Antimalarial Effects of Ethanolic Leaf Extracts of Azadirachta indica and Ocimum gratissimum, and Their Histologic Effects on Some Organs (Liver, Kidney and Heart) of Plasmodium berghei Infected Albino Mice
Annual Research & Review in Biology,
Page 47-63
DOI:
10.9734/arrb/2023/v38i830602
Abstract
Azadirachta indica (Neem) and Ociumum gratissimum (clove Basil) have long been employed locally for the management of malaria. The present study compared antimalaria activities of the ethanolic leaf extracts of the individual plants, and assessed their combined effects on some organs of malaria-infected mice, at the Parasitology and Histopathology units, Federal Medical Centre (FMC), Owerri, from January to March, 2021. The leaves of the different plants were extracted with absolute ethanol (BDL 95%) for the test. Clean albino mice were experimentally infected intraperitoneally with chloroquine-sensitive Plasmodium berghei NK65 strain. Parasitaemia level was determined before parasite inoculation and at 24 hours post treatment period. Histopathological study on the liver, kidney, and heart was carried out using the Paraffin Setions method. Extracts of the leaves were administered orally, while chloroquine administration was intramuscular. The efficacy of the leaf extracts was tested on the P. berghei infected albino mice using the 4-day curative test. The lethal median dose (LD50) recorded for neem and clove basil leaf extracts were 31.62 and 1246.9 mg/kg body weight, respectively. Significant activity against the parasite was produced by infected mice treated with extracts of A. indica and O. gratissimum, and their combinations throughout the treatment period (P<.05). Highest reduction of parasitaemia was observed on day 4. Maximum parasitaemia reduction (78.65%) was attained with 30mg/kg of the combined extracts on the 7th day. Mild pathological lesions were observed in mice treated with A. indica leaf extract. These observations indicate better anti-malaria activity of the combination therapy as compared with the individual extracts of A. indica and O. grassimum, and indicate good antimalarial and protective roles of the plant extracts on the parasitized mice at large, as it slows down development of resistance.
Keywords:
- Malaria treatment
- phytotherapy
- drug resistance
- synergism of phytochemicals
- histology
- organs
How to Cite
C. C., A., A. E., O., E. O., O., & A. E., A. (2023). Antimalarial Effects of Ethanolic Leaf Extracts of Azadirachta indica and Ocimum gratissimum, and Their Histologic Effects on Some Organs (Liver, Kidney and Heart) of Plasmodium berghei Infected Albino Mice. Annual Research & Review in Biology, 38(8), 47-63. https://doi.org/10.9734/arrb/2023/v38i830602
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Adeyemo-Salami OA, Farambi EO, Ademowo OG. An investigation into the antimalarial effect of methanolic extract of Paullinaipinnata leaves in P. berghei infected mice and course of infection. African Journal of Medicine and Medical Sciences. 2014;43(1):93-94.
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Iwuchukwu IC, Vincent CN. Studies on prevalence of malaria and its adverse foetal outcomes in FMC, Owerri, Imo State, Nigeria.Archives of Community Medicine and Public Health. 2021; 7(2): 151-163.
Osonwa EU, Mbonu DO, Eluu CS, Oli AN. Antiplasmodial and biochemical effects of combination of ethanolic leave-extract of Azadirachta indicaand Ocimum gratissimum on Plasmodium berghei-infected mice. African Journal of Pharmaceutical Sciences and Pharmacy. 2017;5:1-6.
Afolabi OJ, Simon-oke IA, Oladokun OI. Antiplasmodial Activity of Ethanolic Extract of Neem Leaf (Azadirachta indica) in Albino Mice Infected with Plasmodium berghei. International Archives of Clinical Pharmacology. 2021;7:2572-2578.
Nwankwo EN, Ezekwesili OO, Chude CM. In vitro antiplasmodial and in vivo toxicity potentials of Mentha piperita and Ocimum gratissimum essential oils and their synergistic effect with conventional antimalarial drugs against Plasmodium falciparum. International Journal of Mosquito Research. 2022;9(1):114-122.
Omagha R, Idowu ET, Adeneye AK. Survey of ethnobotanical cocktails commonly used in the treatment of malaria in southwestern Nigeria. Future Journal of Pharmaceutical Sciences. 2021;7:152-158.
Nigussie G, Wale M. Medicinal plants used in traditional treatment of malaria in Ethiopia: a review of ethnomedicine, anti-malarial and toxicity studies. Malaria Journal. 2022;21:262-266.
Million E, Mulugeta T, Umeta B. Traditional medicine practice and its role in the management of malaria in Jimma Town, Oromia, Ethiopia. Journal of Infection and Drug Resistance. 2022;15:2187-2198.
Williamson, EM. Synergy and other interactions in Phytomedicines. Phytomedicine. 2001;8(5): 401-409.
Erhirhie, EO, Ikegbune, C, Okonkwo, OB. Antimalarial herbal drugs: a review of their interactions with conventional antimalarial drugs. International Journal of Phytomedicine and Physiotherapy. 2021;7:4-10.
Achi NK, Onyeabo C, Nnate DA, Ekeleme-Egedigwe CA, Kalu IK, Chibundu IC, Wokoma GC. Therapeutic effects of Azadirachta indica A. juss. leaves in malaria-induced male wister rats. Journal of Pharmacy and Pharmacognosy Research. 2018;6(3):191-204.
Mbugi EV, Sife AS, Mboni R, Grace EPM, Bestina D, Edda TL. Effectiveness of Azadirachta indica (neem tree) on prevention and treatment of clinical human malaria: A systematic review. Journal of East Africa Science. 2021;3:1-4.
Yarmohammadi F, Mehri S, Najan N, Salar AS, Hosseinzadeh, H. The protective effect of Azadirachta indica (Neem) against Metabolic Syndrome: A Review. Iran Journal of Basic Medical Sciences. 2021;24 (3):280-292.
Annette H, Barbara P, Sofia T, Judith N, Fabrizio, B. Effects of Azadirachta indica seed kernel extracts on early erythrocytic schizogony of Plasmodium berghei and proinflammatory response in inbred mice. Malaria Journal. 2019;18:35-43.
Alex DWA, Okorie PU. Water quality studies of Nworie Rivers in Owerri, Nigeria. Journal of Mississipi Academy of Sciences. 2008;20(1):55-60.
Ejebe DE, Emudianohwo JO, Ozoko T, Simialayi IM, Esume CO, Maduodi UV. An Investigation into the antiplasmodial effect of the ethanol extract of the leaves of Helliantusannus in Swiss Albino mice. Global Journal of Pharmacology. 2011;5 (2):92-96.
Peter IT, Anatoli VK. The current global malarial situation.Malaria parasite biology, pathogenesis, and protection. American Society for Microbiology, Washington DC. 1998;11-22.
Dawit D, Eyassu M, Asfaw D, Dawit A, Kelbessa U, Wallelign M, Daniel M, Ashenafi, A, yared, M. In vivo antimalaria activity of hydrocoholic extracts from Asparagus africanus Lam. In mice infected with Plasmodium berghei. Ethiopian Journal of Health Development. 2006; 20(2):112-118.
Ejebe DE, Siminialayi IM, Emudainowho JOT, omakporaye SL, Ojeih AE, Akonoghrere R, Odekuma IE, Ahatty GC. An improved technique for the oral administration of solutions of test substances to experimental rats using mediflon.medicut intravenous cannula. American Journal of Biotechnology. 2009; 8(6):60-96.
Lorke D. A New Approach to Practical Acute Toxicity Testing. Achives of Toxicology. 1983;54:275-287.
Matsumura, F. Toxicology of insecticides. Plenum press, York and London. 1975; 24.
Corbett JR, Wright, K, Baillie, AC. The Biochemical mode of Action of pesticides, 2nd, Academic press, London and New York; 1984.
David AF, Lip JR, Simon LC, Rato B, Solomon N. Antimalarial drug discovery: efficacy models for compound screening. Nature review. 2004;16:522-528.
Awe SO, Makinde, JM. Effect of petroleum ether fractions of Morindalucida on Plasmodium bergheiberghei in mice. Journal of Pharmaceutical Biology. 1998;36(4):301-304.
Tona L, Mesia, K, Ngimbi NP, Chrimwami,B, Okond A, Cimanga K, Bruyne, TD, Apers, S, Hermans, N, Totte, J, Pieters, L. In-vivo antimalarial activity of Cassia occidentalis, Morinda morindoide sand Phyllanthusniruri. Annals of Tropical Medicine and Parasitology. 2001;95(1) :47-57.
Iwalokun BA. Enhanced antimalarial effects of chloroquine by aqueous Vernonia amygdalina leaf extract in mice infected with chloroquine resistant and sensitive Plasmodium berghei strains. Journal of African Health Sciences. 2008; 8(1):25-35.
Adebajo, AC, Odediran, SA, Aliyu, AF, Nwafo, PA, Nwoko, TN, Umana, SU. In Vivo Antiplasmodium Potentials of the Combinations of four Nigerian Antimalarial Plants. Journal of Molecules. 2014;19(9): 13136-13146.
Somsak V, Chachiyo S, Kittitorn J, Audomkasok S, Sriwiphat S. Antimalarial and Antihypoglycemic Properties of Siamese Neem Tree (Azadirachta indica) in Plasmodium berghei-infected Mice. Malaria Contraction Elimination. 2015;4: 134-137.
Ekpo DE, Ekanemesang MU. Antiplasmodial/antimarlarial effect of ethanol extracts of leaves of Vernonia amygdalina and Gongronema latifolium on the activity of catalase in Plasmodium berghei-parasitized mice. International Journal of BCRR. 2016; 10(4):1-9.
Cizzy N, Engeu OP, Berna O, Norbert A, Esther M. Artemisia annua L. and Venonia amygdalina Del: A potential Herbal Artemisinin Combination Treatment against malaria. British Journal of Pharmaceutic Resources. 2016;14(3): 1-7.
Igbokwe VU, Eze DE, Adams DM, Kabiu KM, Ezekiel Ejeka PO, Okpara PO. Antimalarial effects of five traditional nigerian medicinal plant extracts on Plasmodium berghei-Infected Rats. FUDMA Journal of Sciences. 2021;5(2).
DOI: https://doi.org/10.33003/fjs - 2021 - 0502 - 461.
Soniran OT, Idowu OA, Ajayi OL, Olubi IC. Comparative study on the effects of Chloroquine and Artesunate on histopathological damage caused by Plasmodium berghei in four vital organs of infected albino mice. Journal of Malaria Research and Treatment, 2012;10: 11-55.
Idowu OA, Soniran, OT, Ajayi, OI, Olubi, IC. Effect of Morinda morindiode son organs of mice infected with Plasmodium berghei. Nigerian Journal of parasitology. 2014 35(1-2):31-40.
Elsheikha HM, Sheashaa HA. Epidemiology, pathophysiology, management and outcome of renal dysfunction associated with Plasmodium infection. Journal of Parasitology Research. 2007;101(5):1183-1190.
Bussayo AO, Laura Z, Olubusola DO, Oluwafunmike AS, Luciana D, Ademola CE. Ameliorative effects of ethanolic leaf extracts of Azadirachta indica on renal histologic alterations in streptozotocin-induced diabetic rats. The American Journal of Chinese Medicine. 2011;39(5):903-916.
Elias RM, Correa-Costa M, Barreto CR. Oxidative stress and modification of renal vascular permeability are associated with acute kidney injury during P. berghei ANKA infection. PLoS ONE. 2012;7(8): 440-444.
Koul A, Mohan V, Bharati S. Azadirachta indica mitigates DMBA-induced hepatotoxicity: a biochemical and radiometric study. Indian Journal of Biochemistry and biophysics. 2014;51(1): 37-45.
Anigboro AA, Onakurhefe P, Tonukari NJ, Avwioroko OJ, Egbeme E. Quantitative determination of some phytochemicals (phenol, flavonoid, saponin and alkaloid) in twenty-two Nigeria medicinal plants. Nigerian Journal of Science and Environment. 2018;13(1): 86-93.
Sato S. Plasmodium – a brief introduction to the parasites causing human malaria and their basic biology. Journal of Physiology/Anthropology. 2021;40:1-5.
Woldearegai TG, Lalremruata A, Nguyen IT, Gmeiner M, Veletzky L, Tazenda-kuitsouc GB. Characterisation of Plasmodium infections among inhabitants of rural areas in Gabon. Journal of Scientific Reports. 2019;9(1):41597-41599.
Kotepui M, Kotepui KU, Millanez GD, Masangkau FR. Severity and mortality of severe Plasmodium ovaleinfection: a systematic review and meta-analysis. PLoS ONE. 2020;15:e0235014.
Larson B, Barthelemy N, Stephan N, Nancy-Diamela M, Francois R, Virginie R, Frank P. Rodent Malaria in Garbon: Diversity and Host range. International Journal of Parasitology. 2019;10:117-124.
World Health Organization. World malaria report, Geneva; 2018.
Adeyemo-Salami OA, Farambi EO, Ademowo OG. An investigation into the antimalarial effect of methanolic extract of Paullinaipinnata leaves in P. berghei infected mice and course of infection. African Journal of Medicine and Medical Sciences. 2014;43(1):93-94.
World Health Organization/RBM. Documentation of the socio-economic impact of malaria epidermics in Africa. Final report. Available:www.rollbackmalaria.org.seim2019/(WHO/HTM/MAL/2019.1102).
Iwuchukwu IC, Vincent CN. Studies on prevalence of malaria and its adverse foetal outcomes in FMC, Owerri, Imo State, Nigeria.Archives of Community Medicine and Public Health. 2021; 7(2): 151-163.
Osonwa EU, Mbonu DO, Eluu CS, Oli AN. Antiplasmodial and biochemical effects of combination of ethanolic leave-extract of Azadirachta indicaand Ocimum gratissimum on Plasmodium berghei-infected mice. African Journal of Pharmaceutical Sciences and Pharmacy. 2017;5:1-6.
Afolabi OJ, Simon-oke IA, Oladokun OI. Antiplasmodial Activity of Ethanolic Extract of Neem Leaf (Azadirachta indica) in Albino Mice Infected with Plasmodium berghei. International Archives of Clinical Pharmacology. 2021;7:2572-2578.
Nwankwo EN, Ezekwesili OO, Chude CM. In vitro antiplasmodial and in vivo toxicity potentials of Mentha piperita and Ocimum gratissimum essential oils and their synergistic effect with conventional antimalarial drugs against Plasmodium falciparum. International Journal of Mosquito Research. 2022;9(1):114-122.
Omagha R, Idowu ET, Adeneye AK. Survey of ethnobotanical cocktails commonly used in the treatment of malaria in southwestern Nigeria. Future Journal of Pharmaceutical Sciences. 2021;7:152-158.
Nigussie G, Wale M. Medicinal plants used in traditional treatment of malaria in Ethiopia: a review of ethnomedicine, anti-malarial and toxicity studies. Malaria Journal. 2022;21:262-266.
Million E, Mulugeta T, Umeta B. Traditional medicine practice and its role in the management of malaria in Jimma Town, Oromia, Ethiopia. Journal of Infection and Drug Resistance. 2022;15:2187-2198.
Williamson, EM. Synergy and other interactions in Phytomedicines. Phytomedicine. 2001;8(5): 401-409.
Erhirhie, EO, Ikegbune, C, Okonkwo, OB. Antimalarial herbal drugs: a review of their interactions with conventional antimalarial drugs. International Journal of Phytomedicine and Physiotherapy. 2021;7:4-10.
Achi NK, Onyeabo C, Nnate DA, Ekeleme-Egedigwe CA, Kalu IK, Chibundu IC, Wokoma GC. Therapeutic effects of Azadirachta indica A. juss. leaves in malaria-induced male wister rats. Journal of Pharmacy and Pharmacognosy Research. 2018;6(3):191-204.
Mbugi EV, Sife AS, Mboni R, Grace EPM, Bestina D, Edda TL. Effectiveness of Azadirachta indica (neem tree) on prevention and treatment of clinical human malaria: A systematic review. Journal of East Africa Science. 2021;3:1-4.
Yarmohammadi F, Mehri S, Najan N, Salar AS, Hosseinzadeh, H. The protective effect of Azadirachta indica (Neem) against Metabolic Syndrome: A Review. Iran Journal of Basic Medical Sciences. 2021;24 (3):280-292.
Annette H, Barbara P, Sofia T, Judith N, Fabrizio, B. Effects of Azadirachta indica seed kernel extracts on early erythrocytic schizogony of Plasmodium berghei and proinflammatory response in inbred mice. Malaria Journal. 2019;18:35-43.
Alex DWA, Okorie PU. Water quality studies of Nworie Rivers in Owerri, Nigeria. Journal of Mississipi Academy of Sciences. 2008;20(1):55-60.
Ejebe DE, Emudianohwo JO, Ozoko T, Simialayi IM, Esume CO, Maduodi UV. An Investigation into the antiplasmodial effect of the ethanol extract of the leaves of Helliantusannus in Swiss Albino mice. Global Journal of Pharmacology. 2011;5 (2):92-96.
Peter IT, Anatoli VK. The current global malarial situation.Malaria parasite biology, pathogenesis, and protection. American Society for Microbiology, Washington DC. 1998;11-22.
Dawit D, Eyassu M, Asfaw D, Dawit A, Kelbessa U, Wallelign M, Daniel M, Ashenafi, A, yared, M. In vivo antimalaria activity of hydrocoholic extracts from Asparagus africanus Lam. In mice infected with Plasmodium berghei. Ethiopian Journal of Health Development. 2006; 20(2):112-118.
Ejebe DE, Siminialayi IM, Emudainowho JOT, omakporaye SL, Ojeih AE, Akonoghrere R, Odekuma IE, Ahatty GC. An improved technique for the oral administration of solutions of test substances to experimental rats using mediflon.medicut intravenous cannula. American Journal of Biotechnology. 2009; 8(6):60-96.
Lorke D. A New Approach to Practical Acute Toxicity Testing. Achives of Toxicology. 1983;54:275-287.
Matsumura, F. Toxicology of insecticides. Plenum press, York and London. 1975; 24.
Corbett JR, Wright, K, Baillie, AC. The Biochemical mode of Action of pesticides, 2nd, Academic press, London and New York; 1984.
David AF, Lip JR, Simon LC, Rato B, Solomon N. Antimalarial drug discovery: efficacy models for compound screening. Nature review. 2004;16:522-528.
Awe SO, Makinde, JM. Effect of petroleum ether fractions of Morindalucida on Plasmodium bergheiberghei in mice. Journal of Pharmaceutical Biology. 1998;36(4):301-304.
Tona L, Mesia, K, Ngimbi NP, Chrimwami,B, Okond A, Cimanga K, Bruyne, TD, Apers, S, Hermans, N, Totte, J, Pieters, L. In-vivo antimalarial activity of Cassia occidentalis, Morinda morindoide sand Phyllanthusniruri. Annals of Tropical Medicine and Parasitology. 2001;95(1) :47-57.
Iwalokun BA. Enhanced antimalarial effects of chloroquine by aqueous Vernonia amygdalina leaf extract in mice infected with chloroquine resistant and sensitive Plasmodium berghei strains. Journal of African Health Sciences. 2008; 8(1):25-35.
Adebajo, AC, Odediran, SA, Aliyu, AF, Nwafo, PA, Nwoko, TN, Umana, SU. In Vivo Antiplasmodium Potentials of the Combinations of four Nigerian Antimalarial Plants. Journal of Molecules. 2014;19(9): 13136-13146.
Somsak V, Chachiyo S, Kittitorn J, Audomkasok S, Sriwiphat S. Antimalarial and Antihypoglycemic Properties of Siamese Neem Tree (Azadirachta indica) in Plasmodium berghei-infected Mice. Malaria Contraction Elimination. 2015;4: 134-137.
Ekpo DE, Ekanemesang MU. Antiplasmodial/antimarlarial effect of ethanol extracts of leaves of Vernonia amygdalina and Gongronema latifolium on the activity of catalase in Plasmodium berghei-parasitized mice. International Journal of BCRR. 2016; 10(4):1-9.
Cizzy N, Engeu OP, Berna O, Norbert A, Esther M. Artemisia annua L. and Venonia amygdalina Del: A potential Herbal Artemisinin Combination Treatment against malaria. British Journal of Pharmaceutic Resources. 2016;14(3): 1-7.
Igbokwe VU, Eze DE, Adams DM, Kabiu KM, Ezekiel Ejeka PO, Okpara PO. Antimalarial effects of five traditional nigerian medicinal plant extracts on Plasmodium berghei-Infected Rats. FUDMA Journal of Sciences. 2021;5(2).
DOI: https://doi.org/10.33003/fjs - 2021 - 0502 - 461.
Soniran OT, Idowu OA, Ajayi OL, Olubi IC. Comparative study on the effects of Chloroquine and Artesunate on histopathological damage caused by Plasmodium berghei in four vital organs of infected albino mice. Journal of Malaria Research and Treatment, 2012;10: 11-55.
Idowu OA, Soniran, OT, Ajayi, OI, Olubi, IC. Effect of Morinda morindiode son organs of mice infected with Plasmodium berghei. Nigerian Journal of parasitology. 2014 35(1-2):31-40.
Elsheikha HM, Sheashaa HA. Epidemiology, pathophysiology, management and outcome of renal dysfunction associated with Plasmodium infection. Journal of Parasitology Research. 2007;101(5):1183-1190.
Bussayo AO, Laura Z, Olubusola DO, Oluwafunmike AS, Luciana D, Ademola CE. Ameliorative effects of ethanolic leaf extracts of Azadirachta indica on renal histologic alterations in streptozotocin-induced diabetic rats. The American Journal of Chinese Medicine. 2011;39(5):903-916.
Elias RM, Correa-Costa M, Barreto CR. Oxidative stress and modification of renal vascular permeability are associated with acute kidney injury during P. berghei ANKA infection. PLoS ONE. 2012;7(8): 440-444.
Koul A, Mohan V, Bharati S. Azadirachta indica mitigates DMBA-induced hepatotoxicity: a biochemical and radiometric study. Indian Journal of Biochemistry and biophysics. 2014;51(1): 37-45.
Anigboro AA, Onakurhefe P, Tonukari NJ, Avwioroko OJ, Egbeme E. Quantitative determination of some phytochemicals (phenol, flavonoid, saponin and alkaloid) in twenty-two Nigeria medicinal plants. Nigerian Journal of Science and Environment. 2018;13(1): 86-93.
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