plant samples collected at Ciaat expressed significant dominant potency compared to similar extracts from Ukulinga.

Carbon–nitrogen ratio and in vitro assimilate partitioning patterns in Cyrtanthus guthrieae L

In response to variations in nutritional composition of the growth medium, plants often adjust their metabolism and progressively alter their growth patterns. Carbon (C) and nitrogen (N) constitute the major plant nutritional components influencing plant growth and development patterns. This study examined the growth dynamics and patterns of assimilate partitioning to primary and secondary metabolites in response to varying levels and combinations of C and N in the culture media of Cyrtanthus guthrieae. In vitro callus-derived C. guthrieae plantlets were cultured on solid Murashige and Skoog (MS) media with different concentrations and combinations of C and N. Relative growth rate (RGR) increased proportionally with an increase in C concentrations up to 88 mM sucrose (0.58 d(-1)) beyond which it was hardly influenced by further increases in C. Growth was also significantly favoured in media with high concentrations of N at all C concentrations tested. In C-limited media regimes with growth saturating N conditions, alkaloid accumulation became favoured while polyphenol content increased with an increase in C levels in the medium, a characteristic pattern that appeared to be less influenced by the amount of N. Of the primary metabolites, only proteins showed small significant variations across different media treatments, with starch and soluble sugars increasing proportionately with C levels. In the medium with a high sucrose concentration (175 mM), soluble sugars, amino acids and polyphenols increased markedly, possibly as an adaptive response to the reduced osmotic potential in the media and/or a storage mechanism for excess C and N reserves in the media. From a medicinal perspective, with regard to polyphenolic compounds in C. guthrieae, growth medium conditions that allow for high levels of C pools in the tissue would thus be favourable for the enhanced synthesis of this group of compounds. The medium conditions with 175 mM sucrose and 10.3 mM NH4NO3 gave the highest total polyphenols, flavonoids and proanthocyanidins with a moderate growth rate.

Antimicrobial Activity, Phenolic Content, and Cytotoxicity of Medicinal Plant Extracts Used for Treating Dermatological Diseases and Wound Healing in KwaZulu-Natal, South Africa

Medicinal plants used for wound healing and skin diseases are the key to unlocking the doors to combating problematic skin diseases as resistance of pathogens to pharmaceuticals and allopathic management continues to increase. The study aimed at investigating the antimicrobial efficacies, phenolic content, and cytotoxicity effects of 11 medicinal plant extracts commonly used for treating skin conditions and wound healing in traditional medicine within KwaZulu-Natal. Eleven plant species were separated into different plant parts (bulbs, roots, leaves) and extracted with different solvents. The extracts were assessed for antimicrobial activity against six Gram-positive and seven Gram-negative bacterial strains and four fungi commonly associated with skin conditions using disc diffusion and microdilution techniques. The aqueous methanolic extracts were screened for phenolic content while cytotoxicity tests were performed on all extracts using the brine shrimp lethality and tetrazolium–based colorimetric (MTT) assays. Extracts from Aloe ferox, A. arborescens, and Hypericum aethiopicum were the most active against almost all of the tested bacterial and fungal strains. All plant species exhibited some degree of antimicrobial activity. Total phenolic levels, flavonoids and tannins were also higher for A. ferox, followed by A. arborescens and H. aethiopicum, respectively. The cytotoxicity results of all plant extracts were in the range of 90–100% survival after 24 h in the brine shrimp assay. Extracts considered lethal would demonstrate >50% shrimp death. The MTT cytotoxicity test yielded LC50 values of >1 mg/mL on all extracts indicating that they are not cytotoxic. The observed antimicrobial efficacy demonstrated by some plant species and the general lack of cytotoxic effects on all the tested extracts presents some promising and beneficial aspects of these medicinal plant extracts in the treatment of skin diseases and wound healing. The two Aloe species and H. aethiopicum were among the best extracts that exhibited consistently good antimicrobial activity and warrants further investigations and possible isolation of bioactive principles.