Indian Journal of Traditional Knowledge Vol. 17(4), October 2018, pp. 716-723

In vivo antiplasmodial activities of four Nigerian used singly and in polyherbal combination against Plasmodium berghei infection

Orabueze Celestina Ia*, Adesegun Sunday Aa, Ota Duncan Ac & Coker Herbert Ab aDepartment of Pharmacognosy University of Lagos, PMB 12003, Surulere, Lagos, ; bDepartment of Pharmaceutical Chemistry, Faculty of Pharmacy, cDepartment of Physiology, College of Medicine, University of Lagos, PMB 12003, Surulere, Lagos, Nigeria E-mail: [email protected]

Received 03 October 2017, revised 14 August 2018

Methanolic extracts from 4 medicinal plants representing 4 families, used traditionally for malaria treatment in South east Nigeria were screened for their in vivo antimalarial activity in mice against a chloroquine (CQ)-sensitive Plasmodium berghei NK65, alone and in combination as polyherbal remedy. The methanolic extracts of individual plants in single and in combination (100-400 mg kg-1) were administered orally to P. berghei-infected mice in both early and established models of antiplasmodial studies. Survival time was determined. When used alone, extracts from the 4 plants, Fadogia cienkowskii (FC), Lophira lanceolata (LL), Vernonia conferta (VC) and madiensis (PM) had statistically significant parasitaemia suppression (62.06 – 93.44 %) and curative (48.93 – 72.47 %) effects. Lower doses of the 4 individual plants constituted FLVP at a combination ratio of 1: 1: 1: 1. Polyherbal formulation (FLVP) gave statistically significant suppression and curative which ranged from 45.5 – 85.1 % and 45.5 – 74.00 %, respectively. A more general improved antimalarial recovery effect, controlled weight lost and enhanced survival rate of the test mice compared to the individual therapeutic effect was observed. The standard drug, CQ gave stronger curative effect 100 % parasitaemia clearance. Our study findings suggest that the 4 plants used both as monotherapy and combined polyherbal remedy showed antiplasmodial in vivo activities and FLVP showed a more stable recovery status. FLVP is safe up to tested dose of 4000 mg kg-1. Further studies using varying fixed ratios for FLVP could result in better and improved antimalarial formulation.

Keywords: Antiplasmodial, Curative activity, Suppressive activity, Polyherbal formulation, Parasitaemia, Synergistic IPC int. Cl.8: A61K 36/00, A01D 20/46, A61K 39/00, A61K 31/47, A61K 31/4706, A61K 31/357

In Nigeria, malaria is one of the most prevalent keeping infection and re-infection of malaria under infections and constitutes one of the main causes of control but that may be in the near future since death particular in children under the age of 5 yrs, effective vaccine is still not available. At present the pregnant women and immunity challenged adults1,2,3. clinical strategy for malaria therapy concentrates in Malaria is associated with many complications which use of combination therapy as advised by WHO, include severe anemia, cerebral malaria, respiratory while focusing on research for new and more distress syndrome, and low birth weight, which effective drugs due to the continuous development of contribute to final cause of death4. World Health resistance by the causative agent to available drugs. Organization (WHO) records showed that about an Medicinal plants consist of many bioactive estimated 303 000 under fives globally, including phytoconstituents, exhibiting different pharmacological 292 000 in the African succumbed to malarial activities, mechanisms of actions and biochemical associated death in the year 20155. Efforts to path ways7,8. The health benefit or risk may progressively reduce the high malaria mortality and increase when a combination of medicinal plants is morbidity rates is challenged by the ease at which the presented as poly herbal formulation. Fixed dose parasite, particularly Plasmodium falciparum develop formulations or drug combination is currently resistance to available and inexpensive conventional preferred over a single drug in many multidrug drugs and vectors to insecticides3,6. Preventive effect resistant infections such as bacterial infections, of vaccine would have been a better approach towards malaria, tuberculosis and HIV9 because of increased ——————— efficacy. Development of parasite resistance to *Corresponding author artemisinin and its combinations is being reported ORABUEZE et al.: ANTIMALARIAL POTENTIAL OF NIGERIAN POLY HERBAL COMBINATION 717

in some parts of the world and there is a need for Experimental animals more treatment options10. Swiss albino mice weighing between 18-22 g were A large number of medicinal plants have been obtained from the College of medicine animal care reported as potential sources of antiplasmodial agents. center, Department of Physiology, College of These plants are used as single or as concoction of two Medicine of the University of Lagos. The animals or more plants. A polyherbal concoction of four (4) were accommodated in a clean, well ventilated but medicinal plants namely, Lophira lanceolate, Protea grouped caging system. Suitable nutritional, madiensis, Vernonia conferta and Fadogia cienkowskii uncontaminated food and water was provided at all is used in South east of Nigeria as antimalarial remedy. times and their beddings frequently changed. The Lophira lanceolata Tiegh, is a medicinal plant found in animals were allowed to acclimatize 7 days preceding , including Nigeria. It’s has been the experiment. Dark and light cycles were documented to exhibit antimalarial potentials. Other maintained at 12 h each. The animals were used in reported uses of the plant include pain management, accordance with the guideline and recommendation of intestinal troubles, fever, respiratory tract infections, the ethics committee on the use of animals for dysentery, antidiarrheal and spasmolytic properties11,12. research of the University of Lagos ethical committee, The stem bark of Protea madiensis Oliv, is used in the Lagos, Nigeria. treatment of diarrhoea, amenorrhoea, malaria, fever, headache and dysentery13,14. Vernonia conferta Benth., Malaria parasite belongs to the family Asteraceae. In traditional The parasite, chloroquine sensitive Plasmodium medicine, the use of Vernonia species include, berghei (NK-65 strain) was obtained from National anti-microbial, anti-malarial, analgesics, stomachache, Institute of Medical Research (NIMR) Yaba Lagos anti-inflammatory, cytotoxic, anti-oxidant, worm Nigeria. It was then maintained in University Animal expellant, various gastro-intestinal disorders, jaundice, House Laboratory by serial passage of the parasite in and hypolipidemic effects15-17. Fadogia cienkowskii an uninfected mice.

Schweinf (Rubiaceae), locally called ‘ufu-ewureje’ in Preparation of plant sample Nigeria is used in relief of headache, malaria, general The collected samples (4 plants) were cleaned and body debility, inflammation, diarrhoea and 18,19 dried at room temperature for two weeks. The dry impotence . plant samples were ground into powder using pestle This study was carried out to evaluate the antimalarial and mortar. Each of the powders obtained was used to effects of these plants used singly as monotherapy and prepare individual extract. Cold maceration technique when formulated as polyherbal product. was employed for extraction, using 3600 mL of 90 % methanol and 400 g of each plant materials. The Materials and methods combination extract (FLVP) was prepared by Equipment/reagents cold maceration of the 4 plant raw materials at the Giemsa stain (Sigma-Andrich chemical company, ratio of 100: 100: 100: 100 mg. The filtrates were St. Louis, USA), Chloroquine phosphate, DMSO concentrated under vacuum using a rotary evaporator (Dimethyl sulphate), methanol and other chemicals and (Buchi Rota vapor, Germany) at 45 rpm and 40 oC to reagents used for this study were of analytical grade. obtain the crude extracts. The dried extracts were o Plant samples collection and identification stored in a refrigerator at 4 C in air tight containers Leaves of the plants were collected at until used.

Nsukka, Enugu state in February 2016. The Phytochemical analysis of the crude extract plants were identified and authenticated by a A preliminary phytochemical study of the 4 plant taxonomist, Mr. Ozioko Fred, from the International extracts were performed using standard procedures. center for ethnomedicine and drug development (INTERCEDD), Nsukka Enugu state. The herbarium Determination of acute toxicity of crude extract of specimens were deposited and the following the polyherbal combination, FLVP voucher numbers assigned: Lophira lanceolate The median lethal dose (LD50) was determined for (INTERCEDD/005), Protea madiensis (INTERCEDD/ estimating acute toxicity of the crude extract of the 010), Vernonia conferta (INTERCEDD/1609) and polyherbal formulation, FLVP in Swiss albino mice Fadogia cienkowskii (INTERCEDD/1607). model using Lorke method (1983)20. This involved 718 INDIAN J TRADIT KNOWLE, VOL. 17, NO. 4, OCTOBER 2018

oral administration of 3 different doses of the extract Eighty five mice were intraperitoneally inoculated (1000, 2000 and 4000 mg kg-1) to groups of 6 mice with standard inoculums of 1x107 P. berghei each. The animals were observed for manifestation of NK-65 infected erythrocytes on the first day physical signs of distress and toxicity such as known as Day 0 (D0). After 72 h (D3) and writhing, decreased motor activity, stressed confirmation of parasitaemia, the mice were respiration and death for a period of 7 days. randomly divided into groups (Table 1) of 5 mice each and dosed accordingly. Doses of 100, 200 and Antimalarial activity of the extracts 400 mg kg-1 of each extract were given accordingly

daily for 5 consecutive days (D3 – D7), control Animal grouping -1 Animals were grouped according to extract and groups received 10 mg kg chloroquine and 0.2 mL dose level (Table 1). of 5 % DMSO. Geimsa stained thin blood film was prepared from the tail of each animal daily for Chemo-suppression test the 5 dosing days (D3 –D7) and D8 (24 h post last A 4-day suppressive test on early infection was drug administration) and examined microscopically 21 used for the evaluation as described by Peter et al. . to monitor parasitaemia level. The prepared slides The animals were infected intraperitoneally with were viewed under X 100 objective (oil immersion) 7 blood containing 1 x 10 parasitized red blood cells in light microscope. The number of deaths that 0.2 mL inoculum on first day (D0). They were occurred in each group over a period of 30 days randomly divided into test groups of 5 mice each. (D0 - D29) were noted and mean survival time (MST) Two hours post infection, all the test groups (Table 1) for each group determined. of the animals were orally administered with the extracts accordingly. Three different dose levels were Statistical analysis -1 used, 100, 200 and 400 mg kg for each extract. Two The analysis was carried out in n = 5 for all (2) test groups of the animals received the reference determinations and the results were expressed as -1 drugs (chloroquine 10 mg kg and artesunate 15 mg mean ± SEM. The significance of difference between -1 kg ) while the last group received the vehicle, 5 % the controls and treated groups were determined using DMSO at 0.2 mL. The treatment was repeated daily one-way analysis of variance (ANOVA). p < 0.05 was for 4 consecutive days (D0 - D3). On the fifth day (D4) considered to be statistically significant. post-inoculation, smears were made using veno- punction (cutting the tip of the tail to get drops of Results blood directly to the glass slide). The % suppression of parasitaemia was calculated by comparing the Phytochemical test parasitaemia present in infected negative control with Results obtained from phytochemical screening of those of test mice. the extracts showed some secondary metabolites The body weights of the experimental animals (Table 2). were measured before parasitizing (D ) them with 0 Acute toxicity test P. berghei and before blood collection (D4) for smear. There was no mortality recorded in the mice Curative model upon oral administration even at 4 000 mg kg-1. Evaluation of curative potential of the extracts This indicates that the experimental doses used are were carried out as described by Iyiola et al., 201122. relatively safe.

Table 1 — Animal grouping Extract/Drug 100 mg kg-1 200 mg kg-1 400 mg kg-1 Others mg kg-1 Negative control - - - 0.2 mL Fadogia cienkowskii FC FC FC - Lophira lanceolate LL LL LL - Vernonia conferta VC VC VC - Protea madiensis PM PM PM - Polyherbal FLVP FLVP FLVP FLVP - Chloroquine - - - 10 Artesunate - - - 15 ORABUEZE et al.: ANTIMALARIAL POTENTIAL OF NIGERIAN POLY HERBAL COMBINATION 719

Antimalarial test of 93.44 %, the standard drugs CQ (10 mg kg-1) and Art (15 mg kg-1) caused 89.36 and 85.5 % Chemosuppressive effects of methanolic extracts suppression, respectively. Chemosuppressive activity compared in malaria induced mice with P. berghei of CQ was significantly less than the effect of All the extracts showed dose-dependent -1 400 mg kg of FC but stronger than those of the schizonticidal effects on the parasitaemia at the doses other 3 individual test extracts and extract of employed in the study (Table 3). These effects were polyherbal combination (FLVP) of the 4 plants. statistically significant relative to the negative control

(p < 0.05). The chemosuppressive potential of FLVP Antiplasmodial effect of methanol extracts in singles is lower than that of Fadogia cienwoskii (FC) and and in polyherbal combination on established standard drugs (CQ and Art). Fadogia cienwoskii infection (FC) at 400 mg kg-1 has the highest chemosuppression The extracts showed dose-dependent curative Table 2 — Phytochemical constituents of plant extracts effect on the parasitaemia (Table 4). The percentage parasitaemia reductions observed in all the various TEST Fadogia Vernonia Lophira Protea cienwkoskii conferta lanceolata madiensis extract treated groups were statistically significant Alkaloids + + + + relative to the negative control (p < 0.05). The Phenols + - - + standard drug (CQ) used gave a total clearance Flavonoid + + + + (100 %) of the parasites by the last day of the Saponins + - - - treatment, followed by the FLVP at 74.00 % and Sterols & + + + - FC with 72.47 % parasitaemia clearance. The survival triterpenes time of animals were improved particularly with Glycosides + - - - CQ, FLVP and FC (Table 5). Deoxy + + + - sugars Discussion Lactone - - - - The use of polyherbal remedy is a commonly ring employed procedure by traditional healers in Africa. + and - means presence and absence respectively Table 3 — Suppressive effect of control and extracts against P. berghei infection in mice

-1 Drug Dose mg kg Weight D0 Weight D4 % Weight change % Parasitaemia % Chemosuppression 5% DMSO 0.2 ml 20.17 ± 0.63 18.03 ± 0.13 -10.61 11.28±0.35 0 CQ 10 20.80 ± 0.50 21.10 ± 0.23 1.44 1.20 ± 0.02* 89.36 Art 15 20.07 ± 0.36 20.26 ± 0.61 0.95 1.64 ± 0.45* 85.46 FLVP 100 20.20 ± 0.79 19.81 ± 1.24 -1.93 3.083 ± 0.23* 72.7 200 19.42 ± 0.51 19.70 ± 0.63 1.44 2.633 ± 0.08* 76.68 400 20.19 ± 0.92 21.34 ± 0.44 5.70 1.518 ± 0.18* 86.52 PM 100 20.14 ± 0.83 19.46 ± 0.58 -3.38 4.283 ± 0.37* 62.06 200 19.40 ± 0.16 18.80 ± 0.62 -3.09 3.408 ± 0.16* 69.77 400 20.04 ± 0.27 20.10 ± 0.32 0.30 2.483 ± 0.17* 78.01 VC 100 18.80 ± 0.23 18.67 ± 0.43 -0.69 4.085 ± 0.20* 63.74 200 19.74 ± 0.62 19.30 ± 0.30 -2.23 3.573 ± 0.18* 68.35 400 21.02 ± 1.10 21.04 ± 1.02 0.10 3.16 ± 0.23* 71.99 FC 100 19.82 ± 0.45 20.16 ± 0.45 1.72 2.05 ± 0.06* 81.82 200 21.40 ± 0.45 21.87 ± 0.45 2.20 1.458 ± 0.11* 87.06 400 20.51 ± 0.45 21.36 ± 0.45 4.144 0.736 ± 0.10* 93.44 LL 100 19.30 ± 0.45 18.78 ± 0.45 -2.69 3.453 ± 0.08* 69.41 200 20.00 ± 0.45 20.02 ± 0.45 0.1 2.705 ± 0.11* 75.98 400 18.43 ± 0.45 18.91 ± 0.45 2.60 2.083 ± 0.17* 81.56 N = 5; *: Significance at p < 0.05; CQ: Chloroquine

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Table 4 — Curative effect of individual and combined extracts on P. berghei in mice Drug Dose mg kg-1 Parasitaemia Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 5% DMSO 100 % Parasitaemia 11.31± 0.28 13.49%±1.04 15.38±0.64 18.30±0.42 20.47±0.34 16.31±0.51 % Cure 0 0 0 0 0 0

CQ 5 % Parasitaemia 11.66±0.32 10.70±0.84 4.38±0.68 1.39±0.24 0±0 0±0 % Cure -3.09 20.68 71.52 92.40 100 100

FLVP 100 % Parasitaemia 11.36 ±0.61 9.53 ±0.67 10.63 ±0.47 9.56 ±0.70 7.64 ±0.46 5.43 ±0.38 % Cure -0.44 29.36 30.88 47.76 62.68 66.71 200 % Parasitaemia 11.52 ±0.19 12.55 ±0.19 11.82 ±0.40 9.26 ±0.34 7.51 ±0.52 5.59 ±0.46 % Cure -1.86 6.97 23.15 49.40 63.31 65.73 400 % Parasitaemia 10.82 ±0.47 11.61 ±0.17 10.00 ±0.17 8.67 ±0.47 5.94 ±0.23 4.24 ±0.30 % Cure 4.33 13.94 34.98 52.62 70.98 74.00

PM 100 % Parasitaemia 11.12±0.47 11.12±0.47 10.03±0.32 9.00±0.19 7.44±0.24 6.59±0.24 % Cure 1.68 1.68 34.79 50.82 63.65 59.60 200 % Parasitaemia 11.36±0.36 11.36±0.36 10.12±0.20 8.89±0.16 7.79±0.22 6.57±0.41 % Cure -0.44 -0.44 34.20 51.42 61.94 59.72 400 % Parasitaemia 11.19±0.38 11.19±0.38 9.83±0.19 8.63±0.17 6.52±0.39 5.36±0.43 % Cure 1.06 1.06 36.09 52.84 68.15 67.14

VC 100 % Parasitaemia 11.35±0.12 12.45±0.13 11.07±0.12 9.14±0.27 7.60±0.38 6.12±0.34 % Cure -0.35 7.71 28.02 50.05 62.87 62.48 200 % Parasitaemia 11.45±0.38 12.14±0.41 10.40±0.37 8.69±0.18 6.70±0.19 5.24±0.20 % Cure -1.24 10.01 32.38 52.51 67.27 67.87 400 % Parasitaemia 11.03±0.33 11.80±0.41 9.78±0.27 8.51±0.37 7.05±0.42 5.24±0.61 % Cure 2.48 12.53 36.41 53.50 65.56 67.87

FC 100 % Parasitaemia 11.52±0.32 13.00±0.42 10.75±0.34 8.37±0.23 6.51±0.55 5.20±0.52 % Cure -1.86 3.63 30.10 54.26 68.20 68.12 200 % Parasitaemia 11.29±0.36 12.16±0.26 10.46±0.23 9.38±0.16 7.67±0.43 4.60±0.41 % Cure 0.18 9.86 31.99 48.74 62.53 87.98 400 % Parasitaemia 11.27±0.34 11.83±0.34 10.30±0.45 8.28±0.16 6.35±0.17 4.49±0.33 % Cure 0.35 12.31 33.03 54.75 68.98 72.47

LL 100 % Parasitaemia 10.90±0.29 11.74±0.46 10.65±0.31 10.12±0.14 9.08±0.37 8.33±0.06 % Cure 3.63 12.97 30.75 44.70 55.64 48.93 200 % Parasitaemia 11.43±0.25 11.61±0.22 10.88±0.20 9.69±0.24 8.70±0.27 7.46±0.35 % Cure -1.06 16.16 29.26 47.05 57.50 54.26 400 % Parasitaemia 10.20±0.69 11.06±0.65 9.86±0.64 8.93±0.39 7.67±0.24 6.20±0.16 % Cure 9.80 18.01 35.89 51.20 62.53 61.99 Results are Mean count ± SEM; n = 5; p < 0.05

There is little or no scientific documentation of 1:1:1:1) to validate their ethnomedicinal claims. therapeutic benefits, safety information of such Comparative antimalarial studies of the various test combination or its advantage over monotherapy extracts using two in vivo models showed that the of the individual plants23. The number of medicinal individual plant extracts and co-combination to be plants that constitutes a polyherbal mixture or potential antimalarial agents. The two models combination could be dependent on patient employed were, 4-day suppressive test for assaying presentation or traditional health giver discretion24. candidates for early infection and Rane test which Nefang, is a polyherbal antimalarial product in evaluates curative effects of drug agents on South West Region of and consist of established infection26. 6 different antimalarial herbs25. The results indicated that all individual The present study evaluated anti-malarial administration of the extracts of the 4 plants and the properties of 4 Nigerian medicinal plants, Lophira combination of the 4 plants significantly (p < 0.05) lanceolate (LL), Protea madiensis (PM), Vernonia and dose-dependently inhibited growth of the parasite conferta (VC) and Fadogia cienkowskii (FC) (chemosuppression). Maximum growth inhibition individually and in combination (at fixed dose ratio of (93.44 %) of P. berghei was observed at 400 mg kg-1 ORABUEZE et al.: ANTIMALARIAL POTENTIAL OF NIGERIAN POLY HERBAL COMBINATION 721

Table 5 — Mean survival time (MST) of each group on day situation. However, PM, LL and VC, were sign of 30 (D0-D29) quick recrudescence was noticed 24 post last dose Drug Dose mg kg-1 Mean survival time (MST) administration may need a supporting drug for of each group prolonged and sustained antimalarial effect. The 5% DMSO 0.2 mL 7.63 ± 1.1 extract of FC showed an increased significant curative CQ 10 28.00 ± 0.0 effect 24 h post last dose administration, suggesting a FLVP 100 19.11 ± 1.37 long acting potential. The curative activity of FLVP 200 20.34 ± 0.56 indicated an early onset and prolonged chemo 400 24.88 ± 1.4 reduction of parasitaemia. Prolonged parasite FC 100 17.28 ± 1.63 reduction effect of FLVP may have been gotten from 200 17.52 ± 1.02 FC characteristics. This may explained the more 400 21.41 ± 1.02 prolonged MST recorded for FLVP than the LL 100 9.31 ± 1.36 individual monotherapies (Table 5). Prolonged MST 200 9.78 ± 0.87 of the test animals suggest that they lived longer 400 12.73 ± 2.24 compared with the negative control but were not fully VC 100 11.01 ± 1.34 protected by the administered drug/extract. The good 200 14.26 ± 1.40 curative potentials of FC and FLVP extracts suggest 400 15.78 ± 0.98 their ability to destroy trophozoites in the blood PM 100 12.73 ± 1.50 and/or prevent formation of gametocytes, thereby 27 200 13. 56 ± 1.21 preventing re-infection of the mosquitoes and man . 400 17.05 ± 2.01 Previous studies reported that individual medicinal Data are expressed as mean ± SEM plant components of polyherbal therapy could be 24 of Fadogia cienkowskii (FC) which is significantly acting as antimalarial, supporting agents or both . (p < 0.05) higher that the effect obtained of the Ethnobotanical claims of the 4 plants that constituted 2 standard drugs used CQ (89.36 %) and Art (85.46 %). FLVP indicated that some of the plants may be acting Chemosuppressive effects of FC and FLVP at both as antimalarial and supporting agents. 400 mg kg-1 gave 93.44 and 86.52 %, respectively. Antimicrobial, moderate antimalarial, antioxidants, antipyrexia, anemia and management of tiredness Though, the result demonstrated that FLVP produced 28,29,30 less effect compared to FC, however, a lesser amount activities of L. lanceolata have been reported . of the individual medicinal plants were required to These properties of LL could be useful in make up 400 mg kg-1 of FLVP. Thus, reducing the management of malaria fever complications and amount of each individual medicinal plant in the symptoms. Traditional and scientific documented uses formulation of FLVP could mean increase in safety, of FC extract as analgesic, blood building, tolerability and patient acceptability. The observed antipyrexia, relief of headache, management of control in weight loss in both FC and FLVP treated general body debility support it’s being included in 18,31 groups may be due to the ability of the test extracts to FLVP . A species, Fadogia agrestis leaves has inhibit the growth of parasitaemia after parasite been reported to exhibit in vitro antiplasmodial 28 inoculation, thereby protecting the animals from the activity against Plasmodium falciparum . Some negative effects of malaria. species of Fadogia have been implicated in gousiekte The co-administration of 4 medicinal plants poisoning studies but toxicity has been linked to (FLVP) in curative model significantly reduced the seasonal variation, few species, high moisture content 32 parasite load on daily bases and enhanced the survival and very large dose (one-third of body weight) . The time of the mice compared to those given the Vernonia has documented antimalarial and individual extracts in monotherapy. malarial symptom relief activities for some of its 16 Reduction of parasitaemia in single and species . Documented exploitation of therapeutic combination (FLVP) treated groups showed an early potentials of most of the plants used in this study is onset (24 h post initiation of drugs) of antiplasmodial limited and there is a need to explore more possible effect in established infection model. This indicates medical and otherwise usage. that no booster dose may be needed to initiate activity Similar records of combined extracts giving good and suggest that the drugs can be used in emergency clearance and suppression have been reported by 722 INDIAN J TRADIT KNOWLE, VOL. 17, NO. 4, OCTOBER 2018

several authors which include Kingsley et al., (2012), and Drug Development (Inter CEDD), Enugu state, Paula et al., (2012) and Osei-Djarbeng et al., Nigeria for their aid in collection and identification of (2015)33,8,23. No sign of toxicity or death was observed the plants. in oral acute toxicity evaluation of FLVP polyherbal combination given up to 4,000 mg kg-1. No literature References search indicated toxicity of any of the plants that 1 Walther B & Walther M, What does it take to control constituted FLVP poly herbal combination13,18. malaria?, Ann Trop Med Parasito, 101(8) (2007) 657–672. Many bioactive plant secondary metabolites are 2 Rowe AK, Steketee RW, Arnold F, Wardlaw T, Basu S, responsible for the observed anti-malarial activity but Bakyaita N, Lama M, Winston CA, Lynch M, Cibulskis RE, Shibuya K, Ratcliffe AA & Nahlen BL, Roll back malaria the most important and diverse bio-potencies have monitoring and evaluation reference group, viewpoint: been observed in alkaloids, quassinoids, flavonoids evaluating the impact of malaria control efforts on mortality and sesquiterpene lactones34. All the 4 plants that in sub-Saharan Africa, Trop Med Int Health, 12 (12) (2007) constituted FLVP presented alkaloid and flavonoid. 1524–1539 . Our study appears to be the first scientific 3 Vadalia J, Vekariya S, Raval M & Sheth N, In vitro cystein protease inhibitory activity of selected Indian antimalarial investigation report on antimalarial effects of Fadogia plants, Indian J Tradit Knowle, 16 (3) (2017) 476–481. cienkowskii and Protea madiensis, to the best of the 4 Murphy SC & Breman JG, Gaps in the childhood malaria authors knowledge. burden in Africa: adding cerebral malaria, neurological sequelae, anemia, respiratory distress, hypoglycemia and Conclusion complications of pregnancy to the calculus, Am J Trop Med Hyg, 64 (Suppl 1) (2001) 57–67. Although it is premature to conclude at this stage 5 WHO, World malaria report, World Health Organization, that this herbal combination can be used as effective Geneva, 2016. antimalarial, the findings provide a foundation 6 Wongsrichanalai C, Pickard AL, Wernsdorfer WH & for further exploration of possible therapeutic benefit Meshnick SR, Epidemiology of drug-resistant malaria, of herb-herb combinations for prevention and Lancet Infect Dis, 2 (2002) 209–218. management of malarial infections. The results 7 Thayer AM, Fighting malaria, Chem Eng News, 83 (2005) obtained in this study may not have indicated 69–82. synergistic or remarkable additive effect but showed 8 Bhagat M, Gupta S, Jamwal VS, Sharma S, Kattal M, Dawa S, Devi R & Bindu K, Comparative study on chemical that the medicinal plants that constituted FLVP may profiling and antimicrobial properties of essential oils from have contributed diverse pharmacological actions different parts of Eucalyptus lanceolatus, Indian J Tradit which may have contributed towards a more stable Knowle, 15 (3) (2016) 425–432 and long acting healing effect. Thus, the end product 9 Paula MC, William C, Paschal E & Peter S, In vitro may possibly slowed down the rate of development of antiplasmodial activities of extracts from five plants used singly and in combination against Plasmodium falciparum resistance, protect against side effects and increased parasites, J Med Plants, 6 (47) (2012) 770–5779. survival rate of the animal. This study therefore, 10 Adesegun SA, Orabueze IC & Coker HA, Antimalarial and offers a scientific basis for the traditional use of antioxidant potentials of extract and fractions of aerial part of these herbs separately and in combination against Borreria ocymoides DC (Rubiaceae), Pharmacogn J, 9 (4) malaria parasite. Further studies on the herbs, (2017) 534–540 safety, pharmacokinetic properties and optimizing its 11 Igboeli N, Onyeto CA, Okorie NA, Mbaoji FN, Nwabunike IA & Alagboso DI, Antidiarrheal activity of composition for maximum efficacy is highly methanol leaf extract of Lophira Lanceolata Tiegh recommended towards getting a better end product. (Ochnaeceae), Merit Res J Environ Sci Toxicol, 3 (4) (2015) 059–064. Conflict of Interests 12 Sani AA, Ibrahim MS, Mohammed I, Abdul KH, We declare that we have no conflict of interest. Ojuolape RA & Abdulkareem SS, Antimicrobial studies of aqueous extract of the leaves of Lophira lanceolata, Res J Pharm Biol Chem Sci, 2 (1) (2011) 637–643. Acknowledgement 13 Arbonnier M, Trees, and Lianas of West African dry The authors acknowledge and are grateful for the zones, Cirad, Margraf publishers GMBH, MNHN, financial support of TETFUND in the form of Wageningen, Netherlands, 2002, p. 436. research grant No: CRC/TETFUND/No. 2014/01. 14 Udegbunam SO, Okoli R, Chukwujekwu K, Anika SM, Udegbunam RI, Nnaji TO & Anyanwu MU, Evaluation of The authors are indebted to Mr. Ozioko F. and wound healing potential of methanolic Crinum jagus bulb Nwafor F. of International Centre for Ethnomedicine extract, J Intercult Ethnopharmacol, 4 (3) (2015) 194–201. ORABUEZE et al.: ANTIMALARIAL POTENTIAL OF NIGERIAN POLY HERBAL COMBINATION 723

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