A Review of Ethnomedicinal Plants as Potential Anthelmintic Agents to Alternatively Control Gastrointestinal Nematodes of Ruminants in South Africa.

Small ruminant production is one of the most important animal productions for food security in the world, especially in the developing world. Gastrointestinal nematode (GIN) infection is a threat to this animal's production. Conventional drugs that are used to control these parasites are losing their efficacy due to the development of resistant parasites. These drugs are not biologically degradable, taint meat products and are also expensive for communal farmers. Hence, research is now exploring ethnomedicinal anthelmintic plants for an alternative remedy. The objective of this paper was to review ethnomedicinal plants as a potential alternative to unsustainable commercial anthelmintics. This review sought to understand common GINs infecting ruminants, resistance manifestation in GINs to conventional treatment, reasons communal farmers choose ethnomedicine, and modes of action in anthelmintic plants. It also examined the usage of plants and plant parts, dosage forms, methods for improving bioactivity, convectional validation procedures, and restrictions on ethnomedicinal plant use as anthelmintics in ethnomedicine. Such insight is essential, as it highlights the importance of ethnoveterinary medicine and ways to adopt or improve it as a potential alternative to conventional anthelmintics.

New 21-nordammarane saponins with anti-inflammatory and cytotoxic activities from Lavigeria macrocarpa Oliv.

Chemical investigation of the ethanol extract from the stems and roots of the medicinal plant Lavigeria macrocarpa led to the isolation and structure elucidation of three previously unreported 21-nordammarane-type saponins namely 6α,27-dihydroxy-3,20-dioxo-21-nordammar-24-(Z)-ene 27-O-[α-L-rhamnopyranosyl-(1→2)-ß-D-glucopyranoside] (1), 6α,27-dihydroxy-3-oxo-21-nordammar-24-(Z)-ene 27-O-ß-D-glucopyranoside (2), and 2α,3ß,6α,27-tetrahydroxy-21-nordammar-24-(Z)-ene 27-O-ß-D-glucopyranoside (3) trivially named lavigemacrocarposide A-C, along with eight known secondary metabolites. Acid hydrolysis of lavigemacrocarposide A yielded a new prosapogenin namely 6α,27-dihydroxy-3,20-dioxo-21-nordammar-24-(Z)-ene 27-O-ß-D-glucopyranoside (1a) and the previously unreported artefactual aglycones 1b and 1c. Their structures were elucidated by spectroscopic analyses including mass spectrometry, 1D and 2D NMR as well as chemical evidence. The EtOH extract, some isolated compounds as well as the prosapogenin (1a) and compounds 1b and 1c were evaluated for anti-inflammatory and cytotoxic activity. Icacine (5) exhibited a significant cytotoxicity against both HeLa and MCF-7 cell lines with an IC50 value of 0.78 µg/mL. All the tested compounds showed more that 50% inhibition of NO production, except for 1 and 2.

Leaf extract of Anacardium occidentale ameliorates biomarkers of neuroinflammation, memory loss, and neurobehavioral deficit in N(ω)-nitro-L-arginine methyl ester (L-NAME) treated rats.

PURPOSE: Anacardium occidentale commonly known as Cashew is a plant that is widely used in African traditional medicine. It is endowed with phytochemical constituents that are responsible for its medicinal properties. METHODS: Twenty-five male Wistar rats were grouped as follows: Control (Group A), Group B (L-NAME 40 mg/kg), Group C (100 mg/kg Anacardium occidentale extract plus 40 mg/kg L-NAME), Group D (200 mg/kg extract plus 40 mg/kg L-NAME) and Group E (10 mg/kg of Lisinopril plus 40 mg/kg L-NAME). The animals were treated with oral administration of either the extracts or Lisnopril daily for 4 weeks. Neuro-behavioural tests such as the Morris Water Maze and Hanging Wire Grip tests were carried out to evaluate memory/spatial learning and muscular strength, respectively. Makers of oxidative stress, antioxidant enzymes and immunohistochemical staining of Glial Fibrillary Acidic Protein and Ionised Calcium Binding Adaptor molecule 1 were assessed. RESULTS: L-NAME administration caused significant increases in biomarkers of oxidative stress, decreased antioxidant status, acetylcholinesterase activity, altered neuro-behavioural changes, astrocytosis, and microgliosis. However, Anacardium occidentale reversed exaggerated oxidative stress biomarkers and improved neuro-behavioural changes. CONCLUSIONS: Combining all, Anacardium occidentale enhanced brain antioxidant defence status, improved memory and muscular strength, thus, suggesting the neuroprotective properties of Anacardium occidentale.

L-arginine and lisinopril supplementation protects against sodium fluoride-induced nephrotoxicity and hypertension by suppressing mineralocorticoid receptor and angiotensin-converting enzyme 3 activity.

Sodium fluoride (NaF) is one of the neglected environmental toxicants that has continued to silently cause toxicity to both humans and animals. NaF is universally present in water, soil, and atmosphere. The persistent and alarming rate of increase in cardiovascular and renal diseases caused by chemicals such as NaF in mammalian tissues has led to the use of various drugs for the treatment of these diseases. The present study aimed at evaluating the renoprotective and antihypertensive effects of L-arginine against NaF-induced nephrotoxicity. Thirty male Wistar rats (150-180 g) were used in this study. The rats were randomly divided into five groups of six rats each as follows: Control, NaF (300 ppm), NaF + L-arginine (100 mg/kg), NaF + L-arginine (200 mg/kg), and NaF + lisinopril (10 mg/kg). Histopathological examination and immunohistochemistry of renal angiotensin-converting enzyme (ACE) and mineralocorticoid receptor (MCR) were performed. Markers of renal damage, oxidative stress, antioxidant defense system, and blood pressure parameters were determined. L-arginine and lisinopril significantly (P < 0.05) ameliorated the hypertensive effects of NaF. The systolic, diastolic, and mean arterial blood pressure of the treated groups were significantly (P < 0.05) reduced compared with the hypertensive group. This finding was concurrent with significantly increased serum bioavailability of nitric oxide in the hypertensive rats treated with L-arginine and lisinopril. Also, there was a significant reduction in the level of blood urea nitrogen and creatinine of hypertensive rats treated with L-arginine and lisinopril. There was a significant (P < 0.05) reduction in markers of oxidative stress such as malondialdehyde and protein carbonyl and concurrent increase in the levels of antioxidant enzymes in the kidney of hypertensive rats treated with L-arginine and lisinopril. The results of this study suggest that L-arginine and lisinopril normalized blood pressure, reduced oxidative stress, and the expression of renal ACE and mineralocorticoid receptor, and improved nitric oxide production. Thus, L-arginine holds promise as a potential therapy against hypertension and renal damage.

Naringin abrogates angiotensin-converting enzyme (ACE) activity and podocin signalling pathway in cobalt chloride-induced nephrotoxicity and hypertension.

PurposeThe persistent and alarming rates of increase in cardiovascular and renal diseases caused by chemicals such as cobalt chloride (CoCl2) in mammalian tissues have led to the use of various drugs for the treatment of these diseases. This study aims at evaluating the nephron-protective action of Naringin (NAR), a metal-chelating antioxidant against CoCl2-induced hypertension and nephrotoxicity.MethodsForty-two male Wistar rats were randomly distributed to seven rats of six groups and classified into Group A (Control), Group B (300 part per million; ppm CoCl2), Group C (300 ppm CoCl2 + 80 mg/kg NAR), Group D (300 ppm CoCl2 + 160 mg/kg NAR), Group E (80 mg/kg NAR), and Group F (160 mg/kg NAR). NAR and CoCl2 were administered via oral gavage for seven days. Biomarkers of renal damage, oxidative stress, antioxidant status, blood pressure parameters, immunohistochemistry of renal angiotensin-converting enzyme and podocin were determined.ResultsCobalt chloride intoxication precipitated hypertension, renal damage, and oxidative stress. Immunohistochemistry revealed higher expression of angiotensin-converting enzyme (ACE) and podocin in rats administered only CoCl2.ConclusionTaken together, the antioxidant and metal-chelating action of Naringin administration against cobalt chloride-induced renal damage and hypertension could be through abrogation of angiotensin-converting enzyme and podocin signalling pathway.

Anti-Inflammatory, Anti-Diabetic, Anti-Oxidant and Cytotoxicity Assays of South African Herbal Teas and Bush Tea Blends.

South Africa is home to a variety of herbal teas, such as bush tea (Athrixia phylicoides DC.), honeybush tea (Cyclopia intermedia E. Mey and C. subternata Vogel), special tea (Monsonia burkeana Planch. ex Harv.), and rooibos tea (Aspalathus linearis (Burm.f.) R. Dahlgren) that are known to possess anti-oxidant, anti-inflammatory and anti-diabetic properties. The objective of this study was to determine the in vitro anti-oxidant activity of selected tea blends using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays, as well as to assess their anti-inflammatory properties using the 15-lipoxygenase inhibitory assay. Furthermore, the study measured glucose utilisation in C2C12 myotubes. Lastly, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was used to test the safety of the tea extracts on Vero cells (African green monkey kidney cell line). Special tea and its blend with bush tea exhibited potent anti-oxidant and anti-inflammatory activity. The blending of bush tea with special tea at different ratios resulted in increased anti-oxidant activity. Although special tea had a level of cell toxicity, its toxicity was lowered during blending. All of the tea samples showed anti-diabetic effects, although with less potency as compared to insulin. The current investigation supports the use of blended herbal teas, and the positive anti-inflammatory effect of special tea warrants further research.

Lichens: An update on their ethnopharmacological uses and potential as sources of drug leads.

ETHNOPHARMACOLOGICAL RELEVANCE: Lichens, a unique symbiotic association between an alga/cyanobacterium and a fungus, produce secondary metabolites that are a promising source of novel drug leads. The beauty and importance of lichens have not been adequately explored despite their manifold biological activities such as anticancer, antimicrobial, antioxidant, anti-inflammatory, analgesic, antipyretic and antiparasitic. AIM OF THE STUDY: The present review collates and discusses the available knowledge on secondary metabolites and biological activities of lichens (in vitro and in vivo). MATERIALS AND METHODS: Using relevant keywords (lichens, secondary metabolites, bioactivity, pharmacological activities), five electronic databases, namely ScienceDirect, PubMed, Google Scholar, Scopus and Recent Literature on Lichens, were searched for past and current scientific contributions up until May 2022. Literature focusing broadly on the bioactivity of lichens including their secondary metabolites were identified and summarized. RESULTS: A total of 50 review articles and 189 research articles were searched. Information related to antioxidant, antimicrobial, anti-inflammatory, anticancer and insecticidal activities of 90 lichen species (from 13 families) and 12 isolated metabolites are reported. Over 90% of the studies comprised in vitro investigations, such as bioassays evaluating radical scavenging properties, lipid peroxidation inhibition and reducing power, cytotoxicity and antimicrobial bioassays of lichen species and constituents. In vivo studies were scarce and available only in fish and rats. Most of the studies were done by research groups in Brazil, France, Serbia, India and Turkey. There were relatively few reports from Asia and Africa despite the ubiquitous nature of lichens and the high occurrence in these continents. CONCLUSION: Secondary metabolites from lichens are worthy of further investigation in terms of their potential therapeutic applicability, including better understanding of their mechanism(s) of action. This would be of great importance in the search for novel drugs.

Clofibrate, a Peroxisome Proliferator-Activated Receptor-Alpha (PPARα) Agonist, and Its Molecular Mechanisms of Action against Sodium Fluoride-Induced Toxicity.

Sodium fluoride (NaF) is one of the neglected environmental pollutants. It is ubiquitously found in the soil, water, and environment. Interestingly, fluoride has been extensively utilized for prevention of dental caries and tartar formation, and may be added to mouthwash, mouth rinse, and toothpastes. This study is aimed at mitigating fluoride-induced hypertension and nephrotoxicity with clofibrate, a peroxisome proliferator-activated receptor-alpha (PPARα) agonist. For this study, forty male Wistar rats were used and randomly grouped into ten rats per group, control, sodium fluoride (NaF; 300 ppm) only, NaF plus clofibrate (250 mg/kg) and NaF plus lisinopril (10 mg/kg), respectively, for 7 days. The administration of NaF was by drinking water ad libitum, while clofibrate and lisinopril were administered by oral gavage. Administration of NaF induced hypertension, and was accompanied with exaggerated oxidative stress; depletion of antioxidant defence system; reduced nitric oxide production; increased systolic, diastolic and mean arterial pressure; activation of angiotensin-converting enzyme activity and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB); and testicular apoptosis. Treatment of rats with clofibrate reduced oxidative stress, improved antioxidant status, lowered high blood pressure through the inhibition of angiotensin-converting enzyme activity, mineralocorticoid receptor over-activation, and abrogated testicular apoptosis. Taken together, clofibrate could offer exceptional therapeutic benefit in mitigating toxicity associated with sodium fluoride.

The therapeutic potential of the novel angiotensin-converting enzyme 2 in the treatment of coronavirus disease-19.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of coronavirus disease 2019 (COVID-19). This virus has become a global pandemic with unprecedented mortality and morbidity along with attendant financial and economic crises. Furthermore, COVID-19 can easily be transmitted regardless of religion, race, sex, or status. Globally, high hospitalization rates of COVID-19 patients have been reported, and billions of dollars have been spent to contain the pandemic. Angiotensin-converting enzyme (ACE) 2 is a receptor of SARS-CoV-2, which has a significant role in the entry of the virus into the host cell. ACE2 is highly expressed in the type II alveolar cells of the lungs, upper esophagus, stratified epithelial cells, and other tissues in the body. The diminished expressions of ACE2 have been associated with hypertension, arteriosclerosis, heart failure, chronic kidney disease, and immune system dysregulation. Overall, the potential drug candidates that could serve as ACE2 activators or enhance the expression of ACE2 in a disease state, such as COVID-19, hold considerable promise in mitigating the COVID-19 pandemic. This study reviews the therapeutic potential and pharmacological benefits of the novel ACE2 in the management of COVID-19 using search engines, such as Google, Scopus, PubMed, and PubMed Central.

Anthelmintic, antimycobacterial, antifungal, larvicidal and cytotoxic activities of acetone leaf extracts, fractions and isolated compounds from Ptaeroxylon obliquum (Rutaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Ptaeroxylon obliquum (Thunb.) Radlk, sneezewood, is a commonly used medicinal plant in South Africa for the treatment of parasitic infections in animals, tuberculosis (TB) and related symptoms, and other microbial infections. AIM OF THE STUDY: In this study anthelmintic, antifungal, antimycobacterial, larvicidal and cytotoxic activities of the acetone leaf extract of P. obliquum, solvent-solvent derived fractions of the extract and isolated compounds (obliquumol and a mixture of lupeol and ß-amyrin) were determined. MATERIALS AND METHODS: The in vitro anthelmintic activity was tested against Haemonchus contortus ova and larvae using the egg hatch inhibition and larval development assays. The antifungal activity was investigated using a serial microplate dilution method against Aspergillus fumigatus, Cryptococcus neoformans and Candida albicans. Antimycobacterial activity was evaluated by a tetrazolium violet-based broth microdilution method against pathogenic Mycobacterium tuberculosis and Mycobacterium bovis. The larvicidal activity was evaluated against Aedes aegypti. Cytotoxicity was determined using human liver (C3A) and Vero African green monkey kidney cell lines. RESULTS: Obliquumol (ptaeroxylinol acetate) was more effective than the crude acetone extract against H. contortus ova and larvae with an LC50 of 95 µg/mL against the larvae. Obliquumol had very good antifungal activity with minimal inhibitory concentrations (MIC) values from 2 to 16 µg/mL. Lupeol and ß-amyrin mixture also had good activity against C. neoformans and C. albicans with an MIC of 16 µg/mL against both fungi. The crude extract and fractions had average to weak antimycobacterial activity against the two pathogenic mycobacteria with MICs of 313-625 µg/mL for the extract and fractions tested. Obliquumol had good activity with an MIC of 63 µg/mL against pathogenic M. tuberculosis ATCC 27115. The acetone crude extract was toxic against Vero cells with an CC50 of 14.2 µg/mL. Obliquumol and the mixture of lupeol and ß-amyrin were not toxic against both Vero and C3A cells at the highest concentration tested of 200 µg/mL and had good selectivity index values against the opportunistic fungal pathogens. No tested samples had any larvicidal activity against A. aegypti at the concentrations tested. CONCLUSION: The activity noted against H. contortus may support the ethnoveterinary use of this plant against animal parasites in South Africa although aqueous extracts were not active. The activity of obliquumol against opportunistic fungal pathogens was also promising. The cytotoxicity of the acetone leaf extract, however, raises concerns about the traditional use of P. obliquum to treat a wide range of diseases although if an aqueous extract is used the toxicity concern may not be relevant.

Potential health benefits of zinc supplementation for the management of COVID-19 pandemic.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent for the Coronavirus Disease 2019 (COVID-19). The COVID-19 pandemic has created unimaginable and unprecedented global health crisis. Since the outbreak of COVID-19, millions of dollars have been spent, hospitalization overstretched with increasing morbidity and mortality. All these have resulted in unprecedented global economic catastrophe. Several drugs and vaccines are currently being evaluated, tested, and administered in the frantic efforts to stem the dire consequences of COVID-19 with varying degrees of successes. Zinc possesses potential health benefits against COVID-19 pandemic by improving immune response, minimizing infection and inflammation, preventing lung injury, inhibiting viral replication through the interference of the viral genome transcription, protein translation, attachment, and host infectivity. However, this review focuses on the various mechanisms of action of zinc and its supplementation as adjuvant for vaccines an effective therapeutic regimen in the management of the ravaging COVID-19 pandemic. PRACTICAL APPLICATIONS: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent for the Coronavirus Disease 2019 (COVID-19), has brought unprecedented untold hardship to both developing and developed countries. The global race for vaccine development against COVID-19 continues with success in sight with attendant increasing hospitalization, morbidity, and mortality. Available drugs with anti-inflammatory actions have become alternative to stem the tide of COVID-19 with attendant global financial crises. However, Zinc is known to modulate several physiological functions including intracellular signaling, enzyme function, gustation, and olfaction, as well as reproductive, skeletal, neuronal, and cardiovascular systems. Hence, achieving a significant therapeutic approach against COVID-19 could imply the use of zinc as a supplement together with available drugs and vaccines waiting for emergency authorization to win the battle of COVID-19. Together, it becomes innovative and creative to supplement zinc with currently available drugs and vaccines.

Novel antihypertensive action of rutin is mediated via inhibition of angiotensin converting enzyme/mineralocorticoid receptor/angiotensin 2 type 1 receptor (ATR1) signaling pathways in uninephrectomized hypertensive rats.

Hypertension is the most common cardiovascular disease that affects approximately 26% of adult population, worldwide. Rutin is one of the important flavonoids that is consumed in the daily diet, and found in many food items, vegetables, and beverages. Uninephrectomy (UNX) of the left kidney was performed, followed by induction of hypertension. The rats were randomly divided into four groups of 10 rats: group 1-Sham-operated rats; group 2-UNX rats, group 3-UNX-L-NAME (40 mg/kg) plus rutin (100 mg/kg bwt), and groups 4-UNX-L-NAME plus lisinopril (10 mg/kg bwt), orally for 3 weeks. Results revealed significant heightening of arterial pressure and oxidative stress indices, while hypertensive rats treated with rutin had lower expressions of angiotensin converting enzyme (ACE) and mineralocorticoid receptor in uninephrectomized rats. Together, rutin as a novel antihypertensive flavonoid could provide an unimaginable benefits for the management of hypertension through inhibition of angiotensin converting enzyme and mineralocorticoid receptor. PRACTICAL APPLICATIONS: Hypertension has been reported to be the most common cardiovascular disease, affecting approximately 26% of the adult population worldwide with predicted prevalence to increase by 60% by 2025. Recent advances in phytomedicine have shown flavonoids to be very helpful in the treatment of many diseases. Flavonoids have been used in the treatment and management of cardiovascular diseases, obesity and hypertension. The study revealed that rutin, a known flavonoid inhibited angiotensin converting enzyme (ACE), angiotensin 2 type 1 receptor (ATR1), and mineralocorticoid receptor (MCR), comparable to the classic ACE inhibitor, Lisinopril, indicating the novel antihypertensive property of rutin. Therefore, flavonoids such as rutin found in fruits and vegetables could, therefore, serve as an antihypertensive drug regimen. Combining all, functional foods rich in flavonoids could be used as potential therapeutic candidates for managing uninephrectomized hypertensive patients.

Antihypertensive power of Naringenin is mediated via attenuation of mineralocorticoid receptor (MCR)/ angiotensin converting enzyme (ACE)/ kidney injury molecule (Kim-1) signaling pathway.

Hypertension is a condition with chronic elevation of blood pressure and a common preventable risk factor for cardiovascular disease with attendant global morbidity and mortality. The present study investigated the novel antihypertensive and neuroprotective effect of Naringenin on L-NG-Nitro arginine methyl ester (L-NAME) induced hypertension together with possible molecular mechanism of action. Rats were divided into four groups. Rats in Group A were normotensive. The hypertensive group (Group B) received 40 mg/kg) of L-NAME alone while Groups C and D were concurrently administered Naringenin (50 mg/kg) or Lisinopril (10 mg/Kg) together with L-NAME orally for 3 weeks. Blood pressure parameters, markers of oxidative stress and renal damage were measured. The immunohistochemistry of kidney injury molecule 1, mineralocorticoid receptor and angiotensin converting enzyme were also determined. Results indicated significant increases in malondialdehyde, advanced oxidation protein products, protein carbonyl contents and decrease in serum nitric oxide bioavailability in hypertensive rats. Furthermore, there were significant increases in serum myeloperoxidase, urinary creatinine, albumin and blood urea nitrogen in hypertensive rats in comparison to hypertensive rats treated with either Naringenin or Lisinopril. Immunohistochemistry reveal significant expressions of kidney injury molecule 1, mineralocorticoid receptor and angiotensin converting enzyme in hypertensive rats. However, co-treatment with either Naringenin or Lisinopril mitigated both renal and neuronal oxidative stress, normalized blood pressure and lowered the expressions of kidney injury molecule 1, mineralocorticoid receptor and angiotensin converting enzyme. Collectively, Naringenin offered a novel antihypertensive and neuroprotective effect through down regulation of kidney injury molecule 1, mineralocorticoid receptor and angiotensin converting enzyme.

Sensory Characteristics and Volatile Compounds of Herbal Teas and Mixtures of Bush Tea with Other Selected Herbal Teas of South Africa.

South Africa has a traditional heritage of using indigenous herbal teas, and the demand for herbal teas motivated by the functional health benefits has far exceeded global supply. This has led to worldwide interest in the sensory characteristics and volatile compound characterisation of herbal drink formulations. The objective of this study was to investigate the descriptive sensory analysis and volatile compounds of bush, special, honeybush and rooibos tea and the blend of bush tea with special, honeybush and rooibos, respectively. The trained sensory panel scored each tea sample for aroma, taste, aftertaste and mouthfeel attributes using sensory evaluation practices. Compound identification was performed by gas chromatography connected to a mass spectrometer (GC-MS). The results of the study demonstrated that rooibos and honeybush tea had an overall sweet-caramel, honey-sweet, perfume floral and woody aroma while bush tea and special tea depicted green-cut grass, dry green herbal and astringent/dry mouth feel. The GC-MS analyses depicted the following compounds 2-furanmethanol, 2-methoxy-4-vinylphenol, D-limonene, dihydroactinidolide, linalool, (E,E)-2,4-heptadienal, and phytol. The blending of bush tea with rooibos and honeybush tea toned down its astringent mouth feel. Compounds identified in this study may be useful markers for potential herbal tea sensory characteristics.

A comprehensive study of the potential phytomedicinal use and toxicity of invasive Tithonia species in South Africa.

BACKGROUND: Tithonia diversifolia and T. rotundifolia belong to the Asteraceae family and are native to Mexico and Central America. These plants have become invasive in parts of tropical Africa and Asia where they have become an ecological, agricultural and economic burden. Tithonia diversifolia is exploited by locals in its native and most parts of its invasive range as a source of medicines; however, T. rotundifolia is only used for medicinal purposes in one country in the native range (Venezuela) and none in the invasive range. Although T. diversifolia has been studied for different biological activities, little or no attention has been given to T. rotundifolia. This study compared the antimicrobial activity, phytochemistry, identification of bioactive compound(s) and toxicity levels of different leaf extracts and fractions of T. diversifolia and T. rotundifolia. METHODS: Antimicrobial activity was evaluated against seven pathogenic bacteria, four non-pathogenic Mycobacterium species and three fungal species using serial microdilution assays. Phytochemical contents were determined through standard methods of analysis. UPLC/MS was used to analyse the fractions to identify possible bioactive compounds that may be responsible for bioactivity, while toxicity tests were carried out using the colorimetric MTT assay and the Ames test. RESULTS: Both species had a range of antimicrobial activity against bacterial, mycobacterial and fungal species. However, T. rotundifolia displayed better activity against most of the strains tested with minimum inhibitory concentration values ranging between 0.01 and 0.07 mg/ml. Both species were rich in phenolics, flavonoids and tannins. Tagitinin A was identified as the main compound present in both species, and this compound may be responsible for the antimicrobial activity displayed. Toxicity tests showed that T. diversifolia was cytotoxic at concentrations used in this study, while T. rotundifolia was not. Both species did not show any mutagenic/genotoxic effects. CONCLUSION: The above results suggest that both species may be further developed as a source of antimicrobials for the treatment of infections caused by opportunistic pathogens. They may also serve as alternatives to highly exploited plant species with the same medicinal properties. However, T. diversifolia should be used with caution as it may be toxic.

In vitro bioassays used in evaluating plant extracts for tick repellent and acaricidal properties: A critical review.

Ticks are haematophagous arthropods which rank closely with mosquitoes in their capacity to transmit disease pathogens of importance to animals and humans. Current control of ticks is based on the routine use of synthetic chemicals administered to animals or their environment. However, years of use and overuse of these chemicals have resulted in the development of resistance in these parasites and negative environmental impacts, hence the need for cheaper, safer and more environmentally friendly alternatives with alternate modes of action. There has been a large interest in using plants for these purposes. Peer-reviewed articles on plants evaluated for their tick repellent and/or acaricidal activities against immature and adult stages of ticks were collected from nine scientific databases with the aim of reviewing the bioassays employed. Search words included "acaricidal", "tick repellent", "antitick assays" and "phytomedicine". Many different methods were used to determine repellency and acaricidal activity. These include, among a few others, petri dish, tick climbing, olfactometer, larval packet and immersion bioassays. Tick climbing repellency and adult immersion bioassays were most commonly used. Ethanol was the most widely used solvent and Rhipicephalus (Boophilus) microplus was the most commonly studied tick across all the reviewed papers. It is unclear whether the outcome of these experiments on a one-host tick can be applied to other species of ticks that infest animals and humans. Also, most of the assays on repellency did not discriminate between olfaction and tactile chemoreception-based repellency and though some of the observed methods were similar, results differ significantly. These aspects will need further evaluation. Standardized laboratory methods are required to enable the valid comparisons between results from different laboratories.

Isolation and characterization of the compounds responsible for the antimutagenic activity of Combretum microphyllum (Combretaceae) leaf extracts.

BACKGROUND: Mutations play a major role in the pathogenesis and development of several chronic degenerative diseases including cancer. It follows, therefore that antimutagenic compound may inhibit the pathological process resulting from exposure to mutagens. Investigation of the antimutagenic potential of traditional medicinal plants and compounds isolated from plant extracts provides one of the tools that can be used to identify compounds with potential cancer chemopreventive properties. The aim of this study was to isolate and characterise the compounds responsible for the antimutagenic activity of Combretum microphyllum. METHODS: The methanol leaf extract of C. microphyllum was evaluated for antimutagenicity in the Ames/microsome assay using Salmonella typhimurium TA98. TA100 and TA102. Solvent-solvent fractionation was used to partition the extracts and by using bioassay-guided fractionation, three compounds were isolated. The antimutagenic activity of the three compounds were determined in the Ames test using Salmonella typhimurium TA98, TA100 and TA102. The antioxidant activity of the three compounds were determined by the quantitative 2,2-diphenyl-1-picrylhydrazyl (DPPH)-free radical scavenging method. The cytotoxicity was determined in the MTT assay using human hepatocytes. RESULTS: A bioassay-guided fractionation of the crude extracts for antimutagenic activity led to the isolation of three compounds; n-tetracosanol, eicosanoic acid and arjunolic acid. Arjunolic acid was the most active in all three tested strains with a antimutagenicity of 42 ± 9.6%, 36 ± 1.5% and 44 ± 0.18% in S. typhimurium TA98, TA100 and TA102 respectively at the highest concentration (500 µg/ml) tested, followed by eicosanoic acid and n-tetracosanol. The antioxidant activity of the compounds were determined using the quantitative 2,2 diphenyl-1-picryhydrazyl (DPPH)-free radical scavenging method. Only arjunolic acid had pronounced antioxidant activity (measured as DPPH-free scavenging activity) with an EC50 value of 0.51 µg/ml. The cytotoxicity of the isolated compounds were determined in the MTT assay using human hepatocytes. The compounds had low cytotoxicity at the highest concentration tested with LC50 values >200 µg/ml for n-tetracosanol and eicosanoic acid and 106.39 µg/ml for arjunolic acid. CONCLUSIONS: Based on findings from this study, compounds in leaf extracts of C. microphyllum protected against 4-NQO and MMC induced mutations as evident in the Ames test. The antimutagenic activity of arjunolic acid may, at least in part, be attributed to its antioxidant activity resulting in the detoxification of reactive oxygen species produced during mutagenesis.

Phytochemical analysis with free radical scavenging, nitric oxide inhibition and antiproliferative activity of Sarcocephalus pobeguinii extracts.

BACKGROUND: Free radicals have been implicated in the pathogenesis of diverse metabolic disorders including cancer. Therefore, fighting against free radicals has become an important strategy in the prevention or treatment of such diseases, in addition to direct or indirect anticancer chemotherapy. Sarcocephalus pobeguinii has been used traditionally to treat various diseases in which excess production of free radicals is implicated, warranting investigation of its free radical scavenging, anticancer and anti-inflammatory activity. METHODS: In the present study, extracts from leaves, fruits, roots and bark of Sarcocephalus pobeguinii were evaluated on four human cancer cell lines (MCF-7, HeLa, Caco-2 and A549 cells) and a non-cancerous cell line for their antiproliferative potential. The cells were incubated with the plant extracts for 48 h at 37 °C in a 5% CO2 humidified environment and their cytotoxic effect was determined using the tetrazolium-based colorimetric (MTT) assay. The radical inhibition was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging techniques. The nitric oxide inhibitory activity was determined using LPS-activated RAW 264.7 macrophages. The correlation between radical scavenging capacity and antiproliferative activity was also analysed. RESULTS: The extract from leaves of Sarcocephalus pobeguinii (LSP) exhibited the highest cytotoxic effect on all four of the human cancer cell lines but with some cytotoxicity to the normal Vero cells. However, the LSP extract had the best selectivity index, ranging from 3.15 to 18.28. Also, antioxidant and anti-inflammatory assays indicated that the LSP extract had the highest radical scavenging capacity of all the extracts. A positive linear correlation was found between free radical scavenging ability and antiproliferative activity against the four cancer cell lines, with the highest correlation factor (R2 = 0.9914) obtained between DPPH inhibition and antiproliferative activity against A549 cells. CONCLUSIONS: The high selectivity index of the Sarcocephalus pobeguinii leaf extract indicates the potential of using this extract in cancer therapy. Furthermore, the positive correlation between free radical scavenging and antiproliferative activity suggests that the radical scavenging capacity of extracts may contribute to a prediction of their anticancer property.