Bacterial communities associated with Acrobeles complexus nematodes recovered from tomato crops in South Africa.

The productivity of agricultural ecosystems is heavily influenced by soil-dwelling organisms. To optimize agricultural practices and management, it is critical to know the composition, abundance, and interactions of soil microorganisms. Our study focused on Acrobeles complexus nematodes collected from tomato fields in South Africa and analyzed their associated bacterial communities utilizing metabarcoding analysis. Our findings revealed that A. complexus forms associations with a wide range of bacterial species. Among the most abundant species identified, we found Dechloromonas sp., a bacterial species commonly found in aquatic sediments, Acidovorax temperans, a bacterial species commonly found in activated sludge, and Lactobacillus ruminis, a commensal motile lactic acid bacterium that inhabits the intestinal tracts of humans and animals. Through principal component analysis (PCA), we found that the abundance of A. complexus in the soil is negatively correlated with clay content (r = -0.990) and soil phosphate levels (r = -0.969) and positively correlated with soil sand content (r = 0.763). This study sheds light on the bacterial species associated to free-living nematodes in tomato crops in South Africa and highlights the occurrence of various potentially damaging and beneficial nematode-associated bacteria, which can in turn, impact soil health and tomato production.

Evaluation of the Antioxidant, Cytotoxicity, Antibacterial, Anti-Motility, and Anti-Biofilm Effects of Myrothamnus flabellifolius Welw. Leaves and Stem Defatted Subfractions.

The formation of biofilms underscores the challenge of treating bacterial infections. The study aimed to assess the antioxidant, cytotoxicity, antibacterial, anti-motility, and anti-biofilm effects of defatted fractions from Myrothamnus flabellifolius (resurrection plant). Antioxidant activity was assessed using DPPH radical scavenging and hydrogen peroxide assays. Cytotoxicity was screened using a brine shrimp lethality assay. Antibacterial activity was determined using the micro-dilution and growth curve assays. Antibiofilm potential was screened using the crystal violet and tetrazolium reduction assay. Liquid-liquid extraction of crude extracts concentrated polyphenols in the ethyl acetate and n-butanol fractions. Subsequently, these fractions had notable antioxidant activity and demonstrated broad-spectrum antibacterial activity against selected Gram-negative and Gram-positive bacteria and Mycobacterium smegmatis (MIC values < 630 µg/mL). Growth curves showed that the bacteriostatic inhibition by the ethyl acetate fractions was through the extension of the lag phase and/or suppression of the growth rate. The sub-inhibitory concentrations of the ethyl acetate fractions inhibited the swarming motility of Pseudomonas aeruginosa and Klebsiella pneumoniae by 100% and eradicated more than 50% of P. aeruginosa biofilm biomass. The polyphenolic content of M. flabellifolius plays an important role in its antibacterial, anti-motility, and antibiofilm activity, thus offering an additional strategy to treat biofilm-associated infections.

Antimicrobial, antioxidant and cytotoxic activities of the leaf and stem extracts of Carissa bispinosa used for dental health care.

BACKGROUND: Carissa bispinosa (L.) Desf. ex Brenan is one of the plants used traditionally to treat oral infections. However, there is limited data validating its therapeutic properties and photochemistry. The aim of this study was to investigate the protective efficacy of the leaf and stem extracts of C. bispinosa against oral infections. METHODS: The phenolic and tannin contents were measured using Folin-Ciocalteau method after extracting with different solvents. The minimum inhibitory concentrations (MIC) of the extracts were assessed using the microdilution method against fungal (Candida albicans and Candida glabrata) and bacterial (Streptococcus pyogenes, Staphylococcus aureus and Enterococcus faecalis) strains. The 2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing power (FRP) models were utilised to assess the antioxidant potential of the extracts. Cytotoxicity of the leaf acetone extract was evaluated using the methylthiazol tetrazolium assay. RESULTS: The methanol leaf extract had the highest phenolic content (113.20 mg TAE/g), whereas hexane extract displayed the highest tannin composition of 22.98 mg GAE/g. The acetone stem extract had the highest phenolic content (338 mg TAE/g) and the stem extract yielded the highest total tannin content (49.87 mg GAE/g). The methanol leaf extract demonstrated the lowest MIC value (0.31 mg/mL), whereas the stem ethanol extract had the least MIC value of 0.31 mg/mL. The stem methanol extract had the best DPPH free radical scavenging activity (IC50, 72 µg/mL) whereas the stem ethanol extract displayed maximum FRP with absorbance of 1.916. The leaf acetone extract had minimum cytotoxicity with the lethal concentration (LC50) of 0.63 mg/mL. CONCLUSIONS: The results obtained in this study validated the protective effect of C. bispinosa against oral infections.

Phytochemical Content, Antibacterial Activity, and Antioxidant, Anti-Inflammatory, and Cytotoxic Effects of Traditional Medicinal Plants against Respiratory Tract Bacterial Pathogens.

Respiratory tract infections (RTIs) are frequent ailments among humans and are a high burden on public health. This study aimed to determine the in vitro antibacterial, anti-inflammatory, and cytotoxic effects of indigenous medicinal plants used in the treatment of RTIs, namely, Senna petersiana, Gardenia volkensii, Acacia senegal, and Clerodendrum glabrum. Dried leaves were extracted using various organic solvents. Antibacterial activity was quantified using the microbroth dilution assay. Protein denaturation assays were used to evaluate anti-inflammatory activity. The cytotoxicity of the extracts towards THP-1 macrophages was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Antioxidant activity was determined using free radical scavenging activity and ferric-reducing power. Total polyphenolics were quantified. Liquid chromatography mass spectrometry was used to evaluate the acetone plant extracts. Nonpolar extracts had noteworthy antibacterial activity against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Mycobacterium smegmatis where MIC values ranged between 0.16 and 0.63 mg/mL. At 100 µg/mL, A. senegal, G. volkensii, and S. petersiana had a nonsignificant effect on the viability of the THP-1 macrophages. The LC-MS analysis of the leaf extracts of S. petersiana detected Columnidin, Hercynine, L-Lysine citrate, and Gamma-Linolenate. A pentacyclic triterpenoid, cochalate, was detected in G. volkensii. Two flavonoids 7-hydroxy-2-(4-methoxyphenyl)-4-oxo-chroman-5-olate and (3R)-3-(2,4-dimethoxyphenyl)-7-hydroxy-4-oxo-chroman-5-olate were detected in the C. glabrum extract. The findings from this study indicated that the leaves of the selected plant extracts possess antioxidant, anti-inflammatory, and antibacterial activity. Therefore, they may serve as good candidates for further pharmaceutical investigations.

Chemical Profile, Antioxidant and Antibacterial Activities, Mechanisms of Action of the Leaf Extract of Aloe arborescens Mill.

Aloe arborescens Mill's extracts have been explored for antibacterial and antioxidant efficacies. However, there is limited information on its chemical composition and mechanism of action. The purpose of this study was to assess the chemical composition, antibacterial and antioxidant activities and mechanism of the whole leaf extract of A. arborescens Mill. The phytochemical profile was analysed with gas chromatography mass spectrometry (GC-MS). The antioxidant and antibacterial activities were screened using 1,1diphenyl2picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and micro-dilution assays, respectively. The effects of the extract on the bacterial respiratory chain dehydrogenase, membrane integrity and permeability were analysed using iodonitrotetrazolium chloride, 260 absorbing materials and relative electrical conductivity assays. GC-MS spectrum revealed 26 compounds with N,N'-trimethyleneurea (10.56%), xanthine (8.57%) and 4-hexyl-1-(7-ethoxycarbonylheptyl)bicyclo[4.4.0]deca-2,5,7-triene (7.10%), being the major components. The extract also exhibited antioxidant activity with median concentration (IC50) values of 0.65 mg/mL on DPPH and 0.052 mg/mL on ABTS. The extract exhibited minimum inhibitory concentration (MIC) values ranging from 0.07 to 1.13 mg/mL. The extract inhibited the bacterial growth by destructing the activity of the respiratory chain dehydrogenase, membrane integrity and permeability. Therefore, the leaf extract has the potential to serve as a source of antibacterial and antioxidant compounds.

Bioprospecting of endophytic actinobacterium associated with Aloe ferox mill for antibacterial activity.

BACKGROUND: The emergence of drug resistance among pathogens has resulted in renewed interest in bioprospecting for natural microbial products. METHODS: This study aimed to bioprospecting endophytic actinobacterium associated with Aloe ferox Mill for its antibacterial activity. Endophytic actinomycetes were isolated from the gel of A. ferox Mill by surface sterilization technique using actinomycete isolation agar. The isolate with a promising antibacterial activity was identified using 16S rRNA sequence analysis. The minimum inhibitory concentration (MIC) of the extract was assessed by the micro-dilution method and its effect on the respiratory chain dehydrogenase (RCD) activity was ascertained by the iodonitrotetrazolium chloride (INT) assay. Fourier transform-infrared spectrophotometer (FTIR) and gas chromatography-mass spectrophotometry (GC-MS) were employed to identify functional groups and the chemical constituents, respectively. RESULTS: The actinobacterium was found to be Streptomyces olivaceus CP016795.1. Its extract displayed noteworthy antibacterial activity (MIC ≤1 mg/mL) against Staphylococcus aureus (ATCC 25925), Bacillus cereus (ATCC 10102), and Escherichia coli (ATCC 25922); and showed an inhibitory effect on the RCD activity. FTIR spectrum displayed hydroxyl, amine, and aromatic groups, and the GC-MS revealed 5-Hydroxymethylfurfural as the main constituent (19.47%). CONCLUSIONS: S. olivaceus CP016795.1 can serve as a potential source of effective antibacterial compounds.

Isolation and Optimisation of Culture Conditions for a Marine Bioflocculant-Producing Bacterium and Application of Its Bioflocculant in Wastewater Treatment.

The application of bioflocculants has become an alternative to that of chemical flocculants in wastewater treatment due to their environmental friendliness and non-toxic effects. This study aimed at isolating a bioflocculant-producing bacterium from marine water, optimisation of its culture conditions, and investigation of the removal efficiency of its bioflocculant on pollutants in wastewater. The bacterium was identified by 16S rRNA gene analysis. Optimal carbon and nitrogen sources, inoculum size, temperature, pH, and time were determined by the one-factor-at-a-time assay. The cytotoxicity of the bioflocculant was assessed on African green monkey kidney and bovine dermis cells using a tetrazolium-based columetric (MTT) method. Its removal efficiencies on chemical oxygen demand (COD), biological oxygen demand (BOD) and sulphur were determined using the Jar test method. The bacterial isolate was identified as Ochrobactrum oryzae AB84113. A maximum flocculating activity of 92% and a yield of 3.768 g/L were obtained when a 1% (v/v) inoculum size was used in the presence of starch and yeast extract at pH 7, 30 °C, and after 72 h of cultivation. The bioflocculant demonstrated non-cytotoxic effects on bovine dermis and African green monkey kidney cells. The bioflocculant removed 98% COD, 91% BOD and 86% of Sulphur. The bioflocculant has potential for pollutant removal from industrial wastewater.

Exploring the anti-HIV-1 reverse transcriptase, anti-inflammatory, anti-cancer activities and cytotoxicity of two fermented commercial herbal concoctions sold in Limpopo Province of South Africa.

BACKGROUND AND OBJECTIVES: The use of herbal concoctions is very popular in South Africa, including Limpopo Province. The herbal concoctions are claimed to be capable of treating numerous illnesses such as ulcers, cancer, HIV/AIDS, diabetes, certain STDs, blood cleansing to mention but a few. The focus of this study was to evaluate the anti-HIV 1 reverse transcriptase, anti-inflammatory and anti-cancerous activities as well as cytotoxic effects of 2 fermented herbal concoctions used for the treatment of the related ailments in Limpopo province of South Africa. METHOD: Two fermented herbal concoctions obtained from a herbalist in Polokwane were extracted with 80% acetone. The anti-HIV activity of the herbal concoctions was determined using the anti-HIV reverse transcriptase assay. The anti-cancer and cytotoxic effects of the herbal concoctions were evaluated using cancerous Human Colon (HT-29) cells and the normal human Hepatoma cells (C3A) respectively. RESULTS: Notable anti-HIV reverse transcriptase activity was observed from the 80% acetone fraction of herbal concoction 1 (IC50 38.031 µg/mL) which exhibited better activity than the positive control Lamivudine (IC50 40.90 µg/mL). There was variation in the anti-inflammation activity as determined by the sPL2, 15-LOX and COX enzyme assays. The only concerning matter was the high COX-1 activity in some of the extracts, which is not desirable due to the mucosal protection action of COX-1 enzyme. The herbal concoctions did not exhibit cytotoxic effects on normal human cells, however, toxicity against cancerous cells was observed. CONCLUSION: The herbal concoctions displayed some considerable pharmacological effects against various ailments as claimed by the herbalist. More work to ascertain the toxicity of both concoctions against cancerous cells need to be followed as this could lead to the discovery of anticancer drugs.

Phytochemical Investigation, Antioxidant and Antimycobacterial Activities of Schkuhria pinnata (Lam) Thell Extracts Against Mycobacterium smegmatis.

The focus of this study was to evaluate the antioxidants and antimycobacterial activities of extracts of Schkuhria pinnata. Serial exhaustive extraction procedure was employed using solvents of varying polarity to obtain the desired extracts. Thin layer chromatography and standard chemical tests were used to analyze phytochemicals constituents. Free radical scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH) methods were used to detect the presence of antioxidant compounds. Antimycobacterial activity was evaluated using microdilution and bioautography assays. A variety of secondary metabolites such as flavonoids, tannins, and alkaloids were detected in the extract. Ethyl acetate and acetone extracts had high antioxidant activity on chromatograms eluted in ethyl acetate/methanol/water while methanol extract at various concentrations had the best scavenging activity. The minimum inhibitory concentration (MIC) values ranged from 0.02 to 2.50 mg/mL. Total phenol content was 55.33 ± 3.51 mg of gallic acid equivalent (GAE)/g and higher when compared with flavonoids (4.00 ± 0.35 mg of quercetin equivalent [QE]/mg) and tannin content (28.00 ± 1.73 mg of GAE/g). The most effective antimycobacterial activity against Mycobacterium smegmatis was observed with the lowest inhibitory concentrations of acetone (0.27 mg/mL), dichloromethane (0.32 mg/mL), and ethyl acetate (0.32 mg/mL) in that order. In massive extraction, hexane and dichloromethane had the greatest inhibitory bands on benzene/ethanol/ammonium hydroxide bioautograms. Antimmycobacterial activity gives promising potential leads of S pinnata extracts to be used in the development of antimycobacterial drugs. The presence of antioxidant and antimycobacterial compounds requires further isolation and purification.

Antibacterial interactions, anti-inflammatory and cytotoxic effects of four medicinal plant species.

BACKGROUND: The constant emergence of antibiotic resistant species and the adverse side effects of synthetic drugs are threatening the efficacy of the drugs that are currently in use. This study was aimed at investigating the possible antibacterial interactions, anti-inflammatory and cytotoxic effects of selected medicinal plants based on their traditional usage. METHODS: The acetone extracts of four plant species were assessed independently and in combination for antibacterial activity using microdilution assay and the sum of the fractional inhibitory concentration (FIC) was calculated. The ability of Dombeya rotundifolia and Schkuhria pinnata extracts to inhibit the production of reactive oxygen species (ROS) in LPS induced RAW 264.7 macrophage cells was evaluated using Dichloro-dihydro-fluorescein diacetate (H2DCF-DA) assay to determine anti-inflammatory potential and the toxicity on African green monkey kidney (Vero) cells was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The antibacterial efficacies of the different combinations of Schkuhria pinnata (A), Commelina africana (B), Dombeya rotundifolia (C) and Elephantorrhiza elephantina (D) plants varied from combination to combination. Synergistic effects were only exhibited against P. aeruginosa, while the antagonistic effects were only observed against E. coli. Both S. pinnata and D. rotundifolia demonstrated anti-inflammatory potential by inhibiting the production of ROS in a dose dependant manner. The cytotoxicity of the plants (LC50 values) ranged from < 25.0 to 466.1 µg/mL. S pinnata extract was the most toxic with the lowest LC50 value of < 25.0 µg/mL. CONCLUSIONS: The synergistic interaction observed indicates that combinational therapy may improve biological activity. This report highlights the anti-inflammatory potential of S. pinnata and D. rotundifolia; which could be exploited in the search for anti-inflammatory agents. However, the cytotoxicity of S. pinnata highlights the importance of using this plant with caution.

Antibacterial activity and in situ efficacy of Bidens pilosa Linn and Dichrostachys cinerea Wight et Arn extracts against common diarrhoea-causing waterborne bacteria.

BACKGROUND: Bidens pilosa and Dichrostachys cinerea extracts were investigated for the antibacterial properties against waterborne diarrhoeagenic bacteria. METHODS: The plant materials were extracted using the direct and serial exhaustive methods using solvents of varying polarities, namely, hexane, dichloromethane, ethyl acetate, acetone and methanol. Qualitative phytochemical analysis and quantitative determination of total phenolic content of the leaf powders of the two plants were tested. The antioxidant activities of the plants were determined using the 2, 2-diphenyl-1-picrylhydrazyl method. The toxic effect of the extracts on C2C12 muscle cell line were assessed by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide method and the antibacterial activity was determined using the serial microbroth dilution. RESULTS: Methanol leaf extracts both plants had the highest yield in both direct and serial exhaustive extraction methods. Phytochemical profiling of the extracts displayed the presence of various secondary metabolites. The Benzene: ethanol: ammonia hydroxide solvent system showed a good resolution of chemical compounds in plant extracts from both plants. Most antioxidant compounds observed were developed in chloroform: ethyl acetate: formic acid and ethyl acetate: methanol: water solvent systems. All the bacterial species tested were sensitive to the effect of different extracts of both plant species, with E. coli being less sensitive to the effect of the extracts from D. cinerea. Following the simulated gastric fluid (SGF) treatment, a decrease in the antibacterial potency of the extracts was observed. No extract was toxic to the C2C12 muscle cell line. CONCLUSION: The presence of the secondary metabolites and nontoxic effect of the two plants tested may affirm the medicinal value of these leaf extracts. Our results suggest that B. pilosa and D. cinerea contain constituents with antioxidant and antimicrobial activities, which could be used in the treatment of diarrhoea in a case where untreated surface water is used.

In Vitro Assessment of Selected Medicinal Plants Used by the Bapedi Community in South Africa for Treatment of Bacterial Infections.

There is very little scientific information on indigenous plants used for medicinal purposes. Therefore, the aim of the study was to evaluate the antibacterial and antioxidant activities of the leaves of Commelina africana, Elephantorrhiza elephantina, Dombeya rotundifolia, and the whole plant excluding the roots of Schkuhria pinnata indigenous medicinal plants from the Limpopo Province, which may be used for the treatment in humans infected with bacterial pathogens. The screening of different phytoconstituents using standard methods revealed the presence of terpenoids, flavonoids, tannins, and saponins. The total phenolic, tannin, and flavonoid content were estimated using spectrophotometric methods. D rotundifolia had the highest amounts of phenol (259.00 ± 2.65 mg of gallic acid equivalent [GAE]/g), tannin (330.33 ± 15.63 mg of GAE/g), and flavonoid (19.90 ± 0.75 mg of quercetin equivalent [QE]/g) content, while S pinnata had the least. All the plants had antibacterial activity against Escherichia coli and Pseudomonas aeruginosa with the bioautography method. The microbroth dilution assay revealed antibacterial activity with minimum inhibitory concentration values raining from 0.02 to 2.50 mg/mL. D rotundifolia had the highest antioxidant activity in both the free radical scavenging and ferric reducing power methods. S pinnata and C africana extracts had high antibacterial activity, while D rotundifolia had high antioxidant activity, which made them potential plants for isolation of bioactive compounds and possible application in the pharmaceutical industry.

Phytochemical Analysis, Antioxidant and Antibacterial Properties of Spilanthes mauritiana Used Traditionally in Limpopo Province, South Africa.

Spilanthes mauritiana belonging to the family Asteraceae, was screened for biological activity against bacterial pathogen. Antibacterial activity of the plant was investigated using microbroth dilution assay and bioautography. Total phenols and tannins of the extract were 52.47 ± 2.29 and 23.9 ± 1.18 as mg of gallic acid equivalents, respectively. Total flavonoid content was 25.1 ± 0.79 as mg of quercetin equivalents. Free radical scavenging activity of constituents in all the extract, against 2,2-diphenyl-1-picrylhydrazyl showed minimal activity. All extracts contained compounds with antibacterial activity against tested pathogens. Hexane extract had an average minimum inhibitory concentration value of 2.50 mg/mL, followed by methanol extract (1.72 mg/mL), acetone and dichloromethane extracts (1.96 mg/mL). The present study revealed the presence of compounds in S mauritiana with potent antibacterial activity against tested pathogens that are exhibiting the observed activity independent of other constituents contained in the extracts.

Evaluation of Herbal Concoctions Sold at Ga Maja (Limpopo Province) in South Africa and In Vitro Pharmacological Evaluation of Plants Used to Manufacture the Concoctions.

The aim of the study was to evaluate the biological activities and safety of commercial herbal concoctions manufactured in Ga Maja (Limpopo province). Microbial contamination was evaluated by spread-plating the concoctions on agar plates. The VITEK 2 instrument was used for identification of the pure cultures. Nutritional content of the concoctions was determined. Thin layer chromatography was used to analyze the chemical constituents of the extracts. The microdilution assay and bioautography were used to evaluate antimicrobial activity against selected microorganisms. Sodium, potassium, and zinc were elements most abundant in the concoctions. Phytochemical screening revealed the presence of various phytoconstituents. Acetone extracts of Hypoxis hemerocallidea and Kirkia wilmsii extracts had antioxidant activity. The minimum inhibitory concentrations values against test bacteria ranged between 0.02 and 0.63 mg/mL. Further studies are required to isolate bioactive compounds and evaluate their cytotoxicity. Caution in the consumption of the herbal mixtures should be adhered to.

Anti-mycobacteria potential and synergistic effects of combined crude extracts of selected medicinal plants used by Bapedi traditional healers to treat tuberculosis related symptoms in Limpopo Province, South Africa.

BACKGROUND: Tuberculosis is an infectious communicable disease and the causative agent of the disease has over the years developed resistance to streamline chemotherapeutic agents with dire consequences and there is a need for development of new and more potent alternatives. METHODS: Constituents of leaves material of Combretum heroroense, Citrus lemon and Apodytes dimidiata were serially extracted using solvents of varying polarity. TLC finger print profile of the different extracts were determined by spraying eluted plates with vanillin sulphuric acid and 2, 2- diphenylpicryl hydrazyl (DPPH) for the presence of antioxidant constituents. Presence of different phytochemicals was determined using standard chemical test. Bioautography was used to determine the number of compounds present in sub-fractions active against Mycobacterium smegmatis. Minimum inhibitory concentration (MIC) values extract and sub-fractions were determined using serial microplate dilution method against M. smegmatis (ATCC 1441), M. tuberculosis (ATCC H37Rv) and multi-drug resistant TB (MDR-TB) field strain. Synergy of the crude extracts of the three plants was determined using microplate dilution method against M. smegmatis. RESULTS: Mass extracted by different solvents was less than 6% dry weight for all the plants. Phlobatannins were not detected in A. dimidiata, C. heroroense and C. lemon as well as cardiac glycosides in C. lemon and A. dimidiata, and saponins in C. heroroense. Sub-fractions of the different plants were shown to contain constituents with antioxidant activity with the highest number detected in C. heroroense. Bioautography results reveal the presence of a compound(s) in the ethyle acetate sub-fraction of C. heroroense and butanol, methanol/water, ethyl acetate and water no.2 subfractions of A. dimidiata, active against M. smegmatis that were not shown to have antioxidant capacity. MIC results for different crude extracts of the three plants against M. smegmatis ranges from 0.1 to 3 mg/ml. The average MIC for the synergistic effect of the plants ranged from 0.04 mg/ml to 1.25 mg/ml. An activity greater than that obtained for the reference drugs was shown for the butanol and hexane fractions of A. dimidiata (0.47 mg/ml) against the field strain of MDR-TB while that obtained for the M.TB (ATCC H37Rv) was 0.31 mg/ml. CONCLUSION: A significant finding shown in this study reveals the potent anti-mycobacteria potential of sub-fractions of A. dimidiata against MDR-TB field strain that can lead to the isolation of compounds that can be used to counter resistant strains of tuberculosis.

Isolation of alpha-linolenic acid from Sutherlandia frutescens and its inhibition of Mycobacterium tuberculosis' shikimate kinase enzyme.

BACKGROUND: Sutherlandia frutescens (L) R.Br. is one of traditional herbal medicines that formed the basis of primary health care systems since the earliest days and is still widely used. Sutherlandia is prescribed for people with tuberculosis (TB), but is still not known which compound(s) acts against M. tuberculosis and its mode of action. The aim of this study was to identify and isolate antimycobacterial compounds from S. frutescens extracts against shikimate kinase, a drug target for M. tuberculosis. METHODS: S. frutescens were dried, ground and extracted with ethanol, dichloromethane: methanol and water. Fractionation and separation of compounds was done with column chromatography. Chromatograms were developed in butanol/acetic acid/water (BAW) [21:6:3]; chloroform/methanol/water/formic acid (CMWF1) [60:15:2:1] and (CMWF2) [21:9:1:0.3]. Separation and isolation of active compounds were done using preparative HPLC. The activity of the plant extracts were also screened against shikimate kinase enzyme (MtbSK) using the MtbSK inhibition assay. RESULTS: The DCM: MeOH (1:1) extract showed a high percentage inhibition (with an IC50 of 0.1 µg/ml) of MtbSK and the purified inhibitor was an Alpha-Linolenic Acid (ALA) compound and it had a significant IC50 of 3.7 µg/ml. CONCLUSIONS: This study demonstrated that ALA from S. frustescens is an inhibitor of shikimate kinase a good drug target for M. tuberculosis.

Antibacterial, antifungal and antioxidant activity of Olea africana against pathogenic yeast and nosocomial pathogens.

BACKGROUND: Olea africana leaves are used by Bapedi people to treat different ailments. The use of these leaves is not validated, therefore the aim of this study is to validate antimicrobial properties of this plant. METHODS: The ground leaves were extracted using solvents of varying polarity (hexane, chloroform, dichloromethane (DCM), ethyl acetate, acetone, ethanol, methanol, butanol and water). Thin layer chromatography (TLC) was used to analyse the chemical constituents of the extracts. The TLC plates were developed in three different solvent systems, namely, benzene/ethanol/ammonium solution (BEA), chloroform/ethyl acetate/formic acid (CEF) and ethyl acetate/methanol/water (EMW). The micro-dilution assay and bioautography method were used to evaluate the antibacterial activity of the extracts against Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis and Staphylococcus aureus and the antifungal activity against Candida albicans and Cryptococcus neoformans. RESULTS: Methanol was the best extractant, yielding a larger amount of plant material whereas hexane yielded the least amount. In phytochemical analyses, more compounds were observed in BEA, followed by EMW and CEF. Qualitative 2, 2- diphenylpacryl-1-hydrazyl (DPPH) assay displayed that all the extracts had antioxidant activity. Antioxidant compounds could not be separated using BEA solvent system while with CEF and EMW enabled antioxidant compounds separation. The minimum inhibitory concentrations (MIC) values against test bacteria ranged between 0.16 and 2.50 mg/mL whereas against fungi, MIC ranged from 0.16 to 0.63 mg/mL. Bioautography results demonstrated that more than one compound was responsible for antimicrobial activity in the microdilution assay as the compounds were located at different Rf values. CONCLUSIONS: The results indicate that leaf extracts of Olea africana contain compounds with antioxidant, antibacterial and antifungal activities. Therefore, further studies are required to isolate the active compounds and perform other tests such as cytotoxicity. Olea africana may be a potential source of antimicrobial compounds.

In Vitro Assessment of Cytotoxicity, Antioxidant, and Anti-Inflammatory Activities of Ricinus communis (Euphorbiaceae) Leaf Extracts.

Ricinus communis has been utilized traditionally as medicine to treat inflammatory related diseases including wounds, sores, and boils. The leaves of R. communis were sequentially extracted with n-hexane, dichloromethane, acetone, and methanol using serial exhaustive extraction method. Antioxidant activity of all crude extracts was quantitatively measured against 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) free radical molecules using ABTS(+) assay. Cytotoxic effect and anti-inflammatory activity of R. communis leaves extracts were evaluated on Human Caucasian skin fibroblast and Raw 264.7 macrophage cell lines, respectively. Methanol extract had the highest percentage free radical (ABTS(+)) scavenging activity of 95% at 2.50 mg/mL, acetone 91%, dichloromethane 62%, and hexane the least (50%). Percentage scavenging activity of ABTS(+) free radical molecules increases with increase in concentrations of the plant extracts. Hexane and dichloromethane extracts had more than 90% cell viability at 100 µg/mL after 24 and 48 hours of exposure. Methanol extract had LC50 of 784 µg/mL after 24-hour exposure, hexane had 629.3 µg/mL and dichloromethane 573.6 µg/mL, and 544.6 µg/mL was the lowest with acetone extract. The study present the first report on the scavenging activity of R. communis leaf extracts against ABTS(+) radicals and cytotoxic effects on human Caucasian skin fibroblast cell lines.

Anti-bacterial and anti-oxidant activities of leaf extracts of Combretum vendae (Combretecacea) and the isolation of an anti-bacterial compound.

BACKGROUND: Combretum vendae A.E. van Wyk (Combretaceae) is used for the treatment of bacterial related infections and oxidative related diseases by indigenous people of South Africa. Dried leaves extracts of C. vendae were investigated for bioactivity against a variety of bacterial strains and their antioxidant potential evaluated. MATERIALS AND METHODS: Constituents of leaf material were serially extracted using solvents of varying polarities, TLC chromatograms of the fractions were sprayed with 2,2 diphenyl-1-picrylhydrazyl (DPPH) to determine the presence of antioxidant compounds. Bio-autography was used to determine the number of antibacterial compounds active against Staphylococcus aureus, Enterococcus faecalis, Eschericha coli and Pseudomonas aeruginosa. Minimum inhibitory concentration (MIC) values were determined using serial microplate dilution method. The chloroform fraction was subjected to bio-assay guided column chromatography to isolate the active compound. RESULTS: The mass extracted by different solvents was below 10% dry weight. MIC values for different extracts against different pathogens ranges from 0.08 to 0.64 mg/ml. The compound isolated was identified as acacetin having an Rf value of 0.28 following elution in the Ethanol: Methanol: Water [E: M: W (10: 1.35: 1 v/v). Acacetin had MIC values ranging from 0.16 to 0.35 mg/ml. CONCLUSION: We report for the first time the isolation of acacetin as the main antibacterial compound from the leaves of Combretum vendae.

Validation of antimycobacterial plants used by traditional healers in three districts of the limpopo province (South Africa).

The aim of the study was to scientifically evaluate the antimycobacterial activity of selected indigenous medicinal plants from the Limpopo Province used for the treatment of humans with symptoms of Mycobacterium tuberculosis. The leaves of five plant species (Apodytes dimidiata, Artemisia, Combretum hereroense, Lippia javanica, and Zanthoxylum capense) were collected from the Lowveld National Botanical Garden in Nelspruit, South Africa. The dried leaves were powdered and extracted using hexane, dichloromethane, acetone, and methanol. Antimycobacterial activity was evaluated using microdilution assay and bioautography and ρ -iodonitrotetrazolium violet (INT) as indicator. Antioxidant activities were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH). Phytochemical content of extracts was further evaluated. The acetone extracts of L. javanica displayed antioxidant activity on BEA chromatogram. T Acetone extracts of A. afra had MIC value of 0.39 mg/mL against Mycobacterium smegmatis ATCC 1441. Acetone extracts of C. hereroense and L. javanica had MIC value of 0.47 mg/mL. Four bands that inhibited the growth of M. smegmatis were observed at R f values of 0.12, 0.63, and 0.87 on BEA and 0.73 on EMW. The plant species A. dimidiata, A. afra, C. hereroense, and L. javanica in this study demonstrated their potential as sources of anti-TB drug leads.