In vitro evidence of phyto-synergy for plant part combinations of Croton gratissimus (Euphorbiaceae) used in African traditional healing

Aim of the study

Despite the extensive traditional use of Croton gratissimus Burch. var. gratissimus for medicinal purposes, scientific studies validating the therapeutic properties of this indigenous plant are lacking. As the bark, roots and leaves of C. gratissimus are used separately as well as in combination, this study focused on determining antimicrobial efficacies of the plant parts independently and in combination to assess possible pharmacological interactions (e.g. synergy, antagonism).

Material and Methods

The hydro-distilled leaf essential oil and extracts of bark, root and leaf were comparatively assessed for antimicrobial activity by means of microdilution minimum inhibitory concentration (MIC). The fractional inhibitory concentrations (FIC) were determined for the leaf and root (1:1), bark and root (1:1), leaf and bark (1:1) combination. Isobolograms were plotted to demonstrate interactions between various ratios of the roots and leaves.

Results

The MIC and FIC results indicated variable efficacies for the various plant part combinations, the greatest of which was noted for Cryptococcus neoformans in the root and leaf combination (MIC 0.4 mg/ml and FIC of 0.4). Isobolograms indicated the greatest synergy for Bacillus cereus, Candida albicans and Cryptococcus neoformans.

Conclusion

The observed synergistic interactions clearly indicate that the reductionist approach may often be short-sighted and that biological activity may be improved through combination therapy, where different complex metabolic pools collectively contribute to the enhanced effect.     

Investigations of antimicrobial activity of some Cameroonian medicinal plant extracts against bacteria and yeast with gastrointestinal relevance

Ethnopharmacological relevance

Resistance against antibiotics of many bacteria is accumulating. Therefore, searches for new substances with antimicrobial activity have become an urgent necessity. Medicinal plants are frequently used in popular medicine as remedies for many infectious diseases (intestinal infection, malaria, tuberculosis, etc.).

Aim of the study

This study aimed to determine the in vitro antimicrobial activity of hexane, ethyl acetate and methanol extracts from different parts (leaves, stem bark, entire plant) of five different plant species against bacteria and yeast of gastrointestinal relevance.

Material and methods

Twenty-one extracts from all the following plants (Albizia gummifera (leaf), Ficus exasperata (leaf and stem bark), Nauclea latifolia (leaf and stem bark), Ricinodendron heudelotii (stem bark), Senna hirsuta (entire plant) have been screened for their antimicrobial activity against eight bacteria species including Gram-positive bacteria (Staphylococcus aureus and Bacillus cereus), Gram-negative bacteria (Escherichia coli, Shigella dysenteriae, Shigella flexneri, Salmonella typhi, Pseudomonas aeruginosa, and Klebsiella pneumoniae) and one yeast species (Candida albicans) using agar disc-diffusion, and microbroth dilution assays.

Results

Results demonstrated that F. exasperata and N. latifolia were active against the whole set of tested microorganisms. The methanol extract of N. latifolia (leaf and stem bark) was the most active against against C. albicans, E. coli, S. aureus, P. aeruginosa with a minimum inhibitory concentration (MIC) of 2, 32, 64 and 64 μg/ml, respectively. The methanol extract of leaf of F. exasperata was also most active with significant inhibitory activity against E. coli, S. dysenteriae, S. Typhi and P. aeruginosa with MIC values of 128 μg/ml. Only the extract of N. latifolia (stem bark and leaf) showed anticandidal property.

Conclusion

The results show that these plant extracts exhibit antimicrobial activity and N. latifolia proved to be most effective as an antibacterial and antifungal.     

Screening of the topical anti-inflammatory activity of the bark of Acacia cornigera Willdenow, Byrsonima crassifolia Kunth, Sweetia panamensis Yakovlev and the leaves of Sphagneticola trilobata Hitchcock

Ethnopharmacological relevance

An investigation of topical anti-inflammatory activity was undertaken on plants used in Central America traditional medicine.

Aim of study

Four herbal drugs used in the folk medicine of Central America to treat inflammatory skin affections (Acacia cornigera bark, Byrsonima crassifolia bark, Sphagneticola trilobata leaves and Sweetia panamensis bark) were evaluated for their topical anti-inflammatory activity.

Materials and methods

Petroleum ether, chloroform and methanol extracts were obtained for herbal medicines and then extracts were tested on Croton oil-induced ear dermatitis model in mice.

Results

Almost all the extracts reduced the Croton oil-induced ear dermatitis in mice and the chloroform ones showed the highest activity, with ID₅₀ (dose giving 50% oedema inhibition) values ranging from 112 μg/cm² (Byrsonima crassifolia) to 183 μg/cm² (Sphagneticola trilobata). As reference, ID₅₀ of the non-steroidal anti-inflammatory drug indomethacin was 93 μg/cm².

Conclusions

Lipophilic extracts from these species can be regarded as potential sources of anti-inflammatory principles.     

Antimicrobial activity of the extracts of Craniotome furcata (Lamiaceae)

Ethnopharmacological relevance

Craniotome furcata (Link.) O. Kuntze (Lamiaceae) is used in the traditional medicine for the treatment of cuts and wounds indicating antimicrobial activity.

Aim of the study

To validate the traditional medicinal claim, in vitro antimicrobial activity of the extracts was screened against eleven human pathogenic bacteria and fungi.

Materials and methods

The ethyl acetate (CE) and n-butanol (CB) extracts of the aerial parts of Craniotome furcata were tested against four Gram-positive bacteria, four Gram-negative bacteria and three fungi species. Antimicrobial activity was determined by the tube-dilution method.

Results

The CE and CB extracts showed antimicrobial activity against all of the tested microorganisms, with minimum inhibitory concentration (MIC) values in the range of 0.41–4.58 mg/ml.

Conclusions

The tested microbes Micrococcus flavus and Escherichia coli were highly susceptible to extract CE and antimicrobial activity of CB is comparable to that of extract CE against Escherichia coli and Staphylococcus faecalis while it is better than that of extract CE, against Klebsiella pneumoniae. Both the extracts were found less susceptible against tested fungi.     

Inhibitory properties of selected South African medicinal plants against Mycobacterium tuberculosis

Ethnopharmacological relevance

Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB) is the most commonly notified disease and the fifth largest cause of mortality. One in 10 cases is resistant to treatment in some areas. Several plants are used locally to treat TB-related disease.

Aims of the study

The aim was to screen selected South African medicinal plants used to treat TB and related symptoms by traditional healers for antimycobacterial activity.

Materials and methods

Ethnobotanical information on these plants was obtained. Crude acetone, methanol, hexane and ethanol extracts of 21 selected medicinal plants obtained in Venda, South Africa were screened for their ability to inhibit MTB H₃₇Ra and a clinical strain resistant to first-line drugs and one second-line drug using tetrazolium microplate assay to determine the minimum inhibitory concentration (MIC). Results were analyzed using Microsoft Excel 2007 and One way ANOVA; p < 0.05 was considered for statistical significance.

Results

Few acetone extracts were active against MTB with MIC under 100 μg/mL. Four plants showed lower MIC values; Berchemia discolor Klotzsch Hemsl 12, 5 μg/mL on H₃₇Ra and 10.5 μg/mL on the clinical isolate, Bridelia micrantha Hochst. Baill (25 μg/mL), Warbugia salutaris Bertol. F Chiov (25 μg/mL), and Terminalia sericea Burch ex D. F (25 μg/mL) on both H₃₇Ra and clinical isolate. However, the roots of Ximenia caffra Sond. Var. caffra, barks of Sclerocarya birrea (A Rich) Hochst, Asclepias fruticosa L, tubers of Allium sativum L, leaves of Carica papaya L, Solanum panduriforme E. Mey C, and roots of Securidaca longepedunculata Fresen gave MIC greater than 100 μg/mL.

Conclusion

The acetone extracts of Berchemiadiscolor, Bridelia micrantha, Terminalia sericea and Warbugia salutaris could be important sources of mycobactericidal compounds against multidrug-resistant MTB.     

Antimicrobial,anti-inflammatory and mutagenic investigation of the South African tree aloe (Aloe barberae)

Ethnopharmacology relevance

In recent times, many products ranging from aloe drinks to aloe gels, powders, capsules, and creams have appeared on the commercial market prepared from different aloe species including Aloe barberae. These products are used in ethnomedicine to treat various conditions including gastrointestinal disorders, insect bites, skin burns and other skin injuries by traditional communities.

Aim of the study

This study was aimed at evaluating the antibacterial, antifungal and anti-inflammatory activities as well as genotoxic effects of different extracts of Aloe barberae.

Materials and methods

Organic and water extracts of the upper stem, young bark, mature bark, leaves and roots of the South African tree aloe (Aloe barberae) were evaluated for their antimicrobial [Gram-positive (Bacillus subtilis and Staphylococcus aureus), Gram-negative (Escherichia coli and Klebsiella pneumoniae) bacteria as well as the fungus Candida albicans], anti-inflammatory (COX-1 and COX-2) and mutagenic properties (Ames test). Thin layer chromatography (TLC) was used to compare the phytochemical profiles of different extracts of Aloe barberae.

Results

The petroleum ether (PE) and dichloromethane (DCM) extracts of the mature bark, leaves and roots exhibited good activity against all the bacteria and fungus Candida albicans with minimum inhibitory concentrations (MIC) ranging from 0.195 to 1.56 mg/ml. All the PE extracts evaluated showed a high activity (>70%) in both COX-1 and COX-2 assays. Apart from the organic extracts of the root with consistently good activity (>70%), all the remaining extracts showed moderate activity (40–69%) in COX-1 assay. The PE extracts also showed a dose dependent increase in activity. Ultraviolet (UV) spectrum of the leaves and root EtOH extracts indicated the presence of compounds that could absorb UV light (wavelength: 190–820 nm). None of the extracts had a mutagenic effect in the Salmonella/microsome assay against a tester strain, TA98.

Conclusion

Activity observed in the bark, leaves and roots of Aloe barberae validates its use in commercial herbal products, ethnobotany and ethnoveterinary medicine by South African communities and small scale farmers to treat various conditions.     

Chemical composition and antimicrobial activity of Phyllanthus muellerianus (Kuntze) Excel essential oil

Ethnopharmacological relevance

Phyllanthus muellerianus (Kuntze) Excel (family Euphorbiaceae) stem bark is used in Cameroon by Baka pygmies as a remedy for wound healing and tetanus.

Aim of the study

To characterize the chemical composition and to evaluate the antimicrobial properties of the essential oil of the plant.

Materials and methods

The essential oil was extracted from the stem bark by dynamic head space and by hydrodistillation and characterized by GC and GC-MS analyses. The antimicrobial activity was evaluated on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal–fungicidal concentration (MBC–MFC) by the micro and macrodilution methods. The following bacteria and fungi were used: Clostridium sporogenes ATCC 3584, Staphylococcus aureus ATCC 6538, Streptococcus mutans ATCC 25175, Streptococcus pyogenes ATCC 19615, Escherichia coli ATCC 10536, Candida albicans ATCC 10231, Candida albicans LM 450, Trichophyton mentagrophytes LM 230, Trichophyton rubrum LM 237, Microsporum canis LM 324.

Results

The hydrodistillation afforded 0.06% (dry weight basis) of pale yellow oil. Thirty-eight compounds representing 90.69% were identified. The major component (36.40%) was found to be (E)-isoelemicin, identified by comparison of its ¹H-NMR experimental data, with literature data. The oil showed good antibacterial activity against Clostridium Sporogenes, Streptococcus mutans and Streptococcus pyogenes with MIC ranging from 13.5 to 126 μg/ml. A weak antifungal activity (MIC 250 μg/ml) was found against Trichophyton rubrum, only.

Conclusions

The antimicrobial activity and the chemical composition of Phyllanthus muellerianus stem bark essential oil are reported for the first time.     

In vitro antimicrobial activity of extracts and compounds of some selected medicinal plants from Cameroon

Aim of the study

Seven extracts and eight compounds from four selected Cameroonian medicinal plants, Solanecio mannii Hook f. (Asteraceae), Monodora myristica Dunal (Annonaceae), Albizia gummifera (J.F. Gmel) C.A. Smith (Fabaceae/Mimosoideae) and Glyphaea brevis (Spreng) Monachino (Tiliaceae), traditionally used for the treatment of hepatitis, parasites and other infectious diseases, were tested in vitro for their antimicrobial activity against Gram-positive (5 species) and Gram-negative (4 species) bacteria species and pathogenic yeasts (2 Candida species), to establish whether or not they have antimicrobial activity and to validate scientifically their use in traditional medicine.

Materials and methods

The agar disc diffusion and the microbroth dilution methods were used to determine the zone of inhibition between the edge of the filter paper and the edge of the inhibition area (IZ) and the minimal inhibitory concentration (MIC) respectively.

Results

The most active extracts against Candida albicans and Candida krusei were respectively the cyclohexane extract from the fruits of Monodora myristica and the ethyl acetate extract from the stem bark of Albizia gummifera (MIC = 6.3 μg/ml for both extracts). The lowest MIC value (1.6 μg/ml) for purified compounds was obtained on Candida albicans with a mixture of linear aliphatic primary alcohols (n-C24H50O to n-C30H62O), with n-hexacosanol (1b) as major compound and mixture of fatty acid esters of diunsaturated linear 1,2-diols (6).

Conclusion

These results afford ground informations for the potential use of the crude extracts of these species as well as of some of the isolated compounds in bacterial and fungal infections.     

Antibacterial,cytotoxic activities and chemical composition of fruits of two Cameroonian Zanthoxylum species

Ethnopharmacological relevance

Zanthoxylum leprieurii (Syn. Fagara leprieurii) is a plant extensively used by traditional healers in Cameroon for the treatment of stomach disorders, gonorrhea, intestinal parasites and sterility. The infusion from the fruit is taken in Cameroon for the treatment of sickle cell anemia, as well. Similarly, the roots of Zanthoxylum zanthoxyloides (Syn. Fagara zanthoxyloides) are also used for wound dressing, sickle cell anemia and as pain reliever, in addition to the uses mentioned for Zanthoxylum leprieurii.

Aim of the study

To scientifically validate their traditional use, the in vitro antimicrobial and anticancer assays were performed. In addition, chemical compositions of their fruit extracts and essential oils were also studied.

Material and methods

The chemical compositions of their essential oils and methanol extracts were studied. GC and GC–MS were used for analyzing the essential oils while various chromatographic methods were used to isolate the pure compounds from methanol extracts. MTT assay was used for the cytotoxic studies of methanol extracts, their fractions and some pure compounds. Antimicrobial activity of methanol extracts, some isolated compounds as well as essential oils was tested against a panel of human pathogenic bacteria by diffusion and dilution methods.

Results

The principal constituents of the essential oil of Zanthoxylum zanthoxyloides were identified as citronellol and geraniol whereas Zanthoxylum leprieurii was rich in E-β-ocimene. Furthermore, we report for the first time, the presence of 7,8-dimethoxycoumarin and sinapic acid in the methanol extract of Zanthoxylum leprieurii. In vitro bioassays reveal that the methanol extracts, essential oil and some of the isolated compounds from fruits of both the plants exhibit moderate anticancer and antimicrobial activities.

Conclusion

Altogether, the present findings justify the traditional use of Zanthoxylum zanthoxyloides and Zanthoxylum leprieurii in the treatment of stomach disorders and cancer disease. The antimicrobial properties in the essential oil of Zanthoxylum zanthoxyloides are of additional significance.     

Chemical and biological characterisation of solvent extracts and essential oils from leaves and fruit of two Australian species of Pittosporum (Pittosporaceae) used in aboriginal medicinal practice

Ethnopharmacological significance

Although no known medicinal use for Pittosporum undulatum Vent. (Pittosporaceae) has been recorded, anecdotal evidence suggests that Australian Aboriginal people used Pittosporum angustifolium Lodd., G. Lodd. & W. Lodd. topically for eczema, pruritis or to induce lactation in mothers following child-birth and internally for coughs, colds or cramps.

Aims of the study

Essential oil composition and bioactivity as well as differential solvent extract antimicrobial activity from Pittosporum angustifolium are investigated here first, to partially describe the composition of volatiles released in traditional applications of Pittosporum angustifolium for colds or as a lactagogue, and second to investigate antibacterial activity related to topical applications. Essential oils were also investigated from Pittosporum undulatum Vent., first to enhance essential oil data produced in previous studies, and second as a comparison to Pittosporum angustifolium.

Materials and methods

Essential oils were hydrodistilled from fruit and leaves of both species using a modified approach to lessen the negative (frothing) effect of saponins. This was achieved by floating pumice or pearlite obsidian over the mixture to crush the suds formed while boiling. Essential oil extracts were analysed using GC–MS, quantified using GC-FID then screened for antimicrobial activity using a micro-titre plate broth dilution assay (MIC). Using dichloromethane, methanol, hexane and H₂O as solvents, extracts were produced from leaves and fruit of Pittosporum angustifolium and screened for antimicrobial activity and qualitative phytochemical character.

Results

Although the essential oil from leaves and fruit of Pittosporum undulatum demonstrated some component variation, the essential oil from fruits of Pittosporum angustifolium had major constituents that strongly varied according to the geographical location of collection, suggesting the existence of at least two chemotypes; one with high abundance of acetic acid decyl ester. This chemotype had high antimicrobial activity whilst the other chemotype had only moderate antimicrobial activity against the three microbial species investigated here. This result may support the occurrence of geographical specificity with regard to ethnopharmacological use. Antimicrobial activity screening of the solvent extracts from Pittosporum angustifolium revealed the leaves to be superior to fruit, with water being the most suitable extraction solvent.

Conclusion

To the best of our knowledge, this study constitutes the first time essential oils, and solvent extracts from the fruits of Pittosporum angustifolium, have been examined employing comprehensive chemical and biological analysis. The essential oil composition presented in this paper, includes components with structural similarity as chemosemiotic compounds involved in mother–infant identification, which may have significance with regard to traditional applications as a lactagogue.     

Antimicrobial activity of the crude extract,fractions and compounds from stem bark of Ficus ovata (Moraceae)

Aim of the study

This study was designed to investigate the antimicrobial activities of the methanol extracts from the stem bark of Ficus ovata (FOB), fractions (FOB1–6) and compounds isolated following bio-guided fractionation [3-friedelanone (1), taraxeryl acetate (2), betulinic acid (3), oleanoïc acid (4), 2-hydroxyisoprunetin (5), 6,7-(2-isopropenyl furo)-5,2,4-trihydroxyisoflavone (6), Cajanin (7) and protocatechuic acid (8)].

Materials and Methods

The micro-dilution method was used for the determination of the minimal inhibition concentration (MIC) and the minimal microbicidal concentration (MMC) against fungi (two species), Gram-positive (three species) and Gram-negative bacteria (five species).

Results

The results of the MIC determinations indicated that the crude extract (FOB), fractions FOB2 and FOB4 as well as compound 5 were active on the entire studied organisms. Other samples showed selective activity, fractions FOB1, FOB3 and FOB5 being active against 50% of the tested microbial species while FOB6 was active on 40%. Compounds 8, 6, 2 and 7 prevented the growth of 80%, 70%, 50% and 20% of the organisms respectively. The lowest MIC value (156g/ml) observed with the crude extract was recorded on Streptococcus faecalis, Candida albicans and Microsporum audouinii. The corresponding value for fractions (39 μg/ml) was noted with FOB4 against Staphylococcus aureus, while that of the tested compounds (10 μg/ml) was observed with compound 8 on Microsporum audouinii. The results of the MMC determination suggested that the cidal effect of most of the tested samples on the studied microorganisms could be expected.

Conclusions

The overall results provided evidence that the studied plant extract, as well as some of the isolated compounds might be potential sources of new antimicrobial drug.     

Davilla elliptica and Davilla nitida: Gastroprotective,anti-inflammatory immunomodulatory and anti-Helicobacter pylori action

Ethnopharmacological relevance

Davilla elliptica and Davilla nitida are species commonly found in the Brazilian Cerrado biome.

Aim of the study

Based on ethnopharmacological and phytochemical analyses, methanolic extracts from leaves of Davilla elliptica (EDE) and Davilla nitida (EDN) were evaluated for their anti-ulcer, anti-inflammatory, immunological and anti-Helicobacter pylori activities.

Materials and methods

The gastroprotective action of both extracts was evaluated in rodent experimental models (HCl/ethanol, ethanol or NSAID). We also evaluated anti-inflammatory (carrageenin-induced rat hind paw edema), immunomodulatory (murine peritoneal macrophages) and antibacterial action of both extracts against a standard strain of Helicobacter pylori.

Results

EDE and EDN (500 mg/kg) were able to protect gastric mucosa against HCl/ethanol solution (EDE 63%; EDN 59%), absolute ethanol (EDE 95%; EDN 88%), and also against injurious effect of NSAID (EDE 77%; EDN 67%). When EDE and EDN were challenged with sulfhydryl depleter compound, the gastroprotective action of both extracts was completely abolished. EDE had gastroprotective effect related to increase of glutathione bioavailability and stimulated higher levels of NO, H₂O₂ and TNF-α production. Otherwise EDN showed better anti-Helicobacter pylori action than EDE. Neither extracts presented anti-inflammatory activity by oral route.

Conclusion

The phytochemical investigation showed that both extracts possess phenolic acid derivatives, acylglycoflavonoids and condensed tannins with evident quantitative variations that probably influenced the pharmacological differences between extracts.     

Assessment of the genotoxic and mutagenic properties of Himatanthus articulatus bark extracts used as phytotherapeutic drug in the Amazon

Ethnopharmacological relevance

Himatanthus articulatus (Apocynaceae) is a plant native to the Amazon, popularly used to treat external ulcers, tumors, inflammations, cancer, syphilis and malaria.

Aim of the study

To investigate the in vivo genotoxic and mutagenic potential of this plant, using the comet assay and the micronucleus test.

Material and methods

Female and male adult mice were treated with 500 mg/kg, 1000 mg/kg or 2000 mg/kg of Himatanthus articulatus aqueous or ethanolic bark extracts by gavage for two consecutive days. In addition, blood slides were exposed to hydrogen peroxide (ex vivo) to evaluate the anticlastogenic effect using the comet assay. The HPLC analyses indicated plumieride as the main constituent of both extracts from Himatanthus articulatus barks.

Results

No differences between genders were observed. Micronuclei were observed only in the group treated with the highest dose of both extracts. Conversely, lower doses of these extracts showed protective effects to DNA against damage induced by hydrogen peroxide, indicating an important antigenotoxic effect.

Conclusions

The toxicological evaluation indicated that the extracts are non-genotoxic and reduce the clastogenic damage induced by hydrogen peroxide. In part, this result can be atributted to the phytochemical profile of Himatanthus articulatus, which presents iridoids and phenolic compounds.     

Effect of Tectona grandis Linn. on dexamethasone-induced insulin resistance in mice

Ethnopharmacological relevance

The bark of Tectona grandis Linn. is traditionally used in the treatment of diabetes.

Aim

The present study was undertaken to investigate the effect of ethanolic extract of bark of Tectona grandis Linn. (TG) in dexamethasone-induced insulin resistance in mice.

Materials and methods

Mice were treated with prestandardised dose of dexamethasone for 22 days and effect of TG at the doses of 50, 100 and 200 mg/kg, p.o. on plasma blood glucose level, serum triglyceride level, glucose uptake in skeletal muscle, levels of hepatic antioxidant enzymes (GSH, SOD, catalase and LPO), and body weight were observed.

Results

TG showed significant decrease in plasma glucose and serum triglyceride levels (p < 0.01) at the dose of 100 and 200 mg/kg, p.o. and also stimulated glucose uptake in skeletal muscle. The levels of antioxidant enzymes GSH, SOD, and catalase were significantly increased (p < 0.01) and there was significant decrease (p < 0.01) in level of LPO.

Conclusion

Hence it can be concluded that Tectona grandis may prove to be effective in the treatment of Type-II Diabetes mellitus owing to its ability to decrease insulin resistance.     

An antimicrobial evaluation of plants used for the treatment of respiratory infections in rural Maputaland,KwaZulu-Natal,South Africa

Ethnopharmacological relevance

Abundant availability of medicinal plants in the study area offers low cost health care, but scientific validation is needed in order to lend credibility to the traditional use against respiratory infections.

Aim of the study

This study focussed on determining the antimicrobial efficacies of 30 plant species (independently and in various combinations) used for respiratory related infections in rural Maputaland.

Materials and methods

In vitro minimum inhibitory concentration (MIC) assays were undertaken on dichloromethane-methanol (CH₂Cl₂: MeOH) and aqueous extracts, as well as the hydro-distilled essential oils (for aromatic plants). Selected plant parts were assessed for antimicrobial activity against a range of respiratory pathogens i.e. Cryptococcus neoformans (ATCC 14116), Klebsiella pneumoniae (ATCC 13883), Moraxella catarrhalis (ATCC 23246), Mycobacterium smegmatis (ATCC 14468) and Staphylococcus aureus (ATCC 6538). The sum of the fractional inhibitory concentrations (∑FIC) was determined for plants traditionally used in combination. Isobolograms represent MIC values for a selection of interactions where two plants were combined in various ratios.

Results

The most antimicrobially active aqueous extracts were that of Ozoroa obovata and Sclerocarya birrea (0.10 mg/ml) while organic extracts from Parinari capensis subsp. incohata and Tetradenia riparia demonstrated the most noteworthy (0.03 mg/ml) activity. Although both Lippia javanica and Eucalyptus grandis were by far the most popular plants traditionally used for respiratory infections, the antimicrobial activity was mostly only moderate. Furthermore, the traditional use in a 1:1 combination did not display strong antimicrobial interactions, but isobolograms demonstrate (against some test organisms) that when combined in various ratios, predominant additive interactions are evident where E. grandis was present in larger proportions. The combination of E. grandis with O. obovata demonstrated synergism against both C. neoformans and K. pneumoniae, with ∑FIC values of 0.34 and 0.28 respectively. Various ratios of these two plants also demonstrated a predominantly synergistic profile.

Conclusion

Although this in vitro study supports the traditional use of some plants independently and in combination for the treatment of respiratory ailments in rural Maputaland, results demonstrate that the traditional selection of plants in higher frequency do not necessarily correlate with higher antimicrobial efficacy.     

Antimicrobial interactions of Khoi-San poly-herbal remedies with emphasis on the combination; Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus

Ethnopharmacological relevance

Plants are often combined in traditional herbal remedies to increase medicinal efficacy, thus this investigation provides some insight into the antimicrobial efficacies of selected combinations.

Aims of the study

The first aim was to scientifically validate antibacterial efficacy of plant mixtures that are traded within peri-urban centres of Cape Town (Western Cape, South Africa). This was followed by an in-depth evaluation of the most antimicrobially active mixture; Agathosma crenulata, Dodonaea viscosa with Eucalyptus globulus.

Materials and methods

Methanol and aqueous extracts of six plant mixtures were screened for antibacterial properties against two Gram-negative and two Gram-positive bacteria using the minimum inhibitory microdilution method. Thereafter, chloroform: methanol (1:1; v/v) extracts, essential oils and aqueous extracts of Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus were assayed for antimicrobial activity independently and in various combinations. The fractional inhibitory concentration indices (?FIC) were determined for double and triple plant combinations to establish antimicrobial interactions.

Results

From the six plant mixtures prepared by herbalists, a methanol extract derived from combining Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus showed the best antibacterial activity. The MIC values of 49 μg/ml for Staphylococcus aureus and Bacillus subtilis, and 98 μg/ml for Klebsiella pneumoniae and Escherichia coli were recorded. When Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus were mixed in various 1:1 combinations, mostly additive and synergistic interactions were noted. The most noteworthy synergistic (ΣFIC value 0.07) 1:1 combinations were observed for the chloroform: methanol extracts of Agathosma crenulata mixed with Eucalyptus globulus against Klebsiella pneumoniae, Staphylococcus aureus and Bacillus subtilis. When combined in a mixture of three plants (1:1:1), enhanced efficacy was evident against most of the pathogens, for both organic and aqueous extracts. The triple combination against Bacillus subtilis demonstrated the greatest synergy (ΣFIC values of 0.03).

Conclusions

The enhanced antimicrobial efficacy and synergistic interactions noted for some of the mixtures, particularly the combination of Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus support the Western Cape Khoi-San traditional medicinal practices of combining plants for enhanced efficacy.     

Alpha-amylase inhibitory activity of two Anthocleista species and in vivo rat model anti-diabetic activities of Anthocleista djalonensis extracts and fractions

Ethnopharmacological relevance

Anthocleista djalonensis (A. Chev) and Anthocleista vogelii Planch are plants being used in West Africa traditionally to treat various diseases such as malaria, hernia, hypertension, stomach aches, hemorrhoids, syphilis, and diabetes. Diabetes causes about 5% of all deaths globally each year. Chemotherapeutic agents such as biguanides, sulfonylureas, and thiozolidinediones are available for the treatment of diabetes, however, they have undesirable side effects. The need for newer, more effective and less toxic drugs is imperative and the biodiversity of Nigeria has a high potential for drug discovery based on plants used in the ethnomedicine.

Aim of the study

To investigate the leaves, stem bark and roots of these plants for their probable alpha-amylase inhibitory activities and establish their anti-diabetic activities. The overall goal is do bioassay-guided fractionation and isolation of active anti-diabetic compounds.

Materials and methods

Powdered samples (leaves, stem bark and roots) macerated with 80% aqueous methanol were evaluated in vitro using alpha-amylase inhibitory assay while in vivo investigations were carried out on hyperglycemic rats. Diabetes was induced in albino rats by an intraperitoneal injection of alloxan monohydrate (80 mg/kg). Plant extracts (1 g/kg) were given orally for 7 days, while blood glucose levels were monitored using a one touch glucometer. The crude methanol extracts found to be most active were further partitioned into hexane and ethyl acetate fractions which were also tested in vivo on the diabetic animals.

Results

The leaves and stem bark crude methanol extracts of Anthocleista djalonensis gave comparable α-amylase inhibition of 73.66% and 72.90%, respectively which were quite higher than the 38.93% and 22.90% of the same plant parts given by Anthocleista vogelii. The crude stem bark extract of Anthocleista djalonensis exhibited significant peak blood glucose reduction on day 6 (72.59%, p<0.05) which was higher than the leaves or roots which gave 45.73% and 47.46% (p<0.05), respectively The stem bark ethyl acetate fraction of Anthocleista djalonensis gave reduction in blood glucose level of 60.86% (p<0.05).

Conclusion

From our results, the leaves, stem bark and whole root of both plants exhibited α-amylase inhibitory activities with Anthocleista djalonensis also showing good anti-diabetic activities in vivo indicating that they contain active principles for the management of diabetes. There is justification for the use of the plants traditionally to manage diabetes.     

Validation of smoke inhalation therapy to treat microbial infections

Aim of the study

In traditional healing, the burning of selected indigenous medicinal plants and the inhalation of the liberated smoke are widely accepted and a practiced route of administration. This study elucidated the rationale behind this commonly practiced treatment by examining the antimicrobial activity for five indigenous South African medicinal plants commonly administered through inhalation (Artemisia afra, Heteropyxis natalensis, Myrothamnus flabellifolius, Pellaea calomelanos and Tarchonanthus camphoratus).

Material and Methods

An apparatus was designed to simulate the burning process that occurs in a traditional setting and the smoke fraction was captured for analysis and bioassay. Methanol and acetone extracts as well as the essential oil (for the aromatic species) were prepared and assayed in parallel with the smoke fraction.

Results

Antimicrobial data revealed that in most cases, the ‘smoke-extract’ obtained after burning had lower minimum inhibitory concentration (MIC) values than the corresponding solvent extracts and essential oils. The combustion, acetone and methanol extracts produced different chromatographic profiles as demonstrated for Pellaea calomelanos where several compounds noted in the smoke fraction were not present in the other extracts.

Conclusion

These results suggest that the combustion process produces an ‘extract’ with superior antimicrobial activity and provides in vitro evidence for inhalation of medicinal smoke as an efficient mode of administration in traditional healing.     

Antimicrobial phenylpropanoids from the Argentinean highland plant Parastrephia lucida (Meyen) Cabrera

Ethnopharmacological relevance

The Argentinean highland plant Parastrephia lucida (Meyen) Cabrera is used in traditional medicine as an antiseptic and anti-inflammatory crude drug.

Aim of the study

To relate the antimicrobial effect of the crude drug with the constituents of the active fractions and traditional use.

Materials and methods

Assay-guided isolation of the methanol (MeOH) plant extract was carried out using bacteria and yeasts as target organisms. Both ATCC and local strains were included in the study. The antimicrobial fractions and compounds were detected by bioautographic assays. Minimum inhibitory concentrations (MIC) of each extract and fraction were determined and compared with reference antibiotics. Fractions were analyzed by HPLC–DAD, GC–MS, ¹H NMR and ¹³C NMR.

Results

From the MeOH extract of the plant, assay-guided isolation of the antimicrobial constituents led to 12 phenylpropanoids and two simple phenolics. Most of the compounds occurring in the active fractions were E-caffeoyl or E-cinnamoyl esters including prenyl and phenethyl derivatives. The MIC values of the most active fractions ranged between 12.5 and 200 μg/mL against reference strains and local isolates of Staphylococcus aureus and Enterococcus faecalis.

Conclusions

The antimicrobial effect found in the crude drug was associated with mixtures of phenylpropanoids, including prenyl and phenethyl esters of caffeic and cinnamic acids. The results support at least in part the traditional use of the plant as local antiseptic.