Anti-Malarial Activity and Toxicological Effect of Combined Corymbia Citriodora, Maytenus Senegalensis and Warbugia Ugandensis as Used in Traditional Medicine in Kenya

Malaria is majorly caused by Plasmodium falciparum resulting in thousands of deaths every year. In Africa, it is a key contributor to the disease burden notable in the disability adjusted life years (DALYs). About 243 million individuals are at a risk of contracting the disease and a higher rate of deaths are observed in children under the age of five. Conventional drugs are available at a subsidized rate but the rising problem is the resistance of the plasmodium parasite to these drugs. Hence, there is an urgent need for the development of new and alternative therapeutics for treatment of malaria. In some regions in Kenya, parts of locally available plants are harvested and used for treating malaria. It is estimated that locally, 30% of the population relies on traditional medicine for treating various ailments due to the lack of infrastructure and accessible medical facilities especially in the rural areas. Warbugia ugandensis, Maytenus senegalensis and Corymbia citriodora are amongst the plants used in herbal medicine for the treatment of malaria. However, their combinatorial antimalarial efficacy and safety is yet to be determined hence the aim of this study. The plants were harvested from their natural habitats and transported to the Centre of Traditional Medicine and Drug Research (CTMDR) at the Kenya Medical Research Institute (KEMRI), Nairobi. Antimalarial properties of single and combined extracts were analyzed against Plasmodium berghei in vivo. Cytotoxic properties of the plants were carried out against the vero cell-lines in vitro by the MTT assay. Acute oral toxicity was conducted according to the OECD protocol. Effective concentration (ED50), cytotoxicity concentration (CC50) and median lethal dose (LD50) were derived. The result indicated that the combination of M. senegalensis: W. ugandensis (1:1) had the most antimalarial activity at ED50 of 1.05mg/kg whereas among the single plants W. ugandensis had the highest antimalarial activity (ED50 of 3.3mg/kg). The combinations of C. citriodora: M. senegalensis: W. ugandensis (1:1:1), C. citriodora: M. senegalensis: W. ugandensis (1:0.5:1) and C. citriodora: M. senegalensis: W. ugandensis (0.5:1:0.5) showed cytotoxicity concentration (CC50) of 101.47±3.17 µg/ml, 213.55±3.47 µg/ml and 575.80±31.40 µg/ml respectively. All the plants combinations showed no cytotoxic effects. The synergistic antimalarial properties of combined C. citriodora: M. senegalensis, C. citriodora: W. ugandensis and M. senegalensis: W. ugandensis were confirmed as the extracts showed SFIC indexes of 0.67, 0.83 and 0.28 respectively. All the plant extracts demonstrated LD50 above 2000 mg/kg with no adverse effects hence recognized as safe. This study confirms the safety and antimalarial activities of these plants and justify their use in herbal medicine practices. The results of this study sets the precedence for the development of an antimalarial herbal formulation that is less toxic and more affordable.Keywords: Anti-Malarial Activity, Toxicological Effect, Traditional Medicine DOI: 10.7176/JNSR/12-22-04 Publication date: November 30th 202

Effects of methanol and aqueous extracts of Acacia xanthophloea, Strychnos heninningsii and Microglossa pyrifolia on Immunoglobin E using asthma induced mice model

Background: The increasing prevalence of asthma in developing countries during the last decade continues to represent a significant public health problem, causing both economic and social burdens. It remains an area of considerable unmet medical need which affects 235–330 million and kills about 300,000 people worldwide. Low and middle income countries make up more than 80% of the mortality and the prevalence of Asthma in Kenya is 15.8%.Treatment of asthma still remains far from being satisfactory, which is severely limited by undesirable adverse effect and high cost. Methanol and aqueous extracts of Acacia xanthophloea, Strychnos heninningsii and Microglossa pyrifolia have shown efficacy on antimicrobial and antioxidant properties but have not been investigated for anti-asthmatic activities. Objectives: This study was aimed at evaluating the anti-asthmatic activities of extracts of Acacia xanthophloea, Strychnos heninningsii and Microglossa pyrifolia on asthma induced mice. Materials and Methods: Female Swiss Albino mice aged 8weeks old and weighing 20 +/- 2g, were asthma induced by using 1% Ovalbumin (grade VI; Sigma, Steinheim, Germany) followed by treatment using methanol and water extracts of A xanthophloea, S heninningsii and M pyrifolia in concentrations of 50, 100 and 200mg/kg body weight except for positive control group of mice which was induced and not treated. Standard reference drug control group was given 10mg/kg Prednisolone. After treatment, serum total Immunoglobin E (IgE) levels were determine using mouse OVA specific IgE Enzyme Linked Immunosorbent Assay (ELISA) (BioLegend).Cytotoxicity screening of the plant extracts was also done using Vero E6 cells and MTT dye. Data were analyzed and expressed as Means and Standard Deviation and the parametric data was statistically analyzed using one way Analysis of Variance (ANOVA) followed with unpaired student’s t-test at p-value < 0.001. Results: The results showed that the extracts were able to reduce the serum total IgE levels by upto100% in reference to the positive control. Conclusion: The extracts tested have the ability to reduce IgE levels in an asthmatic attack. The results can to be used for future possible large scale implementation in an effort to solve the burden of asthma as well as the current anti-asthmatic drug side effects. Keywords: Anti-asthma, Immunoglobin E, Acacia. xanthophloea, Strychnos heninningsii, Microglossa pyrifoli

Pyrethrins In Soil and Water From Selected Pyrethrum Growing Areas In Nakuru County, Kenya

Introduction: Pyrethrum also known as Chrysanthemum is a plant from which compounds known as pyrethrins are derived. The pyrethrins have  been used for many years as insecticides. Incidentally due to their high instability they have slowly been replaced by synthetic pyrethroids.  Pyrethrins are generally regarded as safe compared to the pyrethroids. However, the amounts released into the environment have not been well documented especially in pyrethrum growing regions.Objective:  The aim of the study was to determine the concentration of pyrethrins that come from pyrethrum plants and released into the  environment through their use as insecticides, thus, into drinking water and soil, in pyrethrum growing regions in Kenya.Methodology: Quantification to amounts of pyrethrins from pyrethrum plants, in soil and water bodies in and around pyrethrum farms in Kiambogo and Naivasha (Nakuru County). The study was carried out using High Performance Liquid Chromatography (HPLC). Water samples (0.5L) were collected from the following water bodies: rivers, streams, dams, wells and boreholes near or within pyrethrum farms.Conclusion: It was established that, the quantity of pyrethrins present in water and soil samples werebelow detectable levels within the WHO recommended range. Hence safe for the environment,more so for the farmers and the people living around pyrethrum farms. Key words: Pyrethrum, pyrethrins, chromatography, water, soil

Validation of Safety and Efficacy of Antitussive Herbal Formulations

Background: Cough is an important defensive pulmonary reflex that removes irritants, fluids or foreign materials from the airways. Frequently, cough is non-productive and requires suppression and opioid receptor agonists such as codeine are commonly used as antitussive agents. However, opioids produce side effects that include sedation, addiction potential and constipation. Novel cough suppressant therapies should maintain or improve upon the antitussive efficacy profile of opioids but with minimum or no side effects. Objective: To evaluate antitussive activity of combination of herbal medicines as formulations in sulphur dioxide - induced cough model in rats. Methodology: Wister rats of either sex, weighing 150 - 200 g, were divided into 7 groups (n = 6). Group 1 served as a control and received normal saline, groups 2 received codeine phosphate, group 3 and 4 received the coded market samples and groups 5, 6 and 7 received the test samples, respectively. Thirty or sixty minutes following administration, the rats were exposed to sulphur dioxide gas for 1 minute and then placed in an open chamber for counting of cough bouts. Results: The formulations exhibited cough inhibitions of between 15 and 27%, and 14 and 38%, with respect to the control group, 30 and 60 minutes after sample administration respectively. Conclusion: The herbal formulations demonstrated significant (p < 0.05) antitussive activity in sulphur dioxide induced cough model. Key words: Antitussive activity; herbal formulations; sulphur dioxide; coug

Development of a nutraceutical from natural products: A case study of a herbal-based low sodium table salt

Background: Cyperus papyrus reed ash has been used traditionally as a salt substitute in Western Kenya. Previous work carried out at Kenya Medical Research Institute indicated that potassium salt substitution derived from local papyrus reed has a favourable potassium/sodium ratio that is suitable for use to regulate high blood pressure. Objective: To develop and design a process flow for pilot scale production and to develop suitable analytical methods for quality assurance for a herbal based low sodium table salt. Methodology: The plant material was collected from two study sites in Rift Valley then cleaned, chopped, dried, ashed and extracted at laboratory and optimized industrial scale to yield laboratory and pilot scale samples. Elemental analysis was determined using Atomic Absorption and Flame emission spectroscopy. Results: The herbal salt yield for the laboratory scale processing was about 10% for both samples but 13% and 22%, respectively, using the optimized industrial procedure. Elemental analysis results indicate the presence of both essential and non-essential elements and heavy metal was within the World Health Organization acceptable limits. The potassium/sodium ratios obtained were between 3 and 11. Discussion: The herbal salt has the capacity to preserve meat and is a source of other essential trace elements such as chromium, Zinc and manganese. Key words: Nutraceuticals; process optimization; low sodium; herbal salt; elemental compositio

In vitro anti-viral activity of aqueous extracts of Kenyan Carissa edulis , Prunus africana and Melia azedarach against human cytomegalovirus

The aqueous extracts of three medicinal plants, Carissa edulis (Forssk.) Vahl (Apocynaceae), Prunus africana (Hook.f.) Kalkm (Rosaceae) and Melia azedarach L. (Meliaceae) have shown significant reduction in the replication of human cytomegalovirus (HCMV) in human embryonic lung (HEL) fibroblasts cells in vitro. Using the plaque inhibition assay for the determination of anti-viral activity, the HEL fibroblast cells cultured in 24 well plates were infected with 1 x 102 PFU 91S HCMV and treated with various concentrations of the extracts. The plaques formed were counted after 7 days incubation at 37\ub0C in 5% CO2 and the percent plaques inhibited were calculated against infected untreated control. The effective concentrations inhibiting plaque formation by 50% (EC50) was found between 40 to 80 \u3bcg/ml for all the extracts. The cell cytotoxic concentrations (CC50) for each of the three extracts, by the trypan blue exclusion test, gave a safe therapeutic index. These results have demonstrated the potential anti-viral activities of the extracts of the three medicinal plants at non-cytotoxic concentrations

Secondary bacterial infections and antibiotic resistance among tungiasis patients in Western, Kenya.

Tungiasis or jigger infestation is a parasitic disease caused by the female sand flea Tunga penetrans. Secondary infection of the lesions caused by this flea is common in endemic communities. This study sought to shed light on the bacterial pathogens causing secondary infections in tungiasis lesions and their susceptibility profiles to commonly prescribed antibiotics. Participants were recruited with the help of Community Health Workers. Swabs were taken from lesions which showed signs of secondary infection. Identification of suspected bacteria colonies was done by colony morphology, Gram staining, and biochemical tests. The Kirby Bauer disc diffusion test was used to determine the drug susceptibility profiles. Out of 37 participants, from whom swabs were collected, specimen were positive in 29 and 8 had no growth. From these, 10 different strains of bacteria were isolated. Two were Gram positive bacteria and they were, Staphylococcus epidermidis (38.3%) and Staphylococcus aureus (21.3%). Eight were Gram negative namely Enterobacter cloacae (8.5%), Proteus species (8.5%), Klebsiellla species (6.4%), Aeromonas sobria (4.3%), Citrobacter species (4.3%), Proteus mirabillis(4.3%), Enterobacter amnigenus (2.1%) and Klebsiella pneumoniae (2.1%). The methicillin resistant S. aureus (MRSA) isolated were also resistant to clindamycin, kanamycin, erythromycin, nalidixic acid, trimethorprim sulfamethoxazole and tetracycline. All the Gram negative and Gram positive bacteria isolates were sensitive to gentamicin and norfloxacin drugs. Results from this study confirms the presence of resistant bacteria in tungiasis lesions hence highlighting the significance of secondary infection of the lesions in endemic communties. This therefore suggests that antimicrobial susceptibility testing may be considered to guide in identification of appropriate antibiotics and treatment therapy among tungiasis patients

In vitro anti-viral activity of aqueous extracts of Kenyan Carissa edulis , Prunus africana and Melia azedarach against human cytomegalovirus

The aqueous extracts of three medicinal plants, Carissa edulis (Forssk.) Vahl (Apocynaceae), Prunus africana (Hook.f.) Kalkm (Rosaceae) and Melia azedarach L. (Meliaceae) have shown significant reduction in the replication of human cytomegalovirus (HCMV) in human embryonic lung (HEL) fibroblasts cells in vitro. Using the plaque inhibition assay for the determination of anti-viral activity, the HEL fibroblast cells cultured in 24 well plates were infected with 1 x 102 PFU 91S HCMV and treated with various concentrations of the extracts. The plaques formed were counted after 7 days incubation at 37°C in 5% CO2 and the percent plaques inhibited were calculated against infected untreated control. The effective concentrations inhibiting plaque formation by 50% (EC50) was found between 40 to 80 μg/ml for all the extracts. The cell cytotoxic concentrations (CC50) for each of the three extracts, by the trypan blue exclusion test, gave a safe therapeutic index. These results have demonstrated the potential anti-viral activities of the extracts of the three medicinal plants at non-cytotoxic concentrations

Evaluation of the HIV-1 reverse transcriptase inhibitory properties of extracts from some medicinal plants in Kenya

Extracts from twenty two medicinal plants popularly used in preparing traditional remedies in Kenya were screened for activity against the HIV-1 reverse transcriptase. The screening procedure involved the use of tritium labeled thymidine triphosphate as the enzyme substrate and polyadenylic acid.oligodeoxythymidylic acid [poly(rA).p(dT)12-18] as the template primer dimer. Foscarnet was used as a positive control in these experiments. At a concentration of 100µg/ml, extracts from eight of these plants showed at least 50 per cent reverse transcriptase inhibition. This activity was abitrarily considered as significant. This indicates that there is the probability that some antiretroviral compounds could be identified and isolated from materials from these plants. [Afr. J. Health Sci. 2002; 9:81-90

Traditional, complementary and integrative healthcare: global stakeholder perspective on WHO’s current and future strategy

As the ‘WHO Traditional Medicine Strategy: 2014–2023’ is entering its final phase, reflection is warranted on progress and the focus for a new strategy. We used WHO documentation to analyse progress across the objectives of the current strategy, adding the role of traditional, complementary and integrative healthcare (TCIH) to address specific diseases as a dimension absent in the current strategy. Our analysis concludes on five areas. First, TCIH research is increasing but is not commensurate with TCIH use. TCIH research needs prioritisation and increased funding in national research policies and programmes. Second, WHO guidance for training and practice provides useful minimum standards but regulation of TCIH practitioners also need to reflect the different nature of formal and informal practices. Third, there has been progress in the regulation of herbal medicines but TCIH products of other origin still need addressing. A risk-based regulatory approach for the full-range of TCIH products seems appropriate and WHO should provide guidance in this regard. Fourth, the potential of TCIH to help address specific diseases is often overlooked. The development of disease strategies would benefit from considering the evidence and inclusion of TCIH practices, as appropriate. Fifth, inclusion of TCIH in national health policies differs between countries, with some integrating TCIH practices and others seeking to restrict them. We encourage a positive framework in all countries that enshrines the role of TCIH in the achievement of universal health coverage. Finally, we encourage seeking the input of stakeholders in the development of the new WHO Traditional Medicine Strategy