Anthelminthic Activity of Moringa oleifera Leaf Extracts Evaluated in Vitro on Four Developmental Stages of Haemonchus contortus from Goats

Gertrude Mbogning Tayo, Josué Wabo Poné, Marie Claire Komtangi, Jeannette Yondo, Alidou Marc Ngangout, Mpoame Mbida

Department of Animal Biology, Faculty of Science, University of Dschang, Dschang, Cameroon.

DOI: 10.4236/ajps.2014.511185                

Abstract

Haemonchus contortus is a blood-sucking abomasal helminth of small ruminants responsible for major economic losses to small farmers worldwide. Widespread resistance to synthetic anthelminthics has stimulated a need for alternative strategies of parasite control, among which is the use of medicinal plants with natural anthelminthic properties. This study assessed in vitro the efficacy of infused and macerated aqueous extract as well ethanolic extract of Moringa oleifera against fresh eggs, embryonated eggs, L₁ and L₂ larvae of H. contortus. For this purpose, five different concentrations (0.625, 1.25, 2.5, 3.75 and 5 mg/ml) were prepared from dry extracts via serial dilutions with distilled water. Fresh eggs obtained from artificially infected goat feces were exposed to these different concentrations for 48 hours, while embryonated eggs and larvae were exposed for 6 and 24 hours respectively. Distilled water and 1.5% DMSO were used as negative control. The results were expressed in terms of mean inhibition percentage of egg embryonation, mean inhibition percentage of egg hatch and mean percentage of larval mortality. An overview of results revealed that ethanolic leaf extract of M. oleifera was most efficient on eggs by inhibiting 60.3% ± 8.2% and 92.8% ± 6.2% eggs embryonation at 3.75 and 5 mg/ml respectively with a significant difference (P < 0.05), which contributed to obtaining the lowest LC₅₀ value of 0.985 mg/ml. This extract also inhibited 99% ± 2% egg hatching of H. contortus at 5 mg/ml with an LC₅₀ value of 1.7 mg/ml. Concerning activity on larvae, the ethanolic extract was also most potent against them by inducing 98.8% ± 2.5% and 100% ± 0% mortality of L₁ and L₂ larvae at 5 mg/ml respectively. Infused aqueous extract was more efficient on eggs than on larvae with an IC₅₀ value less than 2 mg/ml and an LC₅₀ value more than 3.5 mg/ml. Macerated aqueous extract showed good activity against the four developmental stages with LC₅₀ values ranging from 2.08 mg/ml for L₂ larvae to 2.92 mg/ml for L₁ larvae and 2.37 to 2.52 mg/ml for embryonated and fresh eggs respectively. The current study showed that all three extracts of M. oleifera tested possessed potential ovicidal and larvicidal activities against H. contortus. However, further in vivo studies are necessary to validate the anthelminthic property of this plant.

Keywords

Moringa oleifera, Ovicidal, Larvicidal, Haemonchus contortus, Goats, Cameroon

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Sujon, M.A., Mostoja, M., Jahan, M.S., Das, A.R. and Rob, S. (2008) Studies on Medicinal Plants against Gastrointestinal Nematodes of Goats. Bangladesh Journal of Veterinary Medicine, 6, 179-183.
[2]	Sabater, M. (2012) Détermination d’une dose efficace et d’une dose toxique de tanins condensés dans le contrôle des strongyloses digestives chez les Caprins. Thèse d’exercice, Médecine vétérinaire, Ecole National Vétérinaire de Toulouse—ENVT, Toulouse, 135p.
[3]	Labonne, M., Magrong, P. and Oustalet, Y. (2002) Le secteur de l’élévage au Cameroun et dans les provinces du grand Nord: Situation actuelle, contraintes, enjeux et défis. Acte du colloque, Garoua, 27-31 Mai 2002.
[4]	Botura, M.B., Silva, G.D., Lima, H.G., Oliveira, J.V.A., Souza, T.S., Santos, J.D.G., Branco, A., Moreira, E.L.T., Almeida, M.A.O. and Batatinha, M.J.M. (2011) In Vivo Anthelminthic Activity of an Aqueous Extract from Sisal Waste (Agave sisalana Perr.) against Gastro-Intestinal Nematodes in Goats. Veterinary Parasitology, 177, 104-110. http://dx.doi.org/10.1016/j.vetpar.2010.11.039
[5]	Urquhart, G.M., Armour, J., Duncan, J.L., Dunn, A.M. and Jennings, F.W. (1996) Veterinary Parasitology. 2nd Edition, Blackwell Science Ltd., Oxford, 224-234.
[6]	Kamaraj, C. and Abdul Rahuman, A. (2011) Efficacy of Anthelmintic Properties of Medicinal Plant Extracts against Haemonchus contortus. Research on Veterinary Science, 91, 400-404. http://dx.doi.org/10.1016/j.rvsc.2010.09.018
[7]	Hafiz, A.B., Zafar, I., Muhammad, N.K., Zia-ud-Din, S. and Abdul, J. (2009) Anthelmintic Activity of Ziziphus nummularia (Back) and Acacia nilotica (fruit) against Trichostrongylid Nematodes of Sheep. Journal of Ethnopharmacology, 123, 325-329. http://dx.doi.org/10.1016/j.jep.2009.02.043
[8]	Devendra, B.N., Srinivas, N., Talluri, P.S.S.L.P. and Latha, P.S. (2011) Antimicrobial Activity of Moringa oleifera Lam., Leaf Extract, against Selected Bacterial and Fungal Strains. International Journal of Pharma and Bio Sciences, 2, 13-18.
[9]	Wasswa, P and Deogracious, O. (2005) The in Vitro Ascaridal Activity of Selected Indigenous Medicinal Plants Used in Ethno-Veterinary Practices in Uganda. African Journal of Traditional, Complementary and Alternative Medicines, 3, 94-103.
[10]	Chandrashekar, K.S., Thakur, A. and Prasanna, K.S. (2010) Anti-Inflammatory Activity of Moringa oleifera Stem Back Extracts against Carrageenen Induced Rat Paw Edema. Journal of Chemical and Pharmaceutical Research, 2, 179-181.
[11]	Moya, B., Oyedemi, S., Masika, P.J. and Muchenje, V. (2012) Polyphenolic Content and Antioxydant Properties of Moringa oleifera Leaf Extracts and Enzymatic Activity of Liver from Goats Supplemented with Moringa oleifera Leaves/Sun Flower Seed Cake. Meat Science, 91, 441-447. http://dx.doi.org/10.1016/j.meatsci.2012.02.029
[12]	Singh, N., Verna, V.K., Saxena, P. and Singh, R. (2012) Anti-Ulcer and Antioxidant Activity of Moringa oleifera (Lam) Leaves against Aspirin and Ethanol Induced Gastric Ulcer in Rats. International Research Journal of Pharmaceuticals, 2, 46-57.
[13]	Mbikay, M. (2012) Therapeutic Potential of Moringa oleifera Leaves in Chronic Hyper-Glycemicemia and Dyslipidemia: A Review. Frontiers in Pharmacology, 3, 1-37.
[14]	Wabo Poné, J., Bilong Bilong, C.F., Mpoame, M., Catherine, F.N. and Coles, C.C. (2006) In Vitro Activity of Ethanol, Cold and Hot Water Extracts of the Stem Back of canthium mannii (Rubiaceae) on Ancylostoma caninum Eggs. East and Central African Journal of Pharmaceutical Sciences, 9, 14-18.
[15]	Wabo Poné, J., Bilong Bilong, C.F. and Mpoame, M. (2010) In Vitro Nematicidal Activity of Extracts of Canthium mannii (Rubiaceae) on Different Life Cycle of Heligmosomoides polygyrus (Nematoda, Heligmosomatidae). Journal of Helminthology, 84, 156-165. http://dx.doi.org/10.1017/S0022149X09990435
[16]	Mbogning Tayo, G., Payne, V.K., Wabo Poné, J., Komtangi, M.C., Yondo, J., Ngangout, A.M., Mpoame, M. and Bilong Bilong, C.F. (2012) In Vitro Ovicidal and Larvicidal Activities of Aqueous and Ethanolic Extracts of the Leaves of Bidens pilosa (Asteraceae) on Heligmosomoides bakeri (Nematoda: Heligmosomatidae). Medicinal Plants, 4, 121-125.
[17]	Wabo Poné, J., Olivia Fossi, T., Yondo, J., Komtangi, C.M., Mpoame, M. and Bilong Bilong, C.F. (2011) The in Vitro Effects of Aqueous and Ethanolic Extracts of the Leaves of Ageratum conyzoides (Asteraceae) on Three Life Cycle Stages of the Parasitic Nematode Heligmosomoides bakeri, (Nematoda: Heligmosomatidae). Veterinary Medicine International, 2011, Article ID: 140293.
[18]	Wabo Poné, J., Ngankam, N.J.D., Bilong Bilong, C.F. and Mpoame, M. (2011) A Comparative Study of the Ovicidal and Larvicidal Activities of Aqueous and Ethanolic Extracts of Pawpaw Seed Carica papaya (Caricaceae) on Heligmosomoides bakeri. Asian Pacific Journal of Tropical Medicine, 4, 447-450.
[19]	Katiki, L.M., Chagas, A.C.S., Bizzo, H.R., Ferreira, J.F.S. and Amarante, A.F.T. (2011) Anthelmintic Activity of Cymbopogon martini, Cymbopogon schoenanthus and Mentha piperita Essential Oils Evaluated in Four Different in Vitro Tests. Veterinary Parasitology, 183, 103-106. http://dx.doi.org/10.1016/j.vetpar.2011.07.001
[20]	Asase, A., Oting-Yeboah, A.A., Odamtten, G.T. and Simmonds, M.S.J. (2005) Ethnobotanical Study of Some Ghanaian Anti-Malarial Plants. Journal of Ethnopharmacology, 99, 273-279. http://dx.doi.org/10.1016/j.jep.2005.02.020
[21]	Tatik, S. and Dwatmadji (2011) Anthelmintic Activity of Melastoma malabatricum Extract on Haemonchus contortus Activity in Vitro. Asian Journal of Pharmaceutical and Clinical Research, 4, 68-70.
[22]	Soulsby, E.J.L. (1982) Helminths, Arthropods and Protozoa of Domesticated Animals. 7th Edition, Bailler, Tindall, London.
[23]	Wabo Poné, J., Payne, V.K., Mbogning, T.G., Komtangi, M.C., Yondo, J., Ngangout, A.M., Mbida, M. and Bilong Bilong, C.F. (2013) In Vitro Anthelminthic Efficacy of Dichrocephala integrifolia (Asteraceae) Extracts on the Gastro-Intestinal Nematode Parasite of Mice: Heligmosomoïdes bakeri (Nematoda, Heligmosomatidae). Asian Pacific Journal of Tropical Biomedicine, 3, 100-104. http://dx.doi.org/10.1016/S2221-1691(13)60032-5
[24]	Wabo Poné, J., Kenné Tameli, F., Mpoame, M., Pamo, T.E. and Bilong Bilong, C.F. (2011d) In Vitro Activities of Acetonic Extracts from Leaves of Three Forage Legumes (Calliandra calotyrsus, Gliricidia sepium and Leucaena diversifolia) on Haemonchus contortus. Asian Pacific Journal of Tropical Medicine, 4, 125-128.
[25]	Rastogi, T., Bhutda, V., Moon, K., Aswar, P.B. and Khadabadi, S.S. (2009) Comparative Studies on Anthelmintic Activity of Moringa oleifera and Vitex negundo. Asian Journal of Research in Chemistry, 2, 181-182.