Acute toxicity of the aqueous-methanolic Moringa oleifera (Lam) leaf extract on female Wistar albino rats

Background: Herbal preparations are widely assumed to be safe on oral administration and therefore the documentation of the toxic potential of some herbal concoctions used as medicine and nutrients is limited. Moringa oleifera (MO) is a plant that is gaining tremendous popularity in rural communities in Kenya as a means of offsetting nutritional and medicinal needs. However, very little is known about the safety of the plant on oral administration. Thus, the aim of the current study was to assess the biochemical and histological changes in the liver following the administration of an aqueous-methanolic (AQ-ME) MO leaf extract in female Wistar albino rats.Methods: Acute oral toxicity study on the AQ-ME MO leaf extract was conducted by the use of the limit test dose of the up and down procedure (OECD guideline number 425) with slight modifications. Briefly, ten (10) healthy, nulliparous, non-pregnant female Wistar strain albino rats aged               8-12 weeks and weighing 180±20 grams were used for the study. These animals were randomly selected into two groups; control and treatment group each having five (5) animals. They were then labelled to enable identification and control group animals were orally administered with physiological buffer saline once daily over a 48-hour period. The five (5) rats in the treatment group were dosed orally one at a time and once daily with a 2000 mg/kg dose of the AQ-ME MO leaf extract to determine the median lethal dose over a 48 hour period. Blood was then collected and used to prepare serum for biochemical analysis of aspartate amino transferase (AST), alanine amino transferase (ALT) and total bilirubin (TB) which are important biomarkers of liver dysfunction. Biochemical assays of these enzymes were performed using the method of the International Federation of Clinical Chemists (IFCC). Death was used as an endpoint, livers harvested and used to prepare transverse sections for histopathological examination. These sections were stained using the haematoxylin and eosin (H&E) method and observed for pathological changes using an optical microscope.Results: A 2000 mg/kg oral dose of AQ-ME MO leaf extract caused a significant (p0.05) increase in the mean levels of total bilirubin in the treatment group relative to the control group. On the other hand, the extract caused a non-significant (p>0.05) decrease in the mean levels of ALT in the treatment group relative to the control. The post mortem analysis of the hepatic index (liver to body weight ratio) revealed that there was a non-significant increase (p>0.05) in the hepatic index of the treatment group relative to the control. However, the transverse liver sections of treatment group animals showed mild distortions in the architecture of liver cells.Conclusions: Based on these results, the LD50 of the AQ-ME MO leaf extract was found to be >2000 mg/kg in female wistar albino rats

The legislative and regulatory framework governing herbal medicine use and practice in Kenya: a review

Complementary and alternative medicine is an integral component of primary healthcare in Kenya. This is because the infrastructural health setup in the country is inadequate in catering for all the medical needs of the population. This particularly holds true in the rural areas where many rural folk rely on products of herbal origin to offset their healthcare needs. More often than not these products are an elaborate cacophony of several different substances of biological origin and thus need personnel adept in their preparation. Sadly, due to loopholes in legislation and regulation, quacks have a field day in the practice. Moreover, the process of planting, harvesting, preparation and storage of herbs and related products dictates that a significant number of people will ultimately be involved in the whole process. This is likely to set the stage for manipulation and compromise of the safety, quality and efficacy of these products. This state of affairs appears unabated especially in the context of the current legal and regulatory framework governing herbal medicine use and practice in Kenya. Not only are these laws inadequate, they are shrouded in ambiguity, open to interpretation and the authorities mandated to implement them often end up performing duplicate roles. The aim of this review is to critique the legal and regulatory provisions governing herbal medicine use and practice in Kenya. In conclusion, laws and regulations meant to control herbal medicine use and practice in Kenya are wanting. Clear and definitive legislation on herbal medicine use and practice coupled with effective implementation by mandated institutions will go a long way in inspiring confidence to all stakeholders of herbal medicine.Keywords: Herbal medicine, legislation, regulatory framework, Keny

Exploring nature’s antidote: unveiling the inhibitory potential of selected medicinal plants from Kisumu, Kenya against venom from some snakes of medical significance in sub-Saharan Africa

Background: The present study investigated the efficacy of Conyza bonariensis, Commiphora africana, Senna obtusifolia, Warburgia ugandensis, Vernonia glabra, and Zanthoxylum usambarense against Bitis arietans venom (BAV), Naja ashei venom (NAV), and Naja subfulva venom (NSV).Methods: 40 extracts and fractions were prepared using n-hexane, dichloromethane, ethyl acetate, and methanol. In vitro efficacy against snake venom phospholipase A2 (svPLA2) was determined in 96-well microtiter and agarose-egg yolk coagulation assays. in vivo efficacy against venom-induced cytotoxicity was determined using Artemia salina. Two commercial antivenoms were used for comparison.Results: The 96-well microtiter assay revealed poor svPLA2 inhibition of BAV by antivenom (range: 20.76% ± 13.29% to 51.29% ± 3.26%) but strong inhibition (>90%) by dichloromethane and hexane fractions of C. africana, hexane and ethyl acetate extracts and fraction of W. ugandensis, dichloromethane fraction of V. glabra, and the methanol extract of S. obtusifolia. The methanol extract and fraction of C. africana, and the hexane extract of Z. usambarense strongly inhibited (>90%) svPLA2 activity in NAV. The hexane and ethyl acetate fractions of V. glabra and the dichloromethane, ethyl acetate, and methanol extracts of C. africana strongly inhibited (>90%) svPLA2 in NSV. The agarose egg yolk coagulation assay showed significant inhibition of BAV by the dichloromethane fraction of C. africana (EC50 = 3.51 ± 2.58 μg/mL), significant inhibition of NAV by the methanol fraction of C. africana (EC50 = 7.35 ± 1.800 μg/mL), and significant inhibition of NSV by the hexane extract of V. glabra (EC50 = 7.94 ± 1.50 μg/mL). All antivenoms were non-cytotoxic in A. salina but the methanol extract of C. africana and the hexane extracts of V. glabra and Z. usambarense were cytotoxic. The dichloromethane fraction of C. africana significantly neutralized BAV-induced cytotoxicity, the methanol fraction and extract of C. africana neutralized NAV-induced cytotoxicity, while the ethyl acetate extract of V. glabra significantly neutralized NSV-induced cytotoxicity. Glycosides, flavonoids, phenolics, and tannins were identified in the non-cytotoxic extracts/fractions.Conclusion: These findings validate the local use of C. africana and V. glabra in snakebite but not C. bonariensis, S. obtusifolia, W. ugandensis, and Z. usambarense. Further work is needed to isolate pure compounds from the effective plants and identify their mechanisms of action

Revered but Poorly Understood: A Case Report of Dendroaspis polylepis (Black Mamba) Envenomation in Watamu, Malindi Kenya, and a Review of the Literature

The black mamba (Dendroaspis polylepis) ranks consistently as one of the most revered snakes in sub-Saharan Africa. It has potent neurotoxic venom, and envenomation results in rapid onset and severe clinical manifestations. This report describes the clinical course and reversal of effects of black mamba envenomation in a 13-year-old boy in the Jimba area of Malindi. The victim presented to Watamu Hospital, a low resource health facility with labored breathing, frothing at the mouth, severe ptosis and pupils non-responsive to light. His blood pressure was unrecordable, heart rate was 100 beats per minute but thready, his temperature was 35.5 °C, and oxygen saturation was 83%. Management involved suction to clear salivary secretions, several hours of mechanical ventilation via ambu-bagging, oxygen saturation monitoring, and the use of South African Vaccine Producers (SAVP) polyvalent antivenom. Subcutaneous adrenaline was used to stave off anaphylaxis. The victim went into cardiac arrest on two occasions and chest compressions lasting 3–5 min was used to complement artificial ventilation. Hemodynamic instability was corrected using IV infusion of ringers lactate and normal saline (three liters over 24 h). Adequate mechanical ventilation and the use of specific antivenom remain key in the management of black mamba envenomation

Epidemiology of snake bites in selected areas of Kenya

Introduction: Snake bites are a silent public health problem in Kenya. Previous studies on snake bites in the country have mainly focused on identifying offending snake species, assessing the severity of envenomation and testing the efficacy of antivenom. Factors associated with snake bites in the country are yet to be fully understood. The aim of this work was to determine pharmaco-epidemiological factors associated with snake bites in areas of Kenya where incidence, severity and species responsible for snake bites have been reported. Methods: Kakamega provincial hospital, Kabarnet, Kapenguria and, Makueni district hospitals were selected as study sites based on previous findings on incidence, severity and species responsible for snake bites in catchment areas of these hospitals. Persistent newspaper reports of snake bites in these areas and distribution of snakes in Kenya were also considered. Cases of snake bites reported between 2007-2009 were retrospectively reviewed and data on incidence, age, site of the bites, time of bite and antivenom use was collected. Results: 176 bites were captured, 91 of which occurred in 2009. Individual incidence was between 2.7/100,000/year and 6.7/100,000/year. Bites peaked in the 1-15 year age group while 132/176 bites were in the lower limb area and 49/176 victims received antivenom. Most bites occurred during the dry season, in the bush and in the evening. Overall mortality was 2.27%. Conclusion: There is a need to sensitize the Kenyan public and healthcare personnel on preventive measures, first aid and treatment of snake bites

Acute Poisonings at a Regional Referral Hospital in Western Kenya

The emergency department (ED) of the Jaramogi Oginga Odinga Teaching and Referral Hospital (JOOTRH) handles many cases of poisoning. However, there is scant information on the factors, agents, and outcomes of poisoning at the hospital. The aim of this work was to determine the factors, agents, and outcomes of poisoning at JOOTRH. Records of patients who presented to JOOTRH with symptoms of poisoning between January 2011 and December 2016 were retrieved. Data on age, gender, offending agents, time, and season of exposure were collected. Information on the route of exposure, motive, and clinical symptoms of poisoning was also included. Other information included the laboratory evaluation, first aid measures, period of hospitalization, and outcome of poisoning. Mean, standard deviation, frequencies and bar graphs were used to describe the demographic factors of the study population. Multivariate logistic regression was used to determine the strength of association between risk factors and outcome of poisoning among patients. The level of significance for inferential analysis was set at 5%. There were 385 cases of poisoning: 57.9% (223/385) were male, 31.9% (123/385) were 13–24 years of age, and 83.9% (323/385) of exposures were in Kisumu County. The peak time of exposure was 6:00–00:00, and 23.6% (91/385) presented 1–4 h after exposure. About 62.9% (242/385) of the cases were due to accidental poisoning. Snakebites and organophosphates (OPPs) contributed to 33.0% (127/385) and 22.1% (85/385) of all cases, respectively. About 62.1% (239/385) of exposures were oral, and 63.9% (246/385) of all cases occurred in the rainy season. Additionally, 49.2% (60/122) of intentional poisoning was due to family disputes, and 16.1% (10/62) of pre-hospital first aid involved the use of tourniquets and herbal medicine. About 28.6% (110/385) of the victims were subjected to laboratory evaluation and 83.9% (323/385) were hospitalized for between 1–5 days. Other results indicated that 80.0% (308/385) responded well to therapy, while 7.3% (28/385) died, 68% (19/28) of whom were male. Furthermore, 39.3% (11/28) of the deaths were related to OPPs. Our findings suggest that the earlier the victims of poisoning get to the hospital, the more likely they are to survive after treatment is initiated. Similarly, victims of poisoning due to parental negligence are more likely to survive after treatment compared to other causes of poisoning, including family disputes, love affairs, snakebites, and psychiatric disorders. The management of JOOTRH should consider allocating resources to support the development of poison management and control

Mitigative effects of Moringa oleifera against liver injury induced by artesunate-amodiaquine antimalarial combination in wistar rats

Abstract Background Artesunate-amodiaquine (AS-AQ) is an antimalarial drug. It is associated with improved cure rates, accelerated response to therapy and delayed development of resistance. However, liver damage, neurotoxicity and agranulocytosis have been reported as adverse effects whose origins have been linked to free radicals generated by the drug. According to native materia medica, Moringa oleifera (MO) has wide utility in ethnomedicine. However, there is paucity of information on the hepatoprotective efficacy of this plant. The present study evaluated the mitigative effects of MO leaf extracts against liver injury induced by AS-AQ combination in female Wistar rats. Methods Dry leaf powder of MO was extracted with water and a 20:80 v/v mixture of water and methanol to give aqueous (AQ) and aqueous-methanol (AQ-ME) MO leaf extracts respectively. In vitro hydroxyl free radical scavenging activity of serial dilutions (10–100 μg/ml) of each of the extracts was then evaluated using an assay model where butylated hydroxytoluene (BHT) served as a reference standard. The extract with better free radical scavenging activity was then evaluated for hepatoprotective effects against AS-AQ intoxication in female Wistar rats based on the Acute Toxic Class method (OECD 2000). Serum asparate amino transferase (AST), alanine amino transferase (ALT), total bilirubin and histological examination of rat liver sections were used to evaluate the hepatoprotective activity of the selected MO leaf extract. Siliphos® (standard hepatoprotectant) was used for comparison. Results There was a concentration dependent increase in the hydroxyl free radical scavenging activity of MO leaf extracts and standard (BHT) with values ranging from 46.36–66.36% for the AQ extract, 41.04–60.95% for the AQ-ME extract and 44.93–65.23% for BHT with corresponding IC50 values of 26.84 μg/ml, 51.88 μg/ml and 32.58 μg/ml respectively. A 1000 mg/kg dose of the AQ-ME MO leaf extract significantly (p  0.05). The 1000 mg/kg dose also reduced hepatocyte degeneration in rats treated with four times the clinical dose of AS-AQ. This study suggests that the hepatoprotective activity of the leaves of MO may have some relation to its free radical scavenging properties. These leaves may thus be useful in mitigating free radical initiated disease conditions. Conclusion The aqueous-methanol Moringa oleifera leaf extract exhibits free radical scavenging and hepatoprotective properties. Further investigations on the structural identity of the phytoconstituents and their mechanisms of action should be performed to facilitate the development of a potent medicinal agent

DataSheet1_Exploring nature’s antidote: unveiling the inhibitory potential of selected medicinal plants from Kisumu, Kenya against venom from some snakes of medical significance in sub-Saharan Africa.docx

Background: The present study investigated the efficacy of Conyza bonariensis, Commiphora africana, Senna obtusifolia, Warburgia ugandensis, Vernonia glabra, and Zanthoxylum usambarense against Bitis arietans venom (BAV), Naja ashei venom (NAV), and Naja subfulva venom (NSV).Methods: 40 extracts and fractions were prepared using n-hexane, dichloromethane, ethyl acetate, and methanol. In vitro efficacy against snake venom phospholipase A2 (svPLA2) was determined in 96-well microtiter and agarose-egg yolk coagulation assays. in vivo efficacy against venom-induced cytotoxicity was determined using Artemia salina. Two commercial antivenoms were used for comparison.Results: The 96-well microtiter assay revealed poor svPLA2 inhibition of BAV by antivenom (range: 20.76% ± 13.29% to 51.29% ± 3.26%) but strong inhibition (>90%) by dichloromethane and hexane fractions of C. africana, hexane and ethyl acetate extracts and fraction of W. ugandensis, dichloromethane fraction of V. glabra, and the methanol extract of S. obtusifolia. The methanol extract and fraction of C. africana, and the hexane extract of Z. usambarense strongly inhibited (>90%) svPLA2 activity in NAV. The hexane and ethyl acetate fractions of V. glabra and the dichloromethane, ethyl acetate, and methanol extracts of C. africana strongly inhibited (>90%) svPLA2 in NSV. The agarose egg yolk coagulation assay showed significant inhibition of BAV by the dichloromethane fraction of C. africana (EC50 = 3.51 ± 2.58 μg/mL), significant inhibition of NAV by the methanol fraction of C. africana (EC50 = 7.35 ± 1.800 μg/mL), and significant inhibition of NSV by the hexane extract of V. glabra (EC50 = 7.94 ± 1.50 μg/mL). All antivenoms were non-cytotoxic in A. salina but the methanol extract of C. africana and the hexane extracts of V. glabra and Z. usambarense were cytotoxic. The dichloromethane fraction of C. africana significantly neutralized BAV-induced cytotoxicity, the methanol fraction and extract of C. africana neutralized NAV-induced cytotoxicity, while the ethyl acetate extract of V. glabra significantly neutralized NSV-induced cytotoxicity. Glycosides, flavonoids, phenolics, and tannins were identified in the non-cytotoxic extracts/fractions.Conclusion: These findings validate the local use of C. africana and V. glabra in snakebite but not C. bonariensis, S. obtusifolia, W. ugandensis, and Z. usambarense. Further work is needed to isolate pure compounds from the effective plants and identify their mechanisms of action.</p