EVALUATION OF IN-VITRO ANTIPLASMODIAL ACTIVITY OF SELECTED ETHNOBOTANICALLY IMPORTANT MEDICINAL PLANT EXTRACTS

EVALUATION OF IN-VITRO ANTIPLASMODIAL ACTIVITY OF SELECTED ETHNOBOTANICALLY IMPORTANT MEDICINAL PLANT EXTRACTS

Cheryl Sachdeva, Sunny Dhir, Amit Kaushik, Vineet Kaswan, Yashika Walia and Naveen Kumar Kaushik *

Amity Institute of Virology and Immunology, Amity University Uttar Pradesh, Sector-125, Noida, Uttar Pradesh, India.

ABSTRACT: Purpose: A serious cause of morbidity and mortality, malaria remains a major public health concern. Failure of available treatments due to the rapid emergence of drug-resistant parasite strains emphasizes the need for a continuous search for new, safe, accessible, and affordable drug treatments. Methods: In this context, an ethnobotanical survey was conducted, different parts of seventeen medicinal plants were collected, and their methanolic extracts were prepared. These extracts were evaluated for anti-plasmodial activity against two strains of Plasmodium falciparum: chloroquine-sensitive 3D7 and resistant INDO using SYBR-Green I assay. The extracts were also assessed for cytotoxic effects against HEK293 mammalian cell lines using MTT assay. Results: Two plant extracts: Albizia lebbeck and Tecomella undulata exhibited good activity with IC₅₀ ranging from 10-20µg/ml. Among others, eight showed moderate activity (IC_50­= 20.1- 50µg/ml), while eight extracts either showed poor activity or were not active upto IC₅₀=100µg/ml against Pf3 D7. The extracts with good to moderate effects showed equipotent effects against Pf INDO and were non-toxic to mammalian cell lines with selectivity indices ranging from 2.3 to >13.3. This study validated the traditional usage of the selected plants especially leaves of A. lebbeck and T. undulata. Conclusion: The results obtained have presented a starting material for identification and purification of active compounds that might provide an alternative drug therapy to fight against malaria.

Keywords: Antiplasmodial, Medicinal Plants, Albizia lebbeck, SYBR-Green I, Tecomella undulata, Cytotoxicity, Chloroquine resistance

INTRODUCTION: Malaria continues to be the most prevalent parasitic disease. Currently, 87 countries are malaria- affected with children below the age of five being most vulnerable as they accounted for 67% of deaths ¹.

Although recent advancements in the treatment and control strategies malaria remains a major concern due to the constant emergence of resistant Plasmodium strains against available antimalarials ^2-4.

The challenges of developing new therapeutics to reduce disease severity and eradicate malaria persist. Out of five Plasmodium species that cause human malaria, P. falciparum has developed resistance to almost all the available drugs, including current frontline artemisinin derivatives ⁵. This constant emergence of resistant strains and difficulty in developing an effective vaccine highlights the need for novel antimalarial compounds. Human life depends upon four basic things i.e., food, clothes, shelter and good health. The fourth one is provided by plant kingdom and that is why nature and natural products are foremost solution in healthcare system for centuries ⁶. Traditional knowledge and application of medicinal plants to treat diseases has increased success rate of drug development. Previous accomplishments of isolating quinine from bark of Cinchona tree and artemisinin from Artemisia annua are good examples of natural products as a source of lead candidates for drug development ^{7, 8}. Therefore, a plant product having specific clinical activity is considered as an initial point for a discovery and development.

India is among the richest countries in flora and since time immemorial, numerous medicinal plants have been used to treat a wide variety of diseases ^9-11. Therefore, the present study aimed to collect and screen medicinal plants from different regions of India against Plasmodium falciparum to identify a potent source for developing lead antimalarial candidates. A total of seventeen medicinal plants were tested against CQ- sensitive strain and CQ resistant strain of P. falciparum. The plants were also found to be non-cytotoxic to mammalian cells. The results obtained in this study reveal medicinal plants in India that have antimalarial potential. On further evaluation, they would lead to the identification and isolation of novel, a cost-effective antimalarial drug.

MATERIALS AND METHODS:

Collection of Medicinal Plants: An ethnobotanical survey using standardized questionnaire ¹¹ was conducted at Kasauli, Himachal Pradesh; Noida, Uttar Pradesh; Sanjay Van, New Delhi. Locals were interviewed to identify plants that are traditionally used for treating malaria-associated symptoms such as fever, chills, anemia. Following standard sustainable collection protocol, plant materials were collected and identified by Dr. Sunny Dhir, Maharishi Markandeshwar (Deemed to be University), Mullana, India using reference samples that were preserved and deposited at Amity Institute of Virology and Immunology, Amity University Uttar Pradesh, Noida, Uttar Pradesh, India.

Preparation of Medicinal Plant Extracts: Each plant part was air dried in shade and powdered in a grinder. Dried and powdered plant material were extracted using methanol as solvent of extraction ^{11, 12}. Briefly, powdered leaf samples (12- 16g) were suspended in 200 ml of methanol and kept for overnight shaking in orbital shaker. The filtrate was then collected using Whatman filter paper No. 1 and the residue was again suspended in 200 ml methanol the process was repeated 5 times to extract each sample in total 1L(200 ml × 5) of methanol. The filtrate collected from each sample was pooled and, solvent from the filtrate was then evaporated using rotary-evaporator at 42°C. The obtained dry residue was weighed and stored at 4°C.

In-vitro P. falciparum Culture Maintenance: Two strains of Plasmodium falciparum namely3D7 (CQ sensitive) and INDO (CQ resistant) were bought from Malaria Research and Reference Reagent Resource Center (MR4) and cultivated in laboratory set up using a modified Trager and Jenson method ¹³. The parasites were suspended in fresh erythrocytes (obtained from Rotary Blood Bank, Tughlakabad Institutional Area, New Delhi, India) at 4% hematocrit using complete media. Complete media was prepared by combining 16.2 g/L Roswell Park Memorial Institute (RPMI) 1640 powder (Gibco), 0.5% albumax I (Gibco), 0.2% sodium bicarbonate, 0.005% hypoxanthine (MERCK), 10mg/l gentamicin. The culture was then incubated at 37°C and a gaseous mixture of 5% O₂, 5% CO₂ and 95% N₂. Media was refreshed daily and parasitemia was monitored microscopically using Giemsa stain.

Preparation of Test Samples: Stock solutions of test samples (25 mg/ml) were prepared in dimethyl sulfoxide (DMSO). Ten-fold dilution from the stocks were prepared to obtain 2.5 mg/ml in 10% DMSO. Following this, two-fold dilutions were prepared in a culture medium and 4 µl from each of these were added to the assay plated to obtain working concentrations of 100 -12.5 µg/ml. The final concentration of DMSO in each well was 0.4% which is found to be non-toxic to the parasite. For controls: stocks of artemisinin were prepared in DMSO and that of CQ in sterilized distilled water.

Growth Inhibition Assay against P. falciparum: Antimalarial efficiency of methanolic extracts was tested against P. falciparum using SYBR green I fluorescence-based assay ¹⁴. Using 5% sorbitol, parasites were synchronized at ring stages and incubated at 2% hematocrit, 1% parasitemiain a 96-well plate. Dilutions of each of the test samples were then added to this in triplicates in increasing order of concentration. DMSO (0.4%) was used as a negative control. For positive controls: 50nM artemisinin and 100nM CQ were used. Following this, the test plates were incubated at 37°C with standard culture conditions for 48 h. After incubation, 100 µl SYBR Green I solution was added to each well following which the plates were incubated at 37°C in dark for 1 h. Subsequently, fluorescence was measured on Perkin Elmer fluorescence plate reader at 485 nm excitation and at 530 nm emission. Dose-response curves were plotted using fluorescence counts and drug concentration to determine the inhibitory concentration (IC₅₀).

In-vitro Cytotoxicity of the Plant Extracts: Cytotoxic effects of plant extracts were evaluated on HEK293 mammalian cell lines using MTT assay ¹⁵. The cell lines were maintained in complete media containing DMEM (Gibco), 10% fetal bovine serum (Gibco), 100 Units/ml penicillin, 100 µg/ml streptomycin (Gibco) and incubated at 37 C, 5% CO₂. For the assay, cells were trypsinized using 0.25% trypsin (Gibco), and 10⁴ cells/100µl/well were seeded in a 96-well tissue culture-treated flat bottom plate. The plates were incubated at standard culture conditions. Following 24 h incubation, test samples in the concentration of 200 µg/ml to 25 µg/l, in triplicates, were added to each well, and plates were incubated for 24 h. Subsequently, 0.5 mg/ml of MTT-PBS solution was added to the wells and the plates were then incubated for 4 h. After this, 100 µl of DMSO was added to every single well and mixed thoroughly to solubilize formazan crystals. The absorbance was read at 570nm using a multi-well plate reader, and dose-response curves were plotted to determine TC₅₀.

RESULTS:

Survey and Collection of Samples: After conducting an ethnobotanical survey at three different locations in India and interviewing 50 locals, details of plants used to relieve headache, fever, nausea, etc. were recorded. In addition, details of methods used to prepare the extracts, mode of administration, doses administered were also collected. Based on that, eighteen parts from seventeen plantswere collected to test their antimalarial potential Fig. 1, Table 1.

FIG. 1: SITES OF PLANT SAMPLE COLLECTION

TABLE 1: TRADITIONAL USES OF SELECTED MEDICINAL PLANTS

In-vitro Antiplasmodial and Cytotoxic Effects of the Extracts: Out of eighteen plant extracts assessed against Pf3D7, leaves of two plants Tecomella undulata and Albizia lebbeck showed good activities with IC₅₀ ranging from10.1 to 20µg/ml, eight extracts namely, leaves of Nerium oleander, Ricinus communis, Saccharum officinarum, Solanum lycopersicum, Citrus pseudolimonum, Bambusa vulgaris, Urtica dioica, Grewia optiva were moderately active with IC₅₀= 20.1 to 50 µg/ml, four extracts viz., leaves of Ficus benjamina, Catharanthus roseus, Spinacia oleracea, Butea monosperma showed poor antimalarial effects with IC₅₀ ranging from 50-100µg/ml and four plant extracts namely, leaves of Triticum aestivum, Anethum graveolens and leaf and flower extracts of Gaillardia aristata were not active up to 100µg/ml Table 2, Fig. 2.

FIG. 2: DOSE-DEPENDENT GROWTH INHIBITION CURVES OF PLASMODIUM FALCIPARUM TREATED WITH PLANT EXTRACTS. CURVES OF PF3D7 TREATED WITH METHANOLIC  EXTRACTS  OF (A) LEAVES OF A. LEBBECK [AL(L)], B. VULGARIS [BV(L)], B. MONOSPERMA[BM(L)], C. ROSEUS [CR(L)], C. PSEUDOLIMONUM [CP(L)], F. BENJAMINA FB(L)]; (B) LEAVES AND FLOWER OF G. ARISTATA [GA(L) AND GA(F)], LEAVES OF A. GRAVEOLENS G. OPTIVA[GO(L)], N. OLEANDER [NO(L)], R. COMMUNIS [RC(L)]; (C) LEAVES OF S. OFFICINARUM [SOF(L)], S. OLERACEAE [SOL(L)], S. LYCOPERSICUM [SL(L)], T. UNDULATE [TU(L)], T. AESTIVUM [TV(L)], U. DIOICA [UD(L)]

Ten extracts that exhibited good to moderate effects against Pf3D7 were assessed against CQ resistant PfINDO and six of them i.e., T. undulata, A. lebbeck, N. oleander, C. pseudolimonum, G. optiva and B. vulgaris showed were found to be equipotent with IC₅₀< 50 µg/ml. These extracts were also evaluated against HEK293 mammalian cell lines and were found to be non-toxic with selectivity index ranging from 2.3 to >13.3 Table 2.

TABLE 2: IN-VITRO ANTIPLASMODIAL AND CYTOTOXIC ACTIVITY OF SELECTED MEDICINAL PLANTS

* Numbers in parenthesis denote resistance index (IC₅₀PfINDO/IC₅₀Pf3D7), # Numbers in parenthesis denote SI (TC₅₀ HEK293/IC₅₀Pf3D7).

DISCUSSION: Natural products have always played a important part in disease management as plant-derived metabolites are used for treating wide range of diseases ^{9, 16}. Hence, drug discovery begins with collection of plants on the basis on their traditional knowledge. In this context, seventeen medicinal plants were screened for antimalarial activity against CQ-sensitive 3D7 and CQ resistant INDO strain of Plasmodium falciparum. Albizia lebbeck (L.) Benth. (Mimosoideae) is a traditional medicine for arthritis, asthma, cold and cough, cancer ^{17, 18}. Here, A. lebbeck showed good activity with IC₅₀ = 17 and 18.1 µg/ml against both Pf3D7 and Pf INDO respectively. Active antimalarial nature of A.lebbeckhas previously been recognized by Kalia et. al who noted significant in-vitro activity with IC₅₀= 8.2 and 5.1 µg/ml against PfMRC2 (CQ sensitive) and PfRKL9 (CQ resistant) strain of P. falciparum by ethanolic extract of this plant ¹⁹.

Tecomella undulata (Sm.) Seem (Bignoniaceae) is a well-recognized medicinal plant traditionally used to treat liver disorders, cancer, hemorrhoids, diabetes, as a blood purifier ²⁰. In our study, T. undulata showed good antiplasmodial activity against both Pf3D7 and PfINDO with IC₅₀= 15 µg/ml and 15.3 µg/ml respectively.

Saccharum officinarum is a constituent of polyherbal formulation known as SAABMAL used as an antimalarial in Nigeria. Recently, a study showed 95% suppression of parasitemia on treating P. berghei-infected mice with 400mg/kg of SAABMAL ²¹. Our study, however, showed moderate activity of leaves of S. officinarum with IC₅₀= 50 and 56 μg/ml against Pf3D7 and PfINDO respectively.

Leaf water extract of F. benjamina collected from Cameroon showed significant antimalarial effects with IC₅₀= 12 μg/ml and 26 μg/ml against Pf3D7 and PfINDO; petroleum ether extract of leaves collected from Punjab, India showed IC₅₀=14 μg/ml which improved substantially on bio-guided fractionation with IC₅₀=4 μg/ml of hexane fraction and 7 μg/ml of chloroform fraction against Pf3D7 ²³. On the contrary, our study, where both location and solvent used for extraction were different, showed poor activity of leaf methanolic extract with IC₅₀=100 μg/ml against Pf3D7.

Nerium oleander L. is a drought-tolerant plant and is known to have numerous bioactivities such as anti-inflammatory, antibacterial, immuno-modulatory, neuroprotective, etc. Here, we have shown good to moderate antiplasmodial effects of leaves of N. oleander. Previous studies have reported the active nature of this plant against malaria vectors, i.e., larval stages of Anopheles stephensi with LC₅₀- 0.58 g/l of chloroform leaf extract, 0.55 g/l of benzene flower extract ²⁴, 94.6 mg/l of acetone flower extract ²⁵ and larval stages of A. gambiae with LC₅₀= 127.8 ppm of aqueous leaf extract and 281.5 ppm of aqueous flower extract ²⁶.

This plant is extremely poisonous due to the presence of toxic compounds such as oleandrin ^{27, 28}; however, leaf methanolic extract in our study was found to be non-toxic to mammalian cell lines and moderately toxic against malaria parasite. Likewise, Ricinus communis, which is widely distributed throughout tropics and temperate regions of the world has shown to be effective against malaria vector. Sogan et al, observed 35 and 45% mortality of A. culicifacies on treatment with leaf (LC₅₀= 65.62 ppm) and seed (LC_50­= 9.37 ppm) dichloromethane extract of R. communis respectively. Further, treatment with aqueous seed extract on Day 1 showed 65- 100% mortality of A. gambiae for 5 days with LD₅₀= 74.1 ppm ²⁶. However, antiplasmodial of R. communis has not been evaluated yet, and in the present study, we showed moderate effects of leaf extract against Pf3D7and good effects against PfINDO.

Grewia species are widely distributed in tropical and subtropical regions and are among the most important ingredients in traditional medicines. Many species of Grewia have been reported to exhibit antimalarial activity for instance chloroform fraction of Grewiabila mellata collected by chromatography inhibited growth of D6 and W2 strains of P. falciparum with IC₅₀=2.3 and 1.7 µM respectively ²⁹. However, G. optiva has not been studied yet despite its common use as a traditional herb to treat malaria. In our study, methanolic crude leaf extract showed moderate activity which might be attributed to its constituents. Solanum lycopersicum originated in Andes and now is common all around the world. This economically important crop has a high nutritive value and has been found to exhibit bioactivities such as antibacterial, antioxidant ³⁰. Larvicidal activity of S. lycopersicum has been reported against Aedesand Culex ³¹; however, antimalarial or larvicidal against malaria vector has not been assessed yet. Here, leaves of this plant has shown moderate effects against Plasmodium parasite.

Urtica dioica is a widespread perennial plant that grows on light soil and humus. Extracts of U. dioica have been used for treating anemia, pain, rheumatism ³². It has also exhibited antiviral properties against dengue virus ³³. Despite its traditional usage, antimalarial potential of this has not been evaluated yet. Here, leaf extract showed moderate effects against P. falciparum validating its traditional usage.

Bambusa vulgaris is the widely distributed throughout the tropical and subtropical regions and is the most common among all Bambusa species. The leaves of this plant are traditionally used to treat malaria ³⁴, typhoid ³⁵, venereal diseases ³⁶, haematuria as a febrifuge agent. Previous studies have shown that aqueous leaf extracts from Ghana and ethanolic leaf extract from Cuba inhibited growth of Pf3D7 with IC₅₀= 7.5 and 4.7 µg/ml, respectively ^{37, 38}. Additionally, Anigboro et al; reported a decrease in parasite load by 87% compared to positive control on treatment with 300 mg/kg of ethanolic leaf extract of B. vulgaris ³⁹. However, in our study, methanolic leaf extracts exhibited moderate antimalarial effects with IC₅₀= 30 µg/ml. The variation in activity might be attributed to: 1) choice of solvents that extracted out different components and their interaction influenced their activity, 2) different locations of sample collection.

Butea monosperma is a deciduous tree widely distributed in India, Ceylon, Burma. This plant is used in Unani, Ayurveda, Homeopathy and modern medicine ⁴⁰. The plant is used to treat helminthiasis, filariasis, diarrhea, and dysentery ⁴¹. Previous studies have reported in vivo activity of this plant where a methanolic extract of leaves of B. monosperma showed only 6% suppression of P. berghei parasitemia on day 4 at a dosage of 500 mg/kg ⁴². In line with that, in our study, the leaf extract of B. monosperma showed poor in-vitro activity against Pf3D7.

CONCLUSION: Rampant multi-drug resistant Plasmodium parasites calls for the continuous search of novel antimalarial drugs. This study revealed remarkable antimalarial efficacy of selected plants that were used by traditional healers and studied. In-vitro assessment of two medicinal plants: A. lebbeck and T. undulata showed antimalarial potential that justify their usage and healing properties against malaria-like symptoms. The antimalarial activity of these plants might be due to the molecules present in the plant extracts. Therefore, further studies on isolation and characterization of the principal components contributing to their active nature are likely to yield novel compounds that could be developed as alternative chemotherapy for treating malaria.

Statements & Declarations:

Funding: This work was supported by the Science and Engineering Research Board, Department of science and technology, Government of India. Grant numbers YSS/2015/000728

Competing Interests: The authors have no relevant financial or non-financial interests to disclose.

ACKNOWLEDGEMENT: The authors thank the Science & Engineering Research Board-Department of Science and Technology (SERB-DST), Government of India, for funding the study. The authors would also like to thank Ms. Ruhi Thakur, Punjabi University, for helping with the sample collection.

CONFLICTS OF INTERESTS: Authors Declares No conflicts of interest.

REFERENCES:

1. WHO: World Malaria Report 2020. https://www.who.int/publications/i/item/9789240015791
2. White NJ, Pukrittayakamee S, Hien TT, Faiz MA, Mokuolu OA and Dondorp AM: Malaria. Lancet. 2014; 383(9918): 723-735. doi:10.1016/S0140-6736(13)60024-0
3. Menard D and Dondorp A: Antimalarial drug resistance: a threat to malaria elimination. Cold Spring Harbor Perspectives in Medicine 2017; 7(7): 025619.
4. Tang YQ, Ye Q, Huang H and Zheng WY: An overview of available antimalarials: discovery, mode of action and drug resistance. Current Molecular Medicine. 2020; 20(8): 583-92.
5. Duffey M, Blasco B, Burrows JN, Wells TN, Fidock DA and Leroy D: Assessing risks of Plasmodium falciparum resistance to select next-generation antimalarials. Trends in Parasitology 2021; 37(8): 709-21.
6. Sachdeva C and Kaushik NK: Spices-Reservoir of Health Benefits. Natural Medicinal Plants 2022; 11: 255.
7. Randrianarivelojosia M, Raveloson A, Randriamanantena A, Juliano JJ, Andrianjafy T, Raharimalala LA and Robert V: Lessons learnt from the six decades of chloroquine use (1945–2005) to control malaria in Madagascar. Transactions of the Royal Society of Tropical Medicine and Hygiene 2009; 103(1): 3-10.
8. Eastman RT and Fidock DA: Artemisinin-based combination therapies: a vital tool in efforts to eliminate malaria. Nature Reviews Microbiology 2009; 7(12): 864-74.
9. Pitchai D, Manikkam R, Rajendran SR and Pitchai G: Database on pharmacophore analysis of active principles, from medicinal plants. Bioinformation 2010; 5(2): 43.
10. Sofowora A, Ogunbodede E and Onayade A: The role and place of medicinal plants in the strategies for disease prevention. African Journal of Traditional, Complementary and Alternative Medicines 2013; 10(5): 210-29.
11. Sachdeva C, Mohanakrishnan D, Kumar S and Kaushik NK: Assessment of in-vitro and in-vivo antimalarial efficacy and GC-fingerprints of selected medicinal plant extracts. Experimental Parasitology 2020; 219: 108011.
12. Sachdeva C, Kumar Sand Kaushik NK: Exploration of anti-plasmodial activity of Prunus cerasoides-Ham. ex D. Don (family: Rosaceae) and its wood chromatographic fractions. Acta Parasitologica. 2021; 66(1): 205-12.
13. Trager W and Jenson JB: Cultivation of malarial parasites. Nature 1978; 273(5664): 621-2.
14. Smilkstein M, Sriwilaijaroen N, Kelly JX, Wilairat P and Riscoe M: Simple and inexpensive fluorescence-based technique for high-throughput antimalarial drug screening. Antimicrobial Agents and Chemotherapy 2004; 48(5): 1803-6.
15. Mosdam TJ: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxic assay. J. Immunol. Methods 1983; 65: 55-63.
16. Drewry DH and Macarron R: Enhancements of screening collections to address areas of unmet medical need: an industry perspective. Current Opinion in Chemical Biology 2010; 14(3): 289-98.
17. Desai TH and Joshi SV: Anticancer activity of saponin isolated from Albizialebbeck using various in-vitro Journal of Ethnopharmacology 2019; 231: 494-502.
18. Bobby MN, Wesely E and Johnson M: High performance thin layer chromatography profile studies on the alkaloids of Albizialebbeck. Asian Pacific Journal of Tropical Biomedicine 2012; 2: 1-6.
19. Kalia S, Walter NS and Bagai U: Antimalarial efficacy of Albizialebbeck (Leguminosae) against Plasmodium falciparum in-vitro & P. berghei in-vivo. The Indian Journal of Medical Research 2015; 142(1): 101.
20. Jain M, Kapadia R, Jadeja RN, Thounaojam MC, Devkar RV and Mishra SH: Traditional uses, phytochemistry and pharmacology of Tecomella undulata–A review. Asian Pacific Journal of Tropical Biomedicine 2012; 2(3): 1918-23.
21. Obidike IC, Amodu B and Emeje MO: Antimalarial properties of SAABMAL®: An ethnomedicinal polyherbal formulation for the treatment of uncomplicated malaria infection in the tropics. The Indian Journal of Medical Research 2015; 141(2): 221.
22. Rufin Marie TK, Mbetyoumoun Mfouapon H, Madiesse Kemgne EA, Jiatsa Mbouna CD, Tsouh Fokou PV, Sahal D and Fekam Boyom F: Anti-plasmodium falciparum activity of extracts from 10 Cameroonian medicinal plants. Medicines 2018; 5(4): 115.
23. Mukhtar HM, Singh A and Kaur H: Bioassay Guided Fractionation and in-vitro Anti-plasmodial Activity of Ficus deltoidea and Ficus benjamina. Pharmacognosy Journal 2018; 10(2).
24. Fakoorziba MR, Moemenbellah-Fard MD, Azizi K and Mokhtari F: Mosquitocidal efficacy of medicinal plant, Nerium oleander (Apocynaceae), leaf and flower extracts against malaria vector, Anopheles stephensi Liston (Diptera: Culicidae) larvae. Asian Pacific Journal of Tropical Disease 2015; 5(1): 33-7.
25. Raveen R, Pandeeswari M, Ahmed F, Reegan D, Tennyson S, Arivoli S and Jayakumar M: Bioefficacy of Nerium oleander Linnaeus (Apocynaceae) floral extracts on the larva of three vector mosquitoes of medical importance. International Journal of Mosquito Research 2017; 4(6): 65-77.
26. Kehail MA, Bashir NH, Abdelrahman EE and Abdelrahim AM: Larvicidal activity of three plants powders and aqueous extracts on Anopheles and Culex mosquito larvae (Diptera: Culicidae).International Journal of Mosquito Research 2017; 4(4): 37-41
27. Soto-Blanco B, Fontenele-Neto JD, Silva DM, Reis PF and Nóbrega JE: Acute cattle intoxication from Nerium oleander pods. Tropical Animal Health and Production 2006; 38(6): 451-4.
28. Barbosa RR, Fontenele-Neto JD and Soto-Blanco B: Toxicity in goats caused by oleander (Nerium oleander). Research in Veterinary Science. 2008; 85(2): 279-81.
29. Ma C, Zhang HJ, Tan GT, Hung NV, Cuong NM, Soejarto DD and Fong HH: Antimalarial compounds from Grewia bilamellata. Journal of Natural Products 2006; 69(3): 346-50.
30. Sajet AL-Oqaili RM and Salman BB: In-vitro antibacterial activity of Solanum lycopersicum extract against some pathogenic bacteria. In-vitro 2014; 27.
31. Maragatham M and Joseph D: Larvicidal evaluation of aged and fresh leaf extract of Solanum lycopersicum International Journal of Mosquito Research. 2019; 6: 57-60.
32. Ghaima KK, Hashim NM and Ali SA: Antibacterial and antioxidant activities of ethyl acetate extract of nettle (Urtica dioica) and dandelion (Taraxacum officinale). Journal of Applied Pharmaceutical Science 2013; 3(5): 096-9.
33. Flores-Ocelotl MR, Rosas-Murrieta NH, Moreno DA, Vallejo-Ruiz V, Reyes-Leyva J, Domínguez F and Santos-López G: Taraxacumofficinale and Urticadioica extracts inhibit dengue virus serotype 2 replication in-vitro. BMC Complementary and Alternative Medicine 2018; 18(1): 1-0.
34. Asase A and Asafo-Agyei T: Plants used for treatment of malaria in communities around the Bobiri forest reserve in Ghana. Journal of Herbs, Spices & Medicinal Plants 2011; 17(2): 85-106.
35. Ajaiyeoba EO, Oladepo O, Fawole OI, Bolaji OM, Akinboye DO, Ogundahunsi OA, Falade CO, Gbotosho GO, Itiola OA, Happi TC and Ebong OO: Cultural categorization of febrile illnesses in correlation with herbal remedies used for treatment in Southwestern Nigeria. Journal of Ethnopharmacology 2003; 85(2-3): 179-85.
36. Brink, M: Bambusa vulgaris ex J. C. Wendl. [Internet] Record from PROTA4U. Louppe, D., Oteng-Amoako, A.A. & Brink, M. (Editors). PROTA (Plant Resources of Tropical Africa / Ressourcesvégétales de l’Afriquetropicale), Wageningen, Netherlands. 2008 <http://www.prota4u.org/search.asp>. Accessed 22 June 2022.
37. Komlaga G, Cojean S, Dickson RA, Beniddir MA, Suyyagh-Albouz S, Mensah ML, Agyare C, Champy P and Loiseau PM: Antiplasmodial activity of selected medicinal plants used to treat malaria in Ghana. Parasitology Research 2016; 115(8): 3185-95. doi: 10.1007/s00436-016-5080-8.
38. Valdés AF, Martínez JM, Lizama RS, Gaitén YG, Rodríguez DA and Payrol JA: In-vitro antimalarial activity and cytotoxicity of some selected Cuban medicinal plants. Revista do Instituto de Medicina Tropical de Sao Paulo 2010; 52: 197-201.
39. Anigboro AA: Antimalarial efficacy and Chemopreventive capacity of Bamboo leaf (Bambusa vulgaris) in malaria parasitized mice. Journal of Applied Sciences and Environmental Management 2018; 22(7): 1141-5.
40. Sindhia VR and Bairwa R: Plant review: Butea monosperma. International Journal of Pharmaceutical and Clinical Research 2010; 2(2): 90-4.
41. Bhalla V and Walter H. Research bulletin of the Punjab university. Science 1999; 48: 87-94.
42. Samy K and Kadarkari M. Antimalarial activity of traditionally used Western Ghats plants from India and their interactions with chloroquine against chloroquine-tolerant Plasmodium berghei. Vector-Borne and Zoonotic Diseases 2011; 11(3): 259-68.
    

    ---

    How to cite this article:

    Sachdeva C, Dhir S, Kaushik A, Kaswan V, Walia Y and Kaushik NK: Evaluation of in-vitro antiplasmodial activity of selected ethnobotanically important medicinal plant extracts. Int J Pharm Sci & Res 2023; 14(2): 988-96. doi: 10.13040/IJPSR.0975-8232.14(2).988-96.

    ---

    All © 2023 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.