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This study was designed to assess the cytotoxicity, anti-HIV and antibacterial efficacy of various solvent extracts of roots, stem and leaves of Alchornea laxiflora, as well as five compounds isolated from its methanolic stem extract viz.; ellagic acid (1); 3-O-methyl-ellagic acid (2), 3-O-β- d -glucopyranosyl-β-sitosterol (3), 3-O-acetyl-oleanolic acid (4) and 3-O-acetyl-ursolic acid (5). The tested crude extracts were prepared from several solvent polarities including: hexane (Hex), chloroform (CHCl3), ethyl acetate (EtOAc), ethanol (EtOH), methanol (MeOH) and water (H2O). The anti-HIV properties were assessed on HIV-1 subtype C integrase while the cytotoxicity was tested against Hela cells. The antibacterial activity was studied on a panel of pathogens including gastrointestinal, skin, respiratory and urinary-tract infection causing Gram positive bacteria viz.; Bacillus cereus (ATCC 11778), Enterococcus faecalis (ATCC 29212), Staphylococcus aureus (ATCC 25923) and Staphylococcus saprophyticus (ATCC 15305)] and Gram-negative bacteria, i.e., Escherichia coli (ATCC 25922), Klebsiella pneumoniae (ATCC 13883), Moraxella catarrhalis (ATCC 23246). All the tested samples were determined to be non-toxic due to the low inhibitions observed. The most potent anti-HIV activity was observed for the methanolic extract of A. laxiflora root (ALR4) with an IC50 value of 0.21 ng/ml, which was more active than chicoric acid used as reference drug (6.82 nM). Roots, stem and leaves of A. laxiflora extracts exhibited antibacterial activities against most of the Gram-positive bacteria with the minimum inhibitory concentrations (MIC) ranging between 50 and 63 μg/ml. Compounds 1–5 displayed antibacterial activities against S. saprophyticus with MIC values as low as 4 μg/ml. The results inferred from this study demonstrate the potential of A. laxiflora root as a source for new anti-HIV drugs and scientifically validate the traditional use of A. laxiflora in the treatment of gastrointestinal, skin, respiratory and urinary tract related infections. These results reaffirm the ethnopharmacological significance of African traditional medicines.

Rapanea melanophloeos is traditionally used in South Africa in the treatment of ailments of the skin, pulmonary and gastro intestinal tract. This study was aimed at giving an overview of these traditional uses and comparing the phytochemicals and antibacterial activities of various crude extracts of the leaves, fruits and bark in order to validate these uses. The three plant parts were extracted using petroleum ether (PE), ethyl acetate (EtOAc), methanol (MeOH) and water. Various phytochemicals were compared using TLC, while alcohol precipitable solids (APS), non-polar terpenes and amino acids were analysed by GC-MS. Antibacterial activity was determined against three Gram-positive and three Gram-negative strains by microdilution assays. Caryophyllene oxides, α-cadinol and (−)-spathulenol were identified in the PE extracts. All nine essential amino acids were present in fruit extracts in significantly higher levels than in the leaves and bark; 255.1, 23.4 and 21.3 mg/g respectively. Most of the extracts showed good antibacterial activity, especially against the Gram-positive pathogens (MIC of ≤1 mg/mL), the EtOAc extracts exhibited the best activity with the fruit having an MIC values of 0.1 ± 0.2 mg/mL against Staphylococcus epidermidis and Enterococcus faecalis, 0.05 mg/mL against Bacillus cereus. Results from this study validate the ethnomedicinal uses of R. melanophloeos extracts for ailments of bacterial etiology. The plant had a rich supply of secondary metabolites, APS and amino acids and TLC and antibacterial activities of the extracts showed slight variations in chemical composition due to geographic distribution.

The one-pot synthesis of silver nanoparticles (AgNPs) using the medium-polar extract of Desmodium adscendens (Sw.) DC. is presented here as an alternative synthesis of metal NPs. Characterisation of the formed NPs showed polydispersed AgNPs ranging from 15 to 100 nm where the concentration of metal ions was found to play a role in the size and shape of the prepared NPs. It could be established that the flavonoids, saponins, and alkaloids present in the extract acted as both reducing and stabilising agents during the formation of the capped metal NPs. This means of NP synthesis was also employed during the in situ immobilisation of AgNPs on gauze and plaster. An evaluation of the antibacterial activity of the medium-polar D. adscendens extract, AgNPs suspended in solution, and the immobilised AgNPs against Staphylococcus aureus (ATCC 25923), Bacillus cereus (ATCC 11778), and Escherichia coli (ATCC 25922) showed high efficacy against the latter in particular. This suggests that gauze, dilute silver nitrate solutions, and D. adscendens extract could be used successfully in the simple in situ preparation of effective antibacterial wound dressings.

Ethnopharmacological relevance Tetracera potatoria Afzel. Exg. Don (Dilleniaceae) is a medicinal plant used traditionally in Africa for the treatment of tuberculosis related ailments and respiratory infections. The antibacterial activity of the medium polar extracts of T. potatoria leaves and stem bark was recently reported against Mycobacterium smegmatis (MIC 25 µg/mL) and M. aurum (65 µg/mL), two fast-growing Mycobacterium strains used as model micro-organisms for the more pathogenic strain Mycobacterium tuberculosis ( Fomogne-Fodjo et al., 2014 ). The aim of this study was consequently to isolate the compounds possibly contributing to this activity, and which may therefore be promising precursors to be used for the development of novel anti-TB drugs. Materials and methods T. potatoria medium polar extract [MeOH/DCM (1:1, v/v)] was fractionated sequentially with petroleum ether to which EtOAC and MeOH were gradually added to increase the polarity. The examination of T. potatoria extract and its fractions was guided by bioassays for anti-mycobacterial activity against M. smegmatis (ATCC 23246) and M. aurum (NCTC 10437) using the minimum inhibitory concentration (MIC) method. All the isolated compounds were structurally elucidated using spectroscopic techniques and evaluated for their anti-mycobacterial activity. Results Two novel secondary metabolites (1, 2) named tetraceranoate and N-hydroxy imidate-tetracerane, together with five known compounds [β-stigmasterol (3), stigmast-5-en-3β-yl acetate (4), betulinic acid (5), betulin (6) and lupeol (7)] were isolated and identified. Tetraceranoate exhibited the best activity against M. smegmatis with a minimum inhibitory concentration (MIC) of 7.8 µg/mL, while β-stigmasterol, betulinic acid and betulin showed appreciable anti-mycobacterial activity against both strains (MIC 15 µg/mL). Conclusion Seven compounds were isolated from the medium polar extract [MeOH/DCM (1:1, v/v)] of T. potatoria stem bark. Only tetraceranoate one of the isolated compounds showed antibacterial activity against M. smegmatis having efficacy as high as rifampicin (one of a three drug regimen recommended in the initial phase short-course anti-tuberculosis therapy). Thus, tetraceranoate might be an interesting target for systematic testing of anti-TB treatment and management. This research supports the use of T. potatoria in African traditional medicine for the treatment of tuberculosis related symptoms.

Ethnopharmacological relevance The leaves, stems and roots of Alchornea cordifolia (Schumach. and Thonn.) Mull. Arg. are used as traditional medicine in many African countries for the management of gastrointestinal, respiratory and urinary tract infections as well as for the treatment of wounds. Aim of the study To determine the in vitro antibacterial activity of the crude extracts of leaves and stems of A. cordifolia on gastrointestinal, skin, respiratory and urinary tract pathogens and to identify the compounds in the extracts that may be responsible for this activity. Materials and methods The antibacterial activities of crude extracts [hexane, chloroform (CHCl 3 ), ethyl acetate (EtOAc), ethanol (EtOH), methanol (MeOH) and water (H 2 O)] as well as pure compounds isolated from these extracts were evaluated by means of the micro-dilution assay against four Gram-positive bacteria, i.e. Bacillus cereus ATCC 11778, Enterococcus faecalis ATCC 29212 , Staphylococcus aureus ATCC 25923 and S. saprophyticus ATCC 15305 , as well as four Gram-negative bacterial strains, i.e. Escherichia coli ATCC 25922 , Klebsiella pneumoniae ATCC 13883 , Moraxella catarrhalis ATCC 23246 and Proteus mirabilis ATCC 43071. The isolation of the active constituents was undertaken by bio-autographic assays in conjunction with chromatographic techniques. The identification and characterisation of the isolated compounds were done using mass spectrometry (MS) and Fourier transformed infrared spectrometry (FTIR) as well as 1D- and 2D- nuclear magnetic resonance (NMR) analyses. Results The leaves and stems of A. cordifolia exhibited varied antibacterial activity against all eight pathogens. Most of the MIC values ranged between 63 and 2000 µg/ml. The highest activities for the crude extracts (63 µg/ml) were observed against S. saprophyticus [stem (EtOAc, CHCl 3 and hexane), leaves (MeOH, EtOH, EtOAc and CHCl 3 )], E. coli [stem (MeOH and EtOH), leaves (MeOH, EtOH, EtOAc and CHCl 3 )], M. catarrhalis [leaves (EtOAc and CHCl 3 )], K. pneumoniae [stem (CHCl 3 ), leaves (CHCl 3 )] and S. aureus [leaves (CHCl 3 )]. Seven constituents [stigmasterol ( 1 ), stigmasta-4,22-dien-3-one ( 2 ), friedelin ( 3 ), friedelane-3-one-28-al ( 4 ), 3- O -acetyl-aleuritolic acid ( 5 ), 3- O -acetyl-erythrodiol ( 6 ) and methyl-3,4,5-trihydroxybenzoate (methyl gallate) ( 7 )] were isolated from the stem MeOH extract. All these compounds displayed some antibacterial activity against the eight pathogens with highest activity against S. saprophyticus (2 µg/ml). Furthermore, this is the first report of compounds 1 , 2 , 3 , 4 , 6 and 7 isolated from A. cordifolia and where a complete set of 2D-NMR data for fridelane-3-one-28-al ( 4 ) is presented. Conclusion The study demonstrated that the antibacterial activities of A. cordifolia extracts may be due to the presence of the seven isolated compounds, where compounds 3–6 showed the best activity. The observed activity against gastrointestinal, skin, respiratory and urinary tract pathogens supports the traditional use for the treatment of such ailments.

Ethnopharmacological relevance Annickia affinis (Exell) Versteegh & Sosef, closely related to A. chlorantha Setten & P.J.Maas (both species also referred to as Enantia chlorantha Oliv.), from the Annonaceae family, are multi-purpose medicinal plants used widely across tropical Africa. The two Annickia species are morphologically distinct from each other and have different distribution patterns, but are frequently confused. Furthermore, the name Enantia chlorantha is an illegitimate name, but is still used today. Materials and methods A review of the literature was undertaken and an in-depth analysis of previous research and future prospectives are considered. While a myriad of publications cite the species “ Enantia chlorantha ”, this is not the case for A. affinis and A. chlorantha, and no reviews are available for any of the species to date. Consequently, a summary of their ethnobotany, phytochemistry and biological properties is presented here (for the period 1933 – November 2014) in order to substantiate their traditional importance as medicines for rural people in Africa. Results To this effect, these species seem to be the preferred traditional treatments for malaria in tropical Africa, an area suffering heavily under the malaria pandemic. Their chemical composition is dominated particularly by various isoquinoline alkaloids, as well as by acetogenins and sesquiterpenes, which have been isolated from the bark and leaves. All three of these classes of compounds have been reported to exhibit noteworthy biological activity. Conclusions Due to their widespread use, especially of the bark, these species have already been categorized as threatened with extinction. Consequently this study further aims to identify areas where more research needs to be conducted involving these important species, and also to suggest possible means of increasing the biological activities of their extracts as a way to conserve the species.

The 100 K structures of two salts, namely 2-amino-1H-benzimidazolium 3-phenylpropynoate, C7H8N3+·C9H5O2−, (I), and 2-amino-1H-benzimidazolium oct-2-ynoate, C7H8N3+·C8H11O2−, (II), both have monoclinic symmetry (space groupP21/c) and display N—H...O hydrogen bonding. Both structures show packing with corrugated sheets of hydrogen-bonded molecules lying parallel to the [001] direction. Two hydrogen-bonded ring motifs can be identified and described with graph setsR22(8) andR44(16), respectively, in both (I) and (II). Computational chemistry calculations performed on both compounds show that the hydrogen-bonded ion pairs are more energetically favourable in the crystal structure than their hydrogen–bonded neutral molecule counterparts.

Elephantorrhiza elephantina (Burch.) Skeels (Fabaceae) and Pentanisia prunelloides (Klotzsch ex Eckl. & Zeyh.) Walp. (Rubiaceae) are two medicinal plants used extensively in southern Africa to treat various ailments. Often, decoctions and infusions from these two plants are used in combination specifically for stomach ailments. The antimicrobial activities of the methanol and aqueous extracts of the rhizomes of the two plants, as well as the two active ingredients from the plants [(−)-epicatechin and palmitic acid] have been determined apart and in combination against Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 8739) and Bacillus cereus (ATCC 11778). The minimum inhibitory concentration (MIC) values for the aqueous (0.50–16.00 mg/mL) and methanol (0.20–16.00 mg/mL) extracts independently demonstrated varied efficacies depending on the pathogen of study. When the two plants were combined in 1:1 ratios, synergistic to additive interactions (ΣFIC values 0.19–1.00) were noted. Efficacy for the two major compounds ranged between 0.13–0.63 mg/mL and mainly synergistic interactions were noted against E. faecalis and E. coli . The predominantly synergistic interactions noted between E. elephantina and P. prunelloides and major compounds, when tested in various ratios against these pathogens, provide some validation as to the traditional use of these two plants to treat bacterial gastrointestinal infections.

The main diterpenoids in the roots of Arctopus (Apiaceae, tribe Saniculeae) have been identified for the first time. The major compounds are manool, ent- trachyloban-19-oic acid and a kauren-19-oic acid, while methyl-16β-hydroxy- ent -kaur-11-en-19-oate is a minor diterpene. Comparative studies of non-polar extracts by means of LC–MS showed that the isolated main compounds are present in all three species of Arctopus. The study also revealed that the diterpenoid pattern in Arctopus is very similar to that of the genus Alepidea . The tuberous roots of Arctopus and the rhizomes and roots of Alepidea are important traditional medicines in South Africa, used mainly for infections and respiratory ailments. Their efficacy is ascribed to antimicrobial and anti-inflammatory activity. The known biological activities of the isolated diterpenes provide a scientific rationale for the traditional uses. The chemical similarity in diterpenoids also supports the idea that the two genera are closely related, despite their conspicuous morphological differences.

The occurrence of (R)-3′-O-β- d -glucopyranosylrosmarinic acid, rosmarinic acid and caffeic acid in two important South African medicinal plants is reported for the first time. (R)-3′-O-β- d -Glucopyranosylrosmarinic acid and rosmarinic acid were isolated and identified in several samples from three species of the genus Arctopus L. (sieketroos) and three species of the genus Alepidea F. Delaroche (ikhathazo), both recently shown to be members of the subfamily Saniculoideae of the family Apiaceae. The compounds occur in high concentrations (up to 15.3 mg of (R)-3′-O-β- d -glucopyranosylrosmarinic acid per g dry wt) in roots of Arctopus. Our results provide a rationale for the traditional uses of these plants, as the identified compounds are all known for their antioxidant activity, with rosmarinic acid further contributing to a wide range of biological activities. Furthermore, we confirm the idea that (R)-3′-O-β- d -glucopyranosylrosmarinic acid is a useful chemotaxonomic marker for the subfamily Saniculoideae.

The structure of a new cycloartane-type triterpene glycoside was determined as 24,25- O- β - d -diglucopyranosyl-6α-hydroxycycloart-3-one (SU3) by spectroscopic methods. This is the first cycloartane diglycoside reported from the genus Sutherlandia (an important South African traditional medicine and general tonic known as cancer bush). It was isolated from a dwarf form of S. frutescens , currently known as Sutherlandia humilis .

Ethnopharmacological relevance The antibacterial activities of 18 plants from 10 different families were investigated for their antimicrobial efficacy, based on the traditional uses of these species by Bakola pygmies living in Central Africa, especially along the Ngoyang area in Cameroon for the treatment of respiratory and tuberculosis-related symptoms. The aim of the study is to test the antimicrobial efficacy of these plants against some pathogens associated with respiratory disease and to determine if there is any validation for the traditional use against Mycobacterium species. Materials and methods Medium polar extracts were prepared in MeOH/DCM (1:1, v/v) from the plant parts of each species used traditionally and were assayed against pathogens associated with respiratory tract ailments [ Staphylococcus aureus (ATCC 25923), Klebsiella pneumoniae (ATCC 13883) and Morexella cattarhalis (ATCC 14468)] using the minimum inhibitory concentration (MIC) method. Two additional faster growing Mycobacterium strains [ Mycobacterium smegmatis (ATCC 23246) and Mycobacterium aurum (NCTC 10437)] were included in the assay as predictive test organisms for the more pathogenic strain Mycobacterium tuberculosis. Results Some plant species, such as Alchornea floribunda , Musanga cecropioides (both leaves and stem bark), Tetracera potatoria and Xylopia aethiopica (stem bark), were effective in inhibiting Morexella cattarhalis , having MIC values between 65 and 250 μg/mL. Some noteworthy antimycobacterial inhibition (MIC≤200 μg/mL and as low as MIC 6.5 µg/mL) for 54% of the extracts were observed. Conclusion While moderate activity was shown for pathogens causing respiratory tract infections, these plant species seems to be selectively targeting Mycobacteria spp. suggesting that the traditional use for treating tuberculosis related symptoms may be indeed be accurate.

Ethnopharmacological relevance Alchornea floribunda Mull. Arg. is used in traditional medicine across Africa for the treatment of bacterial, fungal, parasitic and inflammatory disorders. Aim of the study To evaluate the antibacterial activity of the crude extracts of different plant parts in order to provide a scientific rationale for the proposed broad efficacy of Alchornea floribunda in the treatment of bacterial infections. Materials and methods Extracts of roots, stems and leaves were prepared using solvents of various polarities in order to extract a wide range of phytochemicals. The antibacterial activity of these crude extracts was evaluated by micro-dilution assay, against Gram-positive (i.e. Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus and Staphylococcus saprophyticus) as well as Gram-negative (i.e. Escherichia coli, Klebsiella pneumoniae, Moraxella catarrhalis and Proteus mirabilis) bacteria. Results Generally, the ethanol (EtOH), methanol (MeOH), ethyl acetate (EtOAc) and chloroform (CHCl3) extracts demonstrated the best activities, with the leaves exhibiting the highest average activity for six of the eight pathogens. Of these, the ethanolic leaf extract was the most active against Staphylococcus aureus with an MIC value of 50 µg/mL. Some other notable activity was observed for the ethyl acetate and chloroform root extracts against Staphylococcus aureus (50 µg/mL), and for selected stem extracts against Staphylococcus aureus (50 µg/mL), Klebsiella pneumoniae (63 µg/mL) and Staphylococcus saprophyticus (63 µg/mL). Conclusion This study demonstrates the promising antibacterial activity of Alchornea floribunda against both Gram-positive and Gram-negative bacteria responsible for gastrointestinal, skin, respiratory and urinary ailments, and validates its use in the ethnopharmacology of the region.

Ethnopharmacological relevance Bitterness values have been determined for southern African plant species that are traditionally used as tonics ( imbizas or ’musa-pelo ) to alleviate the symptoms of stress and a variety of ailments related to the digestive system. Aim of the study To measure and present, for the first time, the bitterness values of 15 of the best-known and most widely used tonic plants in southern Africa in order to find a rationale for their traditional use in improving appetite and treating digestive ailments. Results Most of the plants were found to be very bitter, with bitterness values comparable to those reported for internationally well-known bitter tonics such as Artemisia absynthium L. and Gentiana lutea L. Conclusions The relatively high bitterness values obtained for all of the plants indicate that their alleged value in improving digestion and appetite may at least be partly ascribed to the bitter tonic ( amarum ) effect, i.e., the stimulation of gastric juices via the nervus vagus . It may be interesting to examine the chemical compounds responsible for the bitter taste, as well as the possible links between bitterness and the anecdotal anti-stress properties ascribed to these species.

Lippia javanica (N.L.Burm.) Spreng. is an aromatic, multipurpose medicinal plant from which a number of volatile compounds have been identified, together with toxic triterpenoids and iridoid glycosides. Two additional phenylethanoid glycosides, verbascoside and isoverbascoside, were isolated from L. javanica and characterized. High performance liquid chromatography analyses of polar extracts of three other Lippia species (L. scaberrima, L. rehmannii and L. wilmsii), indigenous to South Africa, revealed the presence of both isomers. When compared to the other indigenous Lippia species, the leaves of L. javanica were found to contain the highest concentrations of both isomers. In addition, the intraspecies variation of the verbascoside/isoverbascoside content of L. javanica, harvested from the same and different localities, was investigated. The concentrations of the two phenylethanoids remained fairly consistent within and between different populations, even when geographically separated. While these compounds are produced by many genera, they may now be added to the list of iridoid glucosides employed as chemotaxonomic markers for Lippia species.