Ibuprofen as an adjuvant to conventional antimicrobials and essential oil compounds against skin pathogens.

AIMS: Antimicrobial resistance continues to be a growing concern, resulting in increased use of drug combinations. Antibiotic adjuvants are an emerging strategy that may potentiate an antibiotics efficacy. Ibuprofen's polypharmacological properties have been investigated for their antimicrobial and host-modulating potential. This study aimed to investigate the potential of a novel multidrug combination involving ibuprofen, essential oil compounds (EOCs), and conventional antimicrobials against skin pathogens. METHODS AND RESULTS: The minimum inhibitory concentrations of ibuprofen, conventional antimicrobials, and EOCs were determined and then combined and tested against 14 (reference and clinical) skin pathogens. The cytotoxicity was analysed using the MTT assay, whilst the anti-inflammatory effects were evaluated using lipopolysaccharide activated RAW264.7 murine macrophages. Four pairwise (Ibuprofen and antibiotic) (ΣFIC 0.33-0.50) and three triple (Ibuprofen and antibiotic with EOC) (ΣFIC 0.44-0.47) synergistic antimicrobial interactions were identified. These combinations demonstrated cell viability of 77.59%-100%. No combination significantly reduced nitric oxide production. CONCLUSION: The results from this study provide insight into the potential of a multidrug combination involving ibuprofen with conventional antimicrobials and EOCs against common skin pathogens.

The interactive effects of medicinal dyes with conventional antimicrobials against skin pathogens.

AIMS: This study aimed to explore potential synergistic effects of medicinal dyes with antimicrobials against pathogens responsible for skin infections. METHODS AND RESULTS: Antimicrobial testing was conducted using minimum inhibitory concentrations and minimum bactericidal/fungicidal concentration assays. The fractional inhibitory index (ΣFIC) of combinations was calculated, and isobolograms were constructed on selected combinations. Toxicity studies were conducted using the brine-shrimp lethality assay. Combination (1:1 ratio) studies noted that 26% of dye-antibiotic combinations were synergistic against the Gram-positive strains, 15% against the Gram-negative strains, and 14% against the yeasts. The Mercurochrome: Betadine® combination noted synergy at ratios against all the Staphylococcus aureus strains with ΣFIC values ranging from 0.05 to 0.48. The combination of Gentian violet with Gentamycin noted a 15-fold decrease in toxicity, and a selectivity index of 977.50 against the Escherichia coli (DSM 22314) strain. Time-kill studies were conducted on the combinations with the highest safe selectivity index (SI) value and lowest safe SI value i.e. Gentian violet with Gentamycin and Malachite green with Neomycin. Both combinations demonstrated better antimicrobial activity in comparison to the independent values and the controls. CONCLUSION: This study highlights the potential for medicinal dye combinations as a treatment for skin infections.

Copper(II)-photocatalyzed Hydrocarboxylation of Schiff bases with CO2: antimicrobial evaluation and in silico studies of Schiff bases and unnatural α-amino acids.

We synthesized and characterized two copper(II) complexes: [CuL2Cl]Cl and [CuL'2Cl]Cl, where L = 2,2'-bipyridine and L' = 4,4'-dimethyl-2,2'-bipyridine. We evaluated their photocatalytic hydrocarboxylation properties on a series of synthesized Schiff bases (SBs): (E)-1-(4-((5-bromo-2-hydroxybenzylidene)amino)phenyl)ethanone (SB1), (E)-N-(4-(dimethylamino)benzylidene)benzo[d]thiazol-2-amine (SB2), (E)-4-Bromo-2-((thiazol-2-ylimino)methyl)phenol (SB3), and (E)-4-((5-bromo-2-hydroxybenzylidene)amino)-1,5-dimethyl-2-phenyl-1H-pyrazol-3(2H)-one (SB4). Under mild photocatalytic reaction conditions (room temperature, 1 atm CO2, 30-watt Blue LED light), the derivatives of α-amino acids UAA1-4 were obtained with yields ranging from 5% to 44%. Experimental results demonstrated that [CuL2Cl]Cl exhibited superior photocatalytic efficiency compared to [CuL'2Cl]Cl, attributed to favourable electronic properties. In silico studies revealed strong binding strengths with E. faecalis DHFR (4M7U) for docked Schiff bases (SB) and unnatural α-amino acids (UAAs). In vitro studies further demonstrated significant antimicrobial and antifungal activity for SB2, SB3, and SB4, while none of the synthesized UAAs exhibited such properties, primarily due to the electronic and binding properties of these molecules.Communicated by Ramaswamy H. Sarma.

Optimizing the Antimicrobial Synergism of Melaleuca alternifolia (Tea Tree) Essential Oil Combinations for Application against Respiratory Related Pathogens.

Antimicrobial research into the use of Melaleuca alternifolia essential oil has demonstrated broad-spectrum activity; however, much of the research published focuses on identifying the potential of this essential oil individually, rather than in combination for an enhanced antimicrobial effect. This study aimed to determine the antimicrobial activity of four essential oil combinations, all inclusive of M. alternifolia, against nine pathogens associated with the respiratory tract. The minimum inhibitory concentration assay was used to determine the antimicrobial activity of four essential oil combinations, M. alternifolia in combination with Cupressus sempervirens, Origanum majorana, Myrtus communis, and Origanum vulgare essential oils. The interactions between essential oil combinations were analyzed using isobolograms and SynergyFinder 2.0 software to visualize the synergistic potential at varied ratios. The antimicrobial activity of the different combinations of essential oils all demonstrated the ability to produce an enhanced antimicrobial effect compared to the essential oils when investigated independently. The findings of this study determined that isobolograms provide a more in-depth analysis of an essential oil combination interaction; however, the value of that interaction should be further quantified using computational modelling such as SynergyFinder. This study further supports the need for more studies where varied ratios of essential oils are investigated for antimicrobial potential.

Bioautography-guided HPTLC-MS as a rapid hyphenated technique for the identification of antimicrobial compounds from selected South African Combretaceae species.

INTRODUCTION: Many species within Combretaceae are traditionally used for the treatment of bacterial infections. The similarity in chemistry and antimicrobial activities within the family pose a challenge in selecting suitable species for herbal drug development. OBJECTIVE: This study aimed at rapidly identifying antimicrobial compounds using bioautography-guided high-performance thin-layer chromatography coupled with mass spectrometry (HPTLC-MS). METHODS: Hierarchical cluster analysis of ultra-performance liquid chromatography-mass spectrometry data from the methanol extracts of 77 samples, representing four genera within Combretaceae, was carried out. Based on groupings on the dendrogram, 15 samples were selected for bioautography analysis against four pathogens (Staphylococcus aureus, Bacillus cereus, Escherichia coli and Salmonella typhimurium). Active compounds were identified using HPTLC-MS analysis of bands corresponding to the inhibition zones. RESULTS: Bioautography revealed 15 inhibition zones against the four pathogens, with the most prominent present for Combretum imberbe. Analysis of the active bands, using HPTLC-MS indicated that flavonoids, triterpenoids and combretastatin B5 contributed to the antibacterial activity. The compounds corresponding to molecular ions m/z 471 (Combretum imberbe) and 499 (Combretum elaeagnoides) inhibited all four pathogens, and were identified as imberbic acid and jessic acid, respectively. Chemotaxonomic analysis indicated that arjunic acid, ursolic acid and an unidentified triterpenoid (m/z 471) were ubiquitous in the Combretaceae species and could be responsible for their antibacterial activities. CONCLUSION: Application of HPTLC-MS enabled the rapid screening of extracts to identify active compounds within taxonomically related species. This approach allows for greater efficiency in the natural product research workflow to identify bioactive compounds in crude extracts.

Antibacterial Screening, Biochemometric and Bioautographic Evaluation of the Non-Volatile Bioactive Components of Three Indigenous South African Salvia Species.

Salvia africana-lutea L., S. lanceolata L., and S. chamelaeagnea L. are used in South Africa as traditional medicines to treat infections. This paper describes an in-depth investigation into their antibacterial activities to identify bioactive compounds. Methanol extracts from 81 samples were screened against seven bacterial pathogens, using the microdilution assay. Biochemometric models were constructed using data derived from minimum inhibitory concentration (MIC) and ultra-performance liquid chromatography-mass spectrometry data. Active molecules in selected extracts were tentatively identified using high-performance thin layer chromatography (HPTLC), combined with bioautography, and finally, by analysis of active zone eluates by mass spectrometry (MS) via a dedicated interface. Salvia chamelaeagnea displayed notable activity towards all seven pathogens, and the activity, reflected by MICs, was superior to that of the other two species, as confirmed through ANOVA. Biochemometric models highlighted potentially bioactive compounds, including rosmanol methyl ether, epiisorosmanol methyl ether and carnosic acid. Bioautography assays revealed inhibition zones against A. baumannii, an increasingly multidrug-resistant pathogen. Mass spectral data of the eluted zones correlated to those revealed through biochemometric analysis. The study demonstrates the application of a biochemometric approach, bioautography, and direct MS analysis as useful tools for the rapid identification of bioactive constituents in plant extracts.

Investigating the Antituberculosis Activity of Selected Commercial Essential Oils and Identification of Active Constituents Using a Biochemometrics Approach and In Silico Modeling.

Tuberculosis (TB) is a disease caused by Mycobacterium tuberculosis which has become prevalent due to the emergence of resistant M. tuberculosis strains. The use of essential oils (EOs) as potential anti-infective agents to treat microbial infections, including TB, offers promise due to their long historical use and low adverse effects. The current study aimed to investigate the in vitro anti-TB activity of 85 commercial EOs, and identify compounds responsible for the activity, using a biochemometrics approach. A microdilution assay was used to determine the antimycobacterial activity of the EOs towards some non-pathogenic Mycobacterium strains. In parallel, an Alamar blue assay was used to investigate antimycobacterial activity towards the pathogenic M. tuberculosis strain. Chemical profiling of the EOs was performed using gas chromatography-mass spectrometry (GC-MS) analysis. Biochemometrics filtered out putative biomarkers using orthogonal projections to latent structures discriminant analysis (OPLS-DA). In silico modeling was performed to identify potential therapeutic targets of the active biomarkers. Broad-spectrum antimycobacterial activity was observed for Cinnamomum zeylanicum (bark) (MICs = 1.00, 0.50, 0.25 and 0.008 mg/mL) and Levisticum officinale (MICs = 0.50, 0.5, 0.5 and 0.004 mg/mL) towards M. smegmatis, M. fortuitum, M. gordonae and M. tuberculosis, respectively. Biochemometrics predicted cinnamaldehyde, thymol and eugenol as putative biomarkers. Molecular docking demonstrated that cinnamaldehyde could serve as a scaffold for developing a novel class of antimicrobial compounds by targeting FtsZ and PknB from M. tuberculosis.

Optimization of Antioxidant Synergy in a Polyherbal Combination by Experimental Design.

Culinary herbs and spices are known to be good sources of natural antioxidants. Although the antioxidant effects of individual culinary herbs and spices are widely reported, little is known about their effects when used in combination. The current study was therefore undertaken to compare the antioxidant effects of crude extracts and essential oils of some common culinary herbs and spices in various combinations. The antioxidant interactions of 1:1 combinations of the most active individual extracts and essential oils were investigated as well as the optimization of various ratios using the design of experiments (DoE) approach. The 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays were used to determine the antioxidant activity, and MODDE 9.1® software (Umetrics AB, Umea, Sweden) was used to determine the DoE. The results revealed synergism for the following combinations: Mentha piperita with Thymus vulgaris methanol extract (ΣFIC = 0.32 and ΣFIC = 0.15 using the DPPH and FRAP assays, respectively); Rosmarinus officinalis with Syzygium aromaticum methanol extract (ΣFIC = 0.47 using the FRAP assay); T. vulgaris with Zingiber officinalis methanol extracts (ΣFIC = 0.19 using the ABTS assay); and R. officinalis with Z. officinalis dichloromethane extract (ΣFIC = 0.22 using the ABTS assay). The DoE produced a statistically significant (R2 = 0.905 and Q2 = 0.710) model that was able to predict extract combinations with high antioxidant activities, as validated experimentally. The antioxidant activities of the crude extracts from a selection of culinary herbs and spices were improved when in combination, hence creating an innovative opportunity for the future development of supplements for optimum health.

Antimicrobial Formulation of Chrysopogon zizanioides Essential Oil in an Emulsified Lotion for Acne.

Acne is a skin condition arising from excess sebum production and microbial overgrowth within the pilosebaceous unit. Several commercial essential oils have shown promising activity against acne-related pathogens. Due to their volatility and thermal instability, the formulation of essential oils into commercial products remains a pharmaceutical challenge. Thus, this study aimed to develop a viable anti-acne topical treatment as an oil-in-water emulsified lotion to overcome these challenges. Chrysopogon zizanioides (vetiver) displayed noteworthy antimicrobial activity with a mean minimum inhibitory concentration of 0.14 mg/mL against Cutibacterium acnes, Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus pyogenes. Emulsified lotions containing C. zizanioides were developed through the hydrophilic-lipophilic balance approach. At tested hydrophilic-lipophilic balance values of 8, 9, and 10, C. zizanioides emulsified lotions displayed maximum stability at hydrophilic-lipophilic balance 9 with a minimum change in mean droplet size and polydispersity index of 20.61 and 33.33%, respectively, over 84 days. The C. zizanioides emulsified lotion at optimum hydrophilic-lipophilic balance 9 completely inhibited the growth of C. acnes and killed S. aureus, S. epidermidis, and S. pyogenes within 24 h. Additionally, the lotion retained antimicrobial activity against these test micro-organisms over the 84-day stability test period. Thus, the C. zizanioides emulsified lotion demonstrated physical stability and antimicrobial efficiency, making it an ideal natural product anti-acne treatment.

The Antimicrobial and Toxicity Influence of Six Carrier Oils on Essential Oil Compounds.

Essential oil compounds have been identified as alternative antimicrobials; however, their use is limited due to their toxicity on human lymphocytes, skin, and reproduction. Carrier oils can reduce the toxicity of essential oils, which raises the question as to whether such activity would extend to the essential oil compounds. Thus, this study aimed to investigate the antimicrobial and toxicity activity of essential oil compounds in combination with carrier oils. The antimicrobial properties of the essential oil compounds, alone and in combination with carrier oils, were determined using the broth microdilution assay. The toxicity was determined using the brine shrimp lethality assay. Antimicrobial synergy (ΣFIC ≤ 0.50) occurred in 3% of the samples when tested against the ESKAPE pathogens. The compound thymoquinone in combination with the carrier oil Prunus armeniaca demonstrated broad-spectrum synergistic activity and a selectivity index above four, highlighting this combination as the most favorable. The carrier oils reduced the toxicity of several compounds, with Calendula officinalis and P. armeniaca carrier oils being responsible for the majority of the reduced toxicity observed. This study provides insight into the interactions that may occur when adding a carrier oil to essential oil compounds.

Essential Oil Blends: The Potential of Combined Use for Respiratory Tract Infections.

This study investigated the potential efficacy of 369 commercial essential oil combinations for antimicrobial, anti-toxic and anti-inflammatory activity with the aim of identifying synergy among essential oils commonly used in combination by aromatherapists for respiratory purposes. Essential oil combinations were assessed for their antimicrobial activities using a panel of Gram-positive, Gram-negative, and yeast strains associated with respiratory tract infections. The antimicrobial activity was measured by determining the minimal inhibitory concentration (MIC) of microbial growth. The fractional inhibitory concentration index (ΣFIC) was calculated to determine the antimicrobial interactions between the essential oils in the combination. The toxicity of the essential oil combinations was tested in vitro using the brine shrimp lethality assay, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on RAW 264.7 mouse macrophage cells and A549 lung cancer cell lines. In addition, an inflammatory response was evaluated measuring nitric oxide production. The essential oils, when in combination, demonstrated an increased antimicrobial effect, a reduction in toxicity and provided improved anti-inflammatory outcomes. Five distinct combinations [Cupressus sempervirens (cypress) in combination with Melaleuca alternifolia (tea tree), Hyssopus officinalis (hyssop) in combination with Rosmarinus officinalis (rosemary), Origanum marjorana (marjoram) in combination with M. alternifolia, Myrtus communis (myrtle) in combination with M. alternifolia and Origanum vulgare (origanum) in combination with M. alternifolia] were found to be the most promising, demonstrating antimicrobial activity, reduced cytotoxicity and improved anti-inflammatory effects. With the increased prevalence of respiratory tract infections and the growing antimicrobial resistance development associated with antimicrobial treatments, this study provides a promising complementary alternative for the appropriate use of a selection of essential oil combinations for use in the respiratory tract.

Three-Dimensional Printability of an ECM-Based Gelatin Methacryloyl (GelMA) Biomaterial for Potential Neuroregeneration.

The current study introduces two novel, smart polymer three-dimensional (3D)-printable interpenetrating polymer network (IPN) hydrogel biomaterials with favorable chemical, mechanical, and morphological properties for potential applications in traumatic brain injury (TBI) such as potentially assisting in the restoration of neurological function through closure of the wound deficit and neural tissue regeneration. Additionally, removal of injury matter to allow for the appropriate scaffold grafting may assist in providing a TBI treatment. Furthermore, due to the 3D printability of the IPN biomaterials, complex structures can be designed and fabricated to mimic the native shape and structure of the injury sight, which can potentially assist with neural tissue regeneration after TBI. In this study, a peptide-only approach was employed, wherein collagen and elastin in a blend with gelatin methacryloyl were prepared and crosslinked using either Irgacure or Irgacure and Genipin to form either a semi or full IPN hydrogel 3D-printable neuromimicking platform system, respectively. The scaffolds displayed favorable thermal stability and were amorphous in nature with high full width at half-maximum values. Furthermore, no alteration to the peptide secondary structure was noted using Fourier transform infrared spectroscopy. The IPN biomaterials have a stiffness of around 600 Pa and are suitable for softer tissue engineering applications-that is, the brain. Scanning electron micrographs indicated that the IPN biomaterials had a morphological structure with a significant resemblance to the native rat cortex. Both biomaterial scaffolds were shown to support the growth of PC12 cells over a 72 h period. Furthermore, the increased nuclear eccentricity and nuclear area were shown to support the postulation that the IPN biomaterials maintain the cells in a healthy state encouraging cellular mitosis and proliferation. The Genipin component of the full IPN was further shown to exhibit antimicrobial properties and this suggests that Genipin can prevent the growth of pathogens associated with postsurgical brain infections. In addition to these findings, the study presents an anomaly, wherein the full IPN is found to be more brittle than the semi IPN, a finding that is in contradiction with the literature. This research, therefore, contributes to the collection of potential biomaterials for TBI applications coupled with 3D printing and can assist in the progression of neural treatments toward patient-specific scaffolds through the development of custom scaffolds.

The use of chemometric modelling to determine chemical composition-antimicrobial activity relationships of essential oils used in respiratory tract infections.

The antimicrobial effects of essential oils are commonly cited within aromatherapeutic texts for use in respiratory tract infections. These essential oils are inhaled or applied to the skin to treat infections and manage symptoms associated with these conditions. A limited number of these essential oils have been scientifically studied to support these claims, specifically, against respiratory pathogens. This study reports on the minimum inhibitory concentration (MIC) of 49 commercial essential oils recommended for respiratory tract infections, and identifies putative biomarkers responsible for the determined antimicrobial effect following a biochemometric workflow. Essential oils were investigated against nine pathogens. Three essential oils, Amyris balsamifera (amyris), Coriandrum sativum (coriander) and Santalum austrocaledonicum (sandalwood) were identified as having greater activity (MIC value = 0.03-0.13 mg/ml) compared to the other essential oils investigated. The essential oil composition of all 49 oils were determined using Gas Chromatography coupled to Mass Spectroscopy (GC-MS) analysis and the GC-MS data analysed together with the antimicrobial data using chemometric tools. Eugenol was identified as the main biomarker responsible for antimicrobial activity in the majority of the essential oils. The ability of a chemometric model to accurately predict the active and inactive biomarkers of the investigated essential oils against pathogens of the respiratory tract was 80.33%.

A Review of Plant-Based Therapies for the Treatment of Urinary Tract Infections in Traditional Southern African Medicine.

Urinary tract infections (UTIs) are amongst the most common bacterial infections globally, with ∼11% of the world's population contracting at least one infection annually. Several South African plants are used in traditional healing systems to treat UTIs, yet the therapeutic potential of these plants against bacteria that cause UTI remains poorly explored. This study documents southern African plant species used traditionally to treat UTIs. An extensive literature review was undertaken to document the southern African plant species that are used in traditional South African medicine to treat UTIs, thereby highlighting gaps in the current research that require further study. One hundred and fifty-three southern African plant species that are used to treat UTIs were identified. Eighty-five southern African plants were identified as having noteworthy inhibitory activity against the major UTI-causing bacteria. Few of those studies screened against all of the bacterial causes of UTIs, and none of those studies examined the mechanism of action of the plant preparations. Furthermore, many of those studies did not test the toxicity of the plant extracts, so an evaluation of the safety for therapeutic usage was lacking. Substantial further research is to determine their potential for therapeutic use.

Volatile phenolics: A comprehensive review of the anti-infective properties of an important class of essential oil constituents.

Historically, essential oils and their lead molecules have been extensively recognised for their anti-infective properties. In this context, certain volatile phenolics (VPs) have emerged as important antimicrobial compounds with excellent inhibitory activity against pathogenic bacteria and fungi, which further extends to drug-resistant and biofilm-forming micro-organisms. In this review, we aim to collate and discuss a number of published papers on the anti-infective activities of naturally occurring VPs with special emphasis on eugenol, isoeugenol, thymol and carvacrol, using Scopus Web of Science and PubMed databases. The biosynthesis and extraction of these VPs are discussed, while particular attention is given to their broad-spectrum antimicrobial activity and the mechanisms of action. We highlight combinational studies of the VPs with other phytocompounds and with commercially available drugs, which may be a promising and a rewarding future approach to combat antimicrobial resistance. These VPs alone, or concomitantly with other compounds or drugs, have the potential to be incorporated into different formulations for biomedical applications. An in-depth assessment of 2310 articles retrieved from the Scopus database spanning a 35-year period indicated 23.1% increase in global publication growth in VPs anti-infective research, with authors from Italy, Portugal and Austria dominating the research landscape. The dominant areas of investigations are identified as antimicrobial activity, antibacterial mechanism of action, antifungal mechanism of action, extraction methods and phytochemistry, use in the food industry, and for oral and dental anti-infective activity. Specific research areas, which require future attention include; antituberculosis research, nanoparticle formulation of antimicrobial active VP molecules, preclinical and clinical trials. The antimicrobial testing of isoeugenol was found to be the least studied of the VPs and this requires further attention.

South Africa's Best BARK Medicines Prescribed at the Johannesburg Muthi Markets for Skin, Gut, and Lung Infections: MIC's and Brine Shrimp Lethality.

Indigenous trade of medicinal plants in South Africa is a multi-million-rand industry and is still highly relevant in terms of primary health care. The purpose of this study was to identify today's most traded medicinal barks, traditionally and contemporaneously used for dermatological, gastrointestinal, and respiratory tract infections; then, to investigate the antimicrobial activity and toxicity of the respective extracts and interpret outcomes in light of pharmacokinetics. Thirty-one popularly traded medicinal barks were purchased from the Faraday and Kwa Mai-Mai markets in Johannesburg, South Africa. Information on the medicinal uses of bark-based medicines in modern commerce was recorded from randomly selected traders. The minimum inhibitory concentration (MIC) method was used for antimicrobial screening, and brine shrimp lethality was used to determine toxicity. New medicinal uses were recorded for 14 bark species. Plants demonstrating some broad-spectrum activities against tested bacteria include Elaeodendron transvaalense, Erythrina lysistemon, Garcinia livingstonei, Pterocelastrus rostratus, Rapanea melanophloeos, Schotia brachypetala, Sclerocarya birrea, and Ziziphus mucronata. The lowest MIC value of 0.004 mg/mL was observed against Staphylococcus epidermidis for a dichloromethane bark extract of E. lysistemon. The tested medicinal barks were shown to be non-toxic against the Artemia nauplii (brine shrimp) bioassay, except for a methanol extract from Trichilia emetica (69.52% mortality). Bacterial inhibition of bark extracts with minimal associated toxicity is consistent with the safety and valuable use of medicinal barks for local muthi market customers. Antimicrobial outcomes against skin and gastrointestinal pathogens are feasible because mere contact-inhibition is required in vivo; however, MIC values against respiratory pathogens require further explaining from a pharmacokinetics or pharmacodynamics perspective, particularly for ingested rather than smoked therapies.

Investigating antimicrobial compounds in South African Combretaceae species using a biochemometric approach.

ETHNOPHARMACOLOGICAL RELEVANCE: Many species within the family Combretaceae are popular medicinal plants that are used traditionally to treat various conditions, of which many are related to bacterial infections. Global concerns regarding the increasing resistance of pathogens towards currently available antibiotics have encouraged researchers to find new drugs with antibacterial activity, particularly from plant sources. AIM OF THE STUDY: This study was aimed at exploring the broad-spectrum antibacterial potential of methanol extracts of species representing four genera of Combretaceae (Combretum, Pteleopsis, Quisqualis, Terminalia), indigenous to South Africa, using a biochemometric approach. MATERIALS AND METHODS: The microdilution assay was used to determine the antibacterial activities, measured as minimum inhibitory concentrations (MICs), of the 51 methanol extracts representing 35 Combretaceae species, against nine species of pathogenic bacteria. Integrative biochemometric analysis was performed, thereby correlating the MIC values with the metabolomic data obtained from ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) analysis. Orthogonal projections to latent structures-discriminant analysis (OPLS-DA) models were constructed for six pathogens displaying variation in their susceptibility towards the extracts. RESULTS: Evaluation of the overall MIC values obtained indicated that extracts of species from the four genera displayed the highest activity towards Bacillus cereus ATCC 11778 (average MIC 0.52 mg/mL) and Salmonella typhimurium ATCC 14028 (average MIC 0.63 mg/mL). These bacteria were the most sensitive Gram-positive and Gram-negative bacteria, respectively. Extracts from Combretum acutifolium, Combretum imberbe and Combretum elaeagnoides were the most active, with average MIC values of 0.70 mg/mL, 0.52 mg/mL and 0.45 mg/mL, respectively. Five triterpenoid compounds were tentatively identified as biomarkers from the biochemometric analysis. CONCLUSION: Correlation of the phytochemistry of species from four genera in the Combretaceae family with antibacterial activity revealed that triterpenoids are responsible for the broad-spectrum antibacterial activity observed.

Essential Oil Variation within Warburgia salutaris-A Coveted Ethnomedicinal Aromatic Tree.

Warburgia salutaris, known as 'Pepper bark', is an ethnomedicinally important tree found in the southern regions of Africa. A total of 75 fresh leaf specimens of W. salutaris (n=40 wild and 35 cultivated) were collected from the Limpopo (wild) and KwaZulu-Natal provinces (cultivated), two distinct locations in South Africa. In this study, the leaf essential oils obtained by hydrodistillation were characterized using gas chromatography coupled to mass spectrometry/flame ionization detection (GC/MS/FID). More than 15 compounds, accounting for 90-99 % of the total oil composition were identified. The analysis revealed that myrcene (0.6-65.3 %), (E)-ß-ocimene (nd-56.9 %), (Z)-ß-ocimene (nd-19.1 %), α-pinene (nd-19.1 %) and limonene (nd-11.7 %) are major constituents of W. salutaris essential oils. Chemometric analysis revealed two major chemotypes within the essential oils with a modeled variation of approximately 60 %. Linalool and germacrene D were revealed as markers associated with the wild-harvested oils, while cultivated oils were distinguished by higher levels of limonene and α-humulene. The intra-population variation indicated two chemically distinct chemotypes from three different populations, however, the season of harvest did not have a direct influence on the chemical profiles of the essential oils.

Unravelling the Antibacterial Activity of Terminalia sericea Root Bark through a Metabolomic Approach.

Terminalia sericea Burch. ex. DC. (Combretaceae) is a popular remedy for the treatment of infectious diseases. It is widely prescribed by traditional healers and sold at informal markets and may be a good candidate for commercialisation. For this to be realised, a thorough phytochemical and bioactivity profile is required to identify constituents that may be associated with the antibacterial activity and hence the quality of raw materials and consumer products. The aim of this study was to explore the phytochemistry and identify the antibacterial constituents of T. sericea root bark, using a metabolomic approach. The chemical profiles and antibacterial activities of 42 root bark samples collected from three districts in the Limpopo Province, South Africa, were evaluated. Dichloromethane:methanol (1:1) extracts were analysed using ultraperformance liquid chromatography (UPLC)-mass spectrometry (MS), and chemometric models were constructed from the aligned data. The extracts were tested against Bacillus cereus (ATCC 11778), Staphylococcus epidermidis (ATCC 12223), Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 8739), Klebsiella pneumoniae (ATCC 13883), Pseudomonas aeruginosa (ATCC 27853), Shigella sonnei (ATCC 9292) and Salmonella typhimurium (ATCC 14028), using the minimum inhibition microdilution assay. Nine compounds; sericic acid, sericoside, resveratrol-3-O-ß-rutinoside, ellagic acid, flavogallonic acid dilactone, methyl-flavogallonate, quercetin-3-(2''-galloylrhamnoside), resveratrol-3-(6''-galloyl)-O-ß-d-glucopyranoside and arjunetin, were isolated from the root bark. All the compounds, with the exception of sericic acid, sericoside and resveratrol-3-O-ß-rutinoside, were isolated for the first time from the root bark of T. sericea. Chemometric analysis revealed clustering that was not population specific, and the presence of three groupings within the samples, characterised by sericic acid, sericoside and an unidentified compound (m/z 682/4.66 min), respectively. The crude extracts from different populations displayed varied antibacterial activities against S. typhimurium (minimum inhibitory concentrations (MICs) 0.25-1.0 mg/mL), but similar activity towards Bacillus cereus (1.0 mg/mL). Several compounds present in the root bark were highly active towards all or most of the pathogens tested, but this activity was not reflected by the chemical profiles of extracts prepared from the individual samples. Among the pure compounds tested, only flavogallonic acid dilactone and methyl-flavogallonate exhibited broad-spectrum activity. A biochemometric analysis indicated that there was no consistent association between the levels of phytochemicals and the activity of the active or non-active extracts. Although it was deduced that the major constituents of T. sericea root bark contributed to the chemotypic variation, further investigation of the interactions of compounds present in the root bark may provide antibacterial efficacies not evident when examining compounds singularly. The data reported herein will provide information that is fundamentally important for the development of quality control protocols.

The traditional use of southern African medicinal plants in the treatment of viral respiratory diseases: A review of the ethnobotany and scientific evaluations.

ETHNOPHARMACOLOGICAL RELEVANCE: Viral respiratory infections are amongst the most common infections globally, with most of the world's population contracting at least one infection annually. Numerous plant species are used in traditional southern African healing systems to treat these diseases and to alleviate the symptoms. Despite this, the therapeutic potential of these plants against viral respiratory diseases remains poorly explored. AIM OF THE STUDY: The aim of this study was to document the southern African plant species used in traditional medicine to treat viral respiratory infections. We also examined the extent of scientific evaluations of southern African plant species against the respiratory-infective viruses, with the aim of stimulating interest in this area and focusing on future studies. MATERIALS AND METHODS: We undertook an extensive review of ethnobotanical books, reviews and primary scientific studies to identify southern African plants which are used in traditional southern African medicine to treat viral respiratory diseases. This information was used to identify gaps in the current research that require further study. RESULTS: Two hundred and fifty-seven southern African plant species were identified as traditional therapies for viral respiratory diseases. Surprisingly, only one of those species (as well as twenty-one other species not recorded for these purposes) has been evaluated for the ability to block respiratory virus production. Furthermore, most of these studies screened against a single viral strain and none of those studies examined the mechanism of action of the plant preparations. CONCLUSIONS: Despite well documented records of the use of southern African plants to treat respiratory viral diseases, the field is poorly explored. Nearly all of the plant species used in traditional healing systems to treat these diseases are yet to be tested. Substantial further work is required to verify the efficacy of these traditional medicines.