The antimicrobial susceptibility patterns of diabetic foot ulcers in the South African public healthcare sector.

BACKGROUND: Diabetic foot syndrome is defined as the presence of a diabetic foot ulcer (DFU) associated with neuropathy, peripheral artery disease and infection. While the use of antimicrobials in the treatment of DFU infection remains a mainstay, the choice of antimicrobial remains problematic owing to the presence of multidrug-resistant polymicrobial infections. In the South African public healthcare sector, the treatment of DFUs is based on the Standard Treatment Guidelines (STGs) and the Essential Drug List. These guidelines are developed using evidence-based medicine and are based on global susceptibility patterns rather than local susceptibility data, and may not provide the most appropriate treatment options. OBJECTIVES: To determine the antimicrobial susceptibility patterns of DFUs isolated from patients visiting selected Gauteng provincial public hospitals in order to determine a clinically effective treatment protocol for the management of these infections. METHODS: Sample swabs were taken from 51 DFUs using the Levine method. Each sample swab was spread onto blood agar plates, and thereafter individual pathogens were isolated. The antimicrobial susceptibility patterns of all isolated pathogens were determined using zone of inhibition measurements. Pathogens were grouped according to macromorphological characteristics as well as susceptibility patterns, and a representative isolate from each group was then identified. RESULTS: A total of 51 DFU ulcer swabs from 45 patients were included in the study. From the sample swabs, a total of 445 pathogens were isolated. The most effective antimicrobial was found to be gentamicin, followed by ciprofloxacin. Amoxicillin/clavulanic acid, the first-line treatment according to the STGs, was found to be ineffective for many of the isolated pathogens. The most commonly isolated pathogens were Proteus mirabilis, Enterococcus faecalis and Pseudomonas aeruginosa. CONCLUSION: These findings demonstrate the urgent need to reassess the STGs and base treatment plans on local epidemiological data. This study provides valuable data on common causative pathogens in DFU infections, as well as the resistance patterns of these pathogens, forming a baseline on which to base future DFU treatment plans.

Antimicrobial activity of Southern African medicinal plants on Helicobacter pylori and Lactobacillus species.

ETHNOPHARMACOLOGICAL RELEVANCE: Numerous medicinal plants have been used traditionally in South Africa for gastric ulcer treatment. Helicobacter pylori is known for causing inflammation and the onset of gastric ulcers. While several studies explored medicinal plants against H. pylori, investigation of medicinal plants used for gastric ulcers has been neglected, as well as the effects these plants would have on bacteria occurring naturally in the gut microbiome. AIM OF THE STUDY: This study aimed to investigate Southern African medicinal plants used traditionally for treating gastric ulcers against H. pylori , as well as the effects that these plants have when combined with Lactobacillus species and tested against H. pylori. METHODOLOGY: Based on evidence from the ethnobotanical literature, 21 plants were collected. Their antimicrobial activity was assessed against five clinical H. pylori strains, and in combination with each of three Lactobacillus species, using the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) broth microdilution assays. Toxicity was assessed using the brine shrimp lethality assay. RESULTS: Noteworthy activity was observed against at least one H. pylori strain for 12 plant species. The lowest mean MICs were from organic extracts of Carissa edulis Vahl (0.18 mg/mL) and Chironia baccifera L. (0.20 mg/mL), and aqueous extracts of Sansevieria hyacinthoides (L.) Druce (0.26 mg/mL) and Dodonaea viscosa Jacq. (0.30 mg/mL). Aqueous extracts of the investigated plants were combined with Lactobacillus species, and the majority of combinations showed increased antimicrobial activity compared with the extracts alone. Combinations of Lactobacillus rhamnosus with 18 of the 21 aqueous plant extracts showed at least a two-fold decrease in the mean MBC against all H. pylori strains tested. Lactobacillus acidophilus combined with either Protea repens L., Carpobrotus edulis (L.) L. Bolus or Warburgia salutaris (Bertol.f.) Chiov. aqueous extracts had the best anti-H. pylori activity (mean MBCs of 0.10 mg/mL for each combination). Only four organic and one aqueous extract(s) were considered toxic. CONCLUSION: These results highlight the potential of medicinal plants to inhibit H. pylori growth and their role in traditional treatments for the management of ulcers. The results also indicate that aqueous extracts of these plants do not hinder the growth of bacteria that occur naturally in the gut microbiome and play a role in maintaining gut health, as well as show the potential benefit of including Lactobacillus species as potentiators of H. pylori activity.

Blood purification practices: Some ethnopharmacological insight from a rural community in KwaZulu-Natal, South Africa.

ETHNOPHARMACOLOGICAL RELEVANCE: Blood purification practices, also referred to as blood cleansing or detoxification, is an ancient concept which is widespread within African traditional medicine. There is, however, limited consolidated information about the selection of plant species, the purpose of use and how the preparation is made. An in-depth examination of how regional communities use medicinal plants for blood purification is needed to better understand this ethnopharmacological practice. AIM OF THE STUDY: To gather knowledge on the traditional use of medicinal plants used for the purpose of blood purification by laypersons in northern Maputaland, KwaZulu-Natal, South Africa. METHODS: An ethnobotanical survey was conducted on medicinal plants used in the rural areas of northern Maputaland for blood purification. Interviews were conducted in 55 homesteads, where the most knowledgeable participant per household was interviewed using structured questionnaires. Information collected included knowledge of the concept of blood purification, purpose for medicating, plant selection, and mode of preparation. The most important species and families were identified through indices. RESULTS: Sixty-four plant species from 35 families were recorded as blood purifiers for the treatment of ailments linked to the need for blood purification. Terminalia sericea Burch. ex DC. was the most used plant species (mentioned by 39% of the people interviewed). The conditions that were mostly associated with blood purification were skin complaints, gastro-intestinal disorders, and overall improvement of general health. Blood purification is also commonly used for non-medical purposes such as granting good luck or as a love charm. Six new plant species and five new Zulu vernacular names had not previously been documented for blood purification. CONCLUSION: The frequently mentioned use of medicinal plants for blood purification by the rural community of northern Maputaland clearly emphasises the importance of this cultural practice. There is no doubt that when examining African traditional medicine, holistic terms such as "blood cleansing" cannot be ignored. Future studies may provide a better understanding of this often misunderstood and undocumented practice.

The traditional use of Southern African medicinal plants to alleviate fever and their antipyretic activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Multiple plant species are used traditionally in southern Africa to treat fever. This review summarises the traditional use, highlights promising plant species and focuses on validation studies to highlight future areas of research. AIM OF THE STUDY: This study aimed to critically review the use of southern African plants to alleviate fever. Additionally, an examination of research into those plants was undertaken to highlight gaps in the literature with the aim of stimulating further research in this field. MATERIALS AND METHODS: Ethnobotanical books, reviews and primary research studies were searched to identify plant species traditionally used to treat fever in southern Africa and to summarise the current knowledge and identify areas requiring further research. This study was non-biased and did not have any taxonomic preferences, nor did it favour any southern African healing systems over the others. RESULTS: The traditional therapeutic uses of 203 southern African plant species to alleviate fever were recorded. Leaves and roots/bulbs/rhizomes were the most commonly used plant part, and they were most commonly prepared as decoctions or infusions and consumed orally. Notably, only twenty-two species (∼11% of the identified species) have been scientifically evaluated for antipyretic properties, with substantially fewer mechanistic studies evident. CONCLUSIONS: Despite the availability of ethnobotanical records, the antipyretic properties of southern African medicinal plants are poorly reported. Indeed, the efficacy of most plants is yet to be verified and very few mechanistic studies are available.

A scoping review of African natural products against gastric ulcers and Helicobacter pylori.

ETHNOPHARMACOLOGICAL RELEVANCE: African traditional medicine is one of the oldest and most diverse practices for treating ailments and numerous natural products have been recommended for gastric ulcer treatment. Helicobacter pylori is the main causative organism implicated in several diseases, most notably in causing inflammation and the onset of gastric ulcers. Current H. pylori treatment methods are losing efficacy as H. pylori rapidly gains resistance to antibiotics. Hence, a search into natural products and their historical traditional efficacy for the treatment of gastric ulcers is of interest. AIM OF THE STUDY: This review aimed to summarise the African use of natural products, including medicinal plants noted in ethnobotanical reviews, used traditionally to treat gastric ulcers, and highlights the investigations into the anti-H. pylori activity of medicinal plants and bee products found in Africa. METHODOLOGY: A systematic review was carried out to identify natural products, including those used traditionally in Africa to treat gastric ulcers, and to correlate this with scientific investigations into the anti-H. pylori activity of natural products used in Africa. RESULTS: A total of 107 literature sources describing the traditional use of medicinal plants in gastric ulcer treatment were found, from which 360 medicinal plants were identified. Of the plants used traditionally for gastric ulcer treatment, 11% were investigated either in vitro or in vivo for anti-ulcer and anti-H. pylori activity. Of the 122 medicinal plants eliciting antimicrobial or anti-ulcer activity, Hibiscus sabdariffa L. calyx extract and Terminalia macroptera Guill. & Perr. root extract were found to have the most noteworthy antimicrobial activity, with minimum inhibitory concentrations (MICs) of 0.01 mg/mL and 0.03 mg/mL respectively. The essential oils of Piper longum L. and Pachira aquatica Aubl. displayed the most notable in vitro anti-H. pylori activity (MIC of 0.01 mg/mL and 0.02 mg/mL). Several in vivo studies found medicinal plant extracts effective in reducing the H. pylori load along the gastric mucosa. The South African honey variants, Pure Honey and Champagne Royal Train (common names given by supplier) were the most antimicrobially effective (MIC of 0.01-10.0%, 0.63-10.00% v/v) in inhibiting H. pylori when assessed in vitro. CONCLUSION: These results highlight the potential of natural products to inhibit H. pylori growth and serve as a possible stepping-stone in understanding the management of ulcers. Furthermore, effective natural product treatment or prophylactic use for preventing H. pylori growth may provide a more affordable option for African populations.

Traditionally used polyherbals in a southern African therapeutic context.

ETHNOPHARMACOLOGICAL RELEVANCE: In African traditional medicine, there are several plant species that are used in combination with either other plant species or non-plant derived combinations such as sugar and honey, salt and vinegar, milk, fat etc. This review examines the role of these combinations and postulates the scientific and therapeutic validation of such combinations. AIM OF THE STUDY: This study reviewed the ethnopharmacological literature and documented the use of southern African plant combinations to find a scientific rationale for such combinations, and provide recommendations for future studies. MATERIALS AND METHODS: Ethnobotanical books and online databases such as Scopus, ScienceDirect, PubMed and Google Scholar were used to find ethnobotanical studies within the southern African context that focus on the combinations of plants with other plants or various additional ingredients. The scientific literature was examined to determine if evidence was available to substantiate such combinations. RESULTS: One hundred and eighty-seven medicinal plant (plant-to-plant) combinations that are used in the southern African traditional healing system were recorded. These plant combinations were used against infections of the gastrointestinal tract, respiratory tract, and skin as well other non-infectious diseases such as reproductive and psychiatric disorders. Respiratory infections were the most documented infections to be treated using plant combinations. The plant that was documented to be most commonly used in combination with other plants was Artemisia afra Jacq. ex Willd. While plant-plant combinations have drawn a marked interest, comparatively, plant-adjuvant (e.g. milk, sugar, honey, salt, vinegar, fats) combinations have attracted less research interest. Milk was reported as the most used additional ingredient in combination with medicinal plants. The combination of animal urine and dung with medicinal plants has been reported as a treatment for treat prostate infections, the human immunodeficiency virus (HIV). Other ingredients such as clay and flour were also documented, and these are often mixed with medicinal plants to treat fever, stomach ailments, sexually transmitted infections (STI) and skin conditions. Although combination therapy has been frequently reported in ethnobotanical records, over 90% of the combinations reviewed still need to be scientifically validated. CONCLUSION: Scientific reports on the antimicrobial, anti-oxidant, anti-inflammatory and other pharmacological effects of these combinations may offer an understanding of traditional combination therapy. In addition, investigation into the mechanisms of action of these combinations are also recommended to supplement the findings. Nonetheless, the use of plant combinations is still an untapped research area in southern Africa and there is a need to validate the use of those documented combinations to obtain a better understanding of combined traditional medicinal plant use.

Essential oil combinations against Clostridium perfringens and Clostridium septicum - the causative agents of gas gangrene.

AIMS: The inhibitory and bactericidal effect of a wide range of essential oils, and their selected combinations against two pathogens (Clostridium perfringens and Clostridium septicum), the causative pathogens of gas gangrenous infections were investigated. Fractional inhibitory indices were also calculated to determine the interactions. METHODS AND RESULTS: The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays were used to determine the efficacy of the essential oils. Santalum austrocaledonicum demonstrated the highest activity inhibiting both Clostridial pathogens at the lowest concentration of 0·02 mg ml-1 . Santalum austrocaledonicum combined with Cymbopogon martinii had the strongest inhibition against C. perfringens (MIC 0·02 mg ml-1 ) and C. septicum (MIC 0·01 mg ml-1 ). Selected combinations demonstrated synergy (ΣFIC ≤ 0·50) in combination against both pathogens tested. Antagonism was also observed in many combinations. CONCLUSIONS: Selected essential oils, when studied either individually or in combination, have high inhibitory and bactericidal effects against both Clostridial strains. Nine combinations have proven to be synergistic with 23 combinations additive; 96 indifferent and 77 having an antagonistic effect against the pathogenic strains. Some combinations demonstrated extreme antagonism and as such, careful consideration needs to be given to essential oil selection against these pathogens. SIGNIFICANT IMPACT OF THE STUDY: Very few essential oils have been antimicrobially screened (MIC and MBC) against Clostridial strains and furthermore, the efficacies in combination are not known.

The use of South African botanical species for the control of blood sugar.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetes mellitus (DM) is one of the most prevalent diseases globally and is of considerable concern to global health. Approximately 425 million people are estimated to have DM globally and this is predicted to increase to >642 million by 2040. Whilst the prevalence of DM in South Africa is slightly lower than the global average, it is expected to rise rapidly in future years as more South Africans adopt a high calorie "westernised" diet. Traditional medicines offer an alternative for the development of new medicines to treat DM and the usage of South African plants is relatively well documented. AIM OF THE STUDY: To critically review the literature on the anti-diabetic properties of South African plants and to document plant species used for the treatment of DM. Thereafter, a thorough examination of the related research will highlight where research is lacking in the field. MATERIALS AND METHODS: A review of published ethnobotanical books, reviews and primary scientific studies was undertaken to identify plants used to treat DM in traditional South African healing systems and to identify gaps in the published research. The study was non-biased, without taxonomic preference and included both native and introduced species. To be included, species must be recorded in the pharmacopeia of at least one South African ethnic group for the treatment of DM. RESULTS: One hundred and thirty-seven species are recorded as therapies for DM, with leaves and roots most commonly used. The activity of only 43 of these species have been verified by rigorous testing, and relatively few studies have examined the mechanism of action. CONCLUSION: Despite relatively extensive ethnobotanical records and a diverse flora, the anti-diabetic properties of South African medicinal plants is relatively poorly explored. The efficacy of most plants used traditionally to treat DM are yet to be verified and few mechanistic studies are available. Further research is required in this field.

Laboratory-based study of novel antimicrobial cold spray coatings to combat surface microbial contamination.

OBJECTIVE: To investigate the touch-contact antimicrobial efficacy of novel cold spray surface coatings composed of copper and silver metals, regard to their rate of microbial elimination. DESIGN: Antimicrobial time-kill assay. SETTING: Laboratory-based study. METHODS: An adapted time-kill assay was conducted to characterize the antimicrobial efficacy of the developed coatings. A simulated touch-contact pathogenic exposure to Gram-positive Staphylococcus aureus (ATCC 25923), Gram-negative Pseudomonas aeruginosa (ATCC 27853), and the yeast Candida albicans (ATCC 10231), as well as corresponding resistant strains of gentamicin-methicillin-resistant S. aureus (ATCC 33592), azlocillin-carbenicillin-resistant P. aeruginosa (DSM 46316), and a fluconazole-resistant C. albicans strain was undertaken. Linear regression modeling was used to deduce microbial reduction rates. RESULTS: A >7 log reduction in microbial colony forming units was achieved within minutes on surfaces with cold spray coatings compared to a single log bacterial reduction on copper metal sheets within a 3 hour contact period. Copper-coated 3-dimensional (3D) printed acrylonitrile butadiene styrene (ABS) achieved complete microbial elimination against all tested pathogens within a 15 minute exposure period. Similarly, a copper-on-copper coating achieved microbial elimination within 10 minutes and within 5 minutes with the addition of silver powder as a 5 wt% coating constituent. CONCLUSIONS: In response to the global need for alternative solutions for infection control and prevention, these effective antimicrobial surface coatings were proposed. A longitudinal study is the next step toward technology integration.

A review of the traditional use of southern African medicinal plants for the treatment of fungal skin infections.

ETHNOPHARMACOLOGICAL RELEVANCE: Human dermatophyte infections are one of the most common classes of infection globally, with an estimated 1.7 billion people contracting at least one infection annually. Southern African ethnic groups used multiple plants to treat dermatophytosis and to alleviate the symptoms, yet the anti-dermatophyte properties of most species remain poorly explored. AIM OF THE STUDY: Our study aimed to critically review the literature to document southern African plant species used to treat one or more dermatophytic infections, and to summarise scientific evaluations of these and other plant species. Our study aims to stimulate and focus future studies in this field. MATERIALS AND METHODS: A thorough review of the ethnobotanical books, reviews and primary scientific studies were undertaken to identify southern African plants used traditionally to treat dermatophytosis, thereby identifying gaps in the research requiring further study. RESULTS: Eighty-nine southern African plant species are recorded as traditional therapies for dermatophytosis. Scientific evaluations of 140 plant species were identified, although most of the species in those studies were selected for reasons apart from their traditional uses. None of those studies examined the mechanism of action of the plant species, and only a single study screened the extracts for toxicity. CONCLUSIONS: Despite southern Africa having some of the longest continuous human civilisations globally, as well as unique and diverse flora, and good ethnobotanical records, the anti-dermatophyte properties of southern African medicinal plants remains relatively poorly explored. The efficacy of the majority of plants used traditionally to treat fungal skin disease are yet to be verified and substantial further research is required in this field.

Review: Southern African medicinal plants used as blood purifiers.

ETHNOPHARMALOGICAL RELEVANCE: Blood purification practices, also referred to as blood cleansing or detoxification, is an ancient concept which is widespread amongst African traditional medicine, but for which no modern scientific basis exists. There prevails considerable ambiguity in defining what a blood purifier is. AIM OF THE STUDY: The purpose of this review is to firstly define what a blood purifier is in the context of African traditional medicine and compare to other cultural and westernized interpretations. Thereafter, this study identifies traditionally used medicinal plants used as blood purifiers in southern Africa and correlates these species to scientific studies, which may support evidence for these "blood purifying plant species". MATERIALS AND METHODS: Ethnobotanical books and review articles were used to identify medicinal plants used for blood purification. Databases such as Scopus, ScienceDirect, PubMed and Google Scholar were used to source scientific articles. An evaluation was made to try correlate traditional use to scientific value of the plant species. RESULTS: One hundred and fifty nine plant species have been documented as traditional remedies for blood purification. Most of the plant species have some pharmacological activity, however, very little link to the traditional use for blood purification. There has been some justification of the link between blood purification and the use as an antimicrobial and this has been explored in many of the plant species identified as blood purifiers. Other pharmacological studies specifically pertaining to the blood require further attention. CONCLUSION: Irrespective of the ambiguity of interpretation, medicinal plants used to "cleanse the blood", play an important holistic role in traditional medicine and this review with recommendations for further study provides some value of exploring this theme in the future.

A review of the traditional use of southern African medicinal plants for the treatment of malaria.

ETHNOPHARMALOGICAL RELEVANCE: Malaria is one of the most prevalent and deadly parasitic diseases globally, with over 200 million new cases and nearly 500,000 deaths reported annually. It is estimated that approximately half of the world's population lives in malaria endemic areas. Malaria is substantially less prevalent in South Africa than in other African regions and the disease is limited to some regions of the Limpopo, Mpumalanga and KwaZulu-Natal provinces. However, it still has a significant impact on the health of the populations living in those regions. Traditional medicines have long been used in South Africa by multiple ethic groups and many people continue to rely on these natural therapies for their healthcare. The usage of South African medicinal plants in several traditional healing systems to treat malaria have been documented (particularly for Zulu and Venda traditional medicine), although ethnobotanical investigations of other ethnic groups living in endemic malaria areas remains relatively neglected. AIM OF THE STUDY: To document the use of South African medicinal plants known to be used traditionally to treat Plasmodium spp. infections. We also critically reviewed the literature on the therapeutic properties of these and other South African plants screened against Plasmodium spp. parasites with the aim of highlighting neglected studies and fostering future research in this area. MATERIALS AND METHODS: Books and ethnobotanical reviews were examined for medicinal plants used specifically for fever. Exclusion criteria were studies not involving southern African medicinal plants. Furthermore, while fever is a common symptom of malaria, if not accompanied by the term "malaria" it was not considered. Databases including PubMed, ScienceDirect, Scopus and Google Scholar were used to source research relevant to southern African plants and malaria. Exclusion criteria were those publications where full articles could not be accessed. RESULTS: Eighty South African plant species were identified as traditional therapies for malaria. The majority of these species were documented in Zulu ethnobotanical records, despite malaria occurring in only a relatively small portion of the Zulu's traditional territory. Surprisingly, far fewer species were reported to be used by Venda, Ndebele, northern Sotho, Tsonga, Tswana, and Pedi people, despite them living in endemic malaria areas. Interestingly many of the identified species have not been investigated further. This review summarises the available ethnobotanical and laboratory research in this field, with the aim of promoting and focusing research on priority areas. CONCLUSION: Although malaria remains a serious disease affecting millions of people, medicinal plants while used extensively, have not been given the attention warranted for further investigation.

The new buzz: Investigating the antimicrobial interactions between bioactive compounds found in South African propolis.

ETHNOPHARMACOLOGICAL RELEVANCE: Propolis, a resinous substance produced by the Apis mellifera bee, contains a number of flavonoids sourced from plants found in the surrounding region. Whilst bees use this substance to seal off and protect the beehive, humans have used propolis therapeutically for centuries, making use of its antibacterial, antiseptic, antipyretic and wound healing properties, among others. South African propolis is rich in the flavonoids pinocembrin, galangin, and chrysin and very little previous research has been conducted on the antimicrobial effects of these compounds. AIM OF THE STUDY: To obtain an understanding of the antimicrobial activity of the compounds pinocembrin, galangin, and chrysin, both independently and in combination. MATERIALS AND METHODS: The compounds pinocembrin, galangin and chrysin were investigated for interactive antimicrobial activity by determining the minimum inhibitory concentrations (MIC), minimum bactericidal concentrations (MBC), anti-quorum sensing activity, biofilm studies, and toxicity studies (brine shrimp lethality assay). RESULTS: Minimum inhibitory concentration results demonstrated that combinations of compounds showed better inhibitory activity than single compounds. When the flavonoids were tested in combination using the MIC assay, synergy was noted for 22% of the 1:1 ratio combinations and for 66% of the triple 1:1:1 ratio combinations. Similarly, MBC results showed bactericidal activity from selected combinations, while the compounds on their own demonstrated no cidal activity. Quorum sensing studies showed that compound combinations are more effective at inhibiting bacterial communication than the individual compounds. Biofilm assays showed that the highest percentage inhibition was observed for the triple combination against E. coli at 24 h. Finally, brine shrimp lethality studies revealed that combinations of the three compounds had reduced cytotoxicity when compared to the individual compounds. CONCLUSION: The results obtained in this study demonstrate that the compounds found in South African propolis work synergistically to achieve an optimal antimicrobial effect, whilst simultaneously minimizing cytotoxicity.

Antimicrobial activity of Elytropappus rhinocerotis (Asteraceae) against micro-organisms associated with foot odour and skin ailments.

ETHNOPHARMACOLOGICAL RELEVANCE: The twigs of Elytropappus rhinocerotis are widely used in the Cape region of South Africa to treat foot odour, perspiration and also itchy, chilblained and burning feet. However, no antimicrobial studies have hitherto been published on this popular Cape herbal medicine, which is also used for a wide range of ailments. AIMS OF THE STUDY: To determine the antimicrobial activity of the extracts, essential oil and two major labdane diterpenes isolated from E. rhinocerotis against micro-organisms associated with foot odour and other conditions associated with skin infections. MATERIALS AND METHODS: Leafy stems were harvested from three individual plants at three separate geographical localities, giving a total of nine plant samples. The samples were air-dried, powdered and extracted with a 1:1 mixture of methanol and dichloromethane, and also with sterile distilled water. A portion of each sample was also hydrodistilled to obtain nine samples of essential oil. Isolation of the major labdane diterpenes was performed using silica and ethyl acetate in hexane (3:7 v/v) as the mobile phase. Minimum inhibitory concentrations (MIC) were determined for nine crude extracts, as well as three essential oil samples and two labdane diterpenes obtained from a bulk sample. The test organisms used in this study were from Deutsche Sammlung von Mikrooganismen (DSM) and American type culture collection (ATCC) strains and included five bacterial species (Brevibacterium agri ATCC 51663, B. epidermidis DSM 20660, B. linens DSM 20425, Staphylococcus aureus ATCC 25923, S. epidermidis ATCC 12228) and one fungal species (Trichophyton mentagrophytes ATCC 9533) associated with the skin. RESULTS: The presence of essential oil is reported for the first time. Organic extracts and essential oil samples showed moderate to noteworthy activity against selected test organisms. For the Brevibacteria, the lowest MIC values for phenolic extracts were several times lower than the corresponding values for the positive control zinc (shown in brackets): 0.0031 mg/mL against Brevibacterium agri (0.33 mg/mL), 0.17 mg/mL against B. epidermidis (3.91 mg/mL) - both for sample 2 of Vanwyksdorp; 0.13 mg/mL against B. linens (2.28 mg/mL) - for sample 3 of Vanwyksdorp. Two isolated labdane diterpenoids (one of which is here first reported) were also antimicrobially tested and showed moderate activity but had high abundance in the extracts. Two major monoterpenes and four sesquiterpenes in the essential oil were identified as 1,8-cineole (4.6-12.3%), terpinen-4-ol (9.2-24.3%), germacrene A (3.9-15.6%), (-)-spathulenol (1.7-37.8%), viridiflorol (0.3-100%), and silphiperfol-6-en-5-one (4.5-26.8%). CONCLUSION: The antimicrobial results particularly for the essential oils and against the Brevibacteria support the traditional topical use of Elytropappus rhinocerotis twigs to treat foot perspiration, foot odour and other related skin conditions.

A review of the traditional use of southern African medicinal plants for the treatment of selected parasite infections affecting humans.

ETHNOPHARMACOLOGICAL RELEVANCE: Worldwide, more than three billion cases of parasitic disease are reported yearly and it is likely that this figure is substantially under-estimated. Approximately one in six people globally are estimated to be infected with at least one parasite species annually. In South Africa, the prevalence of Schistosoma haematobium (bilharzia) and intestinal worms and helminths are particularly high, especially in children and in crowded or poorer rural communities with inadequate sanitation and nutrition. Despite alarmingly high estimates, medical research into parasitic diseases remains neglected and only malaria receives significant attention and funding. Traditional medicines have been used for centuries in Africa by multiple ethnic groups and many people rely on these healing systems as their primary healthcare modality. The traditional use of South African medicinal plants to treat parasite infestations is relatively well documented, and it is important to link these traditional uses to scientific evidence validating efficacy. AIM OF THE STUDY: To document the medicinal plants used for parasitic infections and critically review the literature on the anti-parasitic properties of South African plants against some neglected parasitic diseases. MATERIALS AND METHODS: A review of the literature (ethnobotanical books and publications documenting traditional plant use) was undertaken related to specific medicinal use for parasitic infections in Southern Africa. Inclusion criteria focused on human use. Exclusion criteria included veterinary use and malaria due to the extensive nature of these subject matters. An in-depth analysis of previous studies was undertaken and future prospectives are considered. RESULTS: In particular, bilharzia, gastrointestinal worms and helminths, ectoparasites, trichomoniasis, leishmaniasis and trypanosomiasis are reviewed with special emphasis on the gaps in research. CONCLUSIONS: Despite the availability of relatively extensive ethnobotanical records on the anti-parasitic properties of southern African medicinal plants, the antiparasitic properties of many plants have been poorly examined. There was in many instances a lack of evidence to support traditional use of many species towards some parasites and research is urgently needed in this area.

The in vitro antimicrobial evaluation of commercially essential oils and their combinations against acne.

OBJECTIVE: The study investigated the efficacy of commercial essential oil combinations against the two pathogens responsible for acne with the aim to identify synergy and favourable oils to possibly use in a blend. MATERIALS AND METHODS: Antimicrobial activity was assessed using the minimum inhibitory concentration (MIC) assay against Staphylococcus epidermidis (ATCC 2223) and Propionibacterium acnes (ATCC 11827), and the fractional inhibitory concentration index (ΣFIC) was calculated. Combinations displaying synergistic interactions were further investigated at varied ratios and the results plotted on isobolograms. RESULTS: From the 408 combinations investigated, 167 combinations were identified as displaying noteworthy antimicrobial activity (MIC value ≤1.00 mg mL-1 ). Thirteen synergistic interactions were observed against S. epidermidis, and three synergistic combinations were observed against P. acnes. It was found that not one of the synergistic interactions identified were based on the combinations recommended in the layman's aroma-therapeutic literature. Synergy was evident rather from leads based on antimicrobial activity from previous studies, thus emphasizing the importance of scientific validation. Leptospermum scoparium J.R.Forst. and G.Forst (manuka) was the essential oil mostly involved in synergistic interactions (four) against S. epidermidis. Cananga odorata (Lam.) Hook.f. and Thomson (ylang ylang) essential oil was also frequently involved in synergy where synergistic interactions could be observed against both pathogens. The combination with the lowest MIC value against both acne pathogens was Vetiveria zizanioides Stapf (vetiver) with Cinnamomum verum J.Presl (cinnamon bark) (MIC values 0.19-0.25 mg mL-1 ). Pogostemon patchouli Benth. (patchouli), V. zizanioides, C. verum and Santalum spp. (sandalwood) could be identified as the oils that contributed the most noteworthy antimicrobial activity towards the combinations. The different chemotypes of the essential oils used in the combinations predominantly resulted in similar antimicrobial activity. CONCLUSIONS: The investigated essential oil combinations resulted in at least 50% of the combinations displaying noteworthy antimicrobial activity. Most of the synergistic interactions do not necessarily correspond to the recommended layman's aroma-therapeutic literature, which highlights a need for scientific validation of essential oil antimicrobial activity. No antagonism was observed.

An in vitro investigation of indigenous South African medicinal plants used to treat oral infections.

ETHNOPHARMACOLOGICAL RELEVANCE: Over a 120 South African medicinal plants are used for the treatment of oral diseases. Despite the vast collection of antimicrobial studies being done on South African plants, there is still limited research on pathogens associated with oral infections. In consultation with the available ethnobotanical literature, this study investigates the antimicrobial efficacy of some South African medicinal plants against oral pathogens. AIM OF THE STUDY: To provide a detailed account of the antimicrobial properties of selected South African medicinal plants used traditionally to treat oral infections. The effect on Streptococcus mutans biofilm formation and the toxicity profiles of these plants are also investigated. MATERIALS AND METHODS: A total of 136 aqueous and organic extracts and six essential oils were prepared from 31 different plant species. These plant samples were screened for antimicrobial efficacy against nine oral pathogens using the micro-titre plate dilution assay. Plant extracts that were found to have noteworthy antimicrobial activity against S. mutans were further evaluated on the effect on S. mutans biofilm formation using the glass slide technique. The toxicity profiles of plant samples that were found to have noteworthy antimicrobial activity were evaluated using the brine shrimp lethality assay. RESULTS: The organic extract of Cissampelos torulosa stems displayed the lowest MIC value of 0.05mg/mL against both Lactobacillus spp. This high antimicrobial activity was also observed with the organic extract of Spirostachys africana leaves against Candida albicans. In some instances, a direct relationship was found between the traditional use of the plant and the antimicrobial activity observed. For example, noteworthy activity (MIC < 1.00mg/mL) was observed against all three Candida spp. when tested against Clematis brachiata (leaves), a plant traditionally used to treat oral thrush. Englerophytum magalismonatanum stems displayed notable activity against both Streptococcus spp. (MIC 0.83mg/mL against S. mutans and MIC 0.67mg/mL against S. sanguis). Spirostachys africana leaves displayed the greatest anti-adherent properties against S. mutans biofilm formation at both 24 and 48h, reducing the biofilm by 97.56% and 86.58% respectively. The majority of plant samples tested in the brine shrimp lethality assay (BSLA) were considered safe, however, 13 plant samples were considered toxic, at a concentration of 1mg/mL. CONCLUSION: Noteworthy antimicrobial activity for plants species such as C. brachiata and E. magalismonatnum provides validation for the traditional use of these plants. Spirostachys africana displayed the greatest reduction of adherent S. mutans cells. The BSLA results revealed that the majority of the plant samples were not toxic in nature. The findings from the results favour the potential use of these plants in treating oral diseases such as dental caries, periodontal diseases and oral thrush.

Opportunities to optimise colistin stewardship in hospitalised patients in South Africa: Results of a multisite utilisation audit.

BACKGROUND: Colistin is an old antibiotic that has been reintroduced as salvage therapy in hospitalised patients because it is frequently the only agent active against Gram-negative bacteria. Various guidelines for colistin administration have led to confusion in establishing the appropriate dose, which has potential for adverse consequences including treatment failure or toxicity. The emergence and spread of colistin resistance has been documented in South Africa (SA), but no local information exists on how and why colistin is used in hospitals, and similarly, compliance with current dosing guidelines is unknown. OBJECTIVES: To evaluate the current utilisation of colistin in SA hospitals, in order to identify stewardship opportunities that could enhance the appropriate use of this antibiotic. METHODS:  Electronic patient records of adult patients on intravenous (IV) colistin therapy for >72 hours in four private hospitals were retrospectively audited over a 10-month period (1 September 2015 - 30 June 2016). The following data were recorded: patient demographics, culture and susceptibility profiles, diagnosis, and indication for use. Compliance with six colistin process measures was audited: obtaining a culture prior to initiation, administration of a loading dose, administration of the correct loading dose, adjustments to maintenance dose according to renal function, whether colistin was administered in combination with another antibiotic, and whether de-escalation following culture and sensitivity results occurred. Outcome measures included effects on renal function, overall hospital mortality, intensive care unit length of stay (LoS), and hospital LoS. RESULTS: Records of 199 patients on IV colistin were reviewed. There was 99.0% compliance with obtaining a culture prior to antibiotic therapy, 93.5% compliance with prescription of a loading dose, and 98.5% compliance regarding prescription of colistin in combination with another agent. However, overall composite compliance with the six colistin stewardship process measures was 82.0%. Non-compliance related to inappropriate loading and maintenance doses, lack of adjustment according to renal function and lack of de-escalation following culture sensitivity was evident. Significantly shorter durations of treatment were noted in patients who received higher loading doses (p=0.040) and in those who received maintenance doses of 4.5 MU twice daily v. 3 MU three times daily (p=0.0027). In addition, compared with patients who survived, more patients who died received the 3 MU three times daily maintenance dose (p=0.0037; phi coefficient 0.26). CONCLUSIONS:  The study identified multiple stewardship opportunities to optimise colistin therapy in hospitalised patients. Urgent implementation of a stewardship bundle to improve colistin utilisation is warranted.

Antimicrobial natural product research: A review from a South African perspective for the years 2009-2016.

ETHNOPHARMACOLOGICAL RELEVANCE: This review provides information on the antimicrobial research which has taken place on South African natural products for the last eight years (2009-2016). This important field is the backbone of all studies involving the use of medicinal plants against infectious diseases and hence can form the mainstay for future studies. MATERIALS AND METHODS: All publications within the years 2009-2016 were considered. Exclusion criteria were studies not involving South African medicinal natural products and those publications where full articles could not be accessed. An overview of the most common experimental methods used and new advances in terms of antimicrobial investigations are provided. Disease categories selected for further investigation were skin and wounds, respiratory, gastrointestinal, sexually transmitted and ophthalmic infections amongst others. Alternate natural products and combinations studies were also included. RESULTS: The minimum inhibitory concentration (MIC) was the most commonly used experimental method to determine antimicrobial activity. Staphylococcus aureus was the most commonly tested skin pathogen and Klebsiella pneumoniae was the most common pathogen implicated in respiratory disorders. Only 20% of gastrointestinal studies included commonly implicated pathogens such as Shigella flexneri and Campylobacter species. CONCLUSION: Multidisciplinary studies have emerged as a strong support for antimicrobial investigations and show the importance of including toxicity when studying antimicrobial efficacy. Alternate approaches (for example biofilms and quorum sensing) at examining antimicrobial effects are encouraged. Studies on resistant strains require more insight and future recommendations should look at consistent dosing and investigations on compound interactions amongst others.

Secondary metabolites from Tetracera potatoria stem bark with anti-mycobacterial activity.

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