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In vitro nematicidal activity of extracts of Canthium mannii (Rubiaceae), on different life-cycle stages of Heligmosomoides polygyrus (Nematoda, Heligmosomatidae)

  • J. Wabo Poné (a1), C.F. Bilong Bilong (a2) and M. Mpoame (a1)

The increasing prevalence of anthelmintic-resistant strains of helminths, drug residues in animal products and high cost of conventional anthelmintics has created an interest in studying medicinal plants as an alternative source of anthelmintic. The potential nematicidal activities of four extracts from the bark of Canthium mannii (Rubiaceae) stem were investigated in vitro. Extracts were diluted in distilled water (DW) to obtain five different concentrations (1.5, 2.0, 2.5, 3.0 and 3.5 mg/ml) and put in contact with eggs and larvae of Heligmosomoides polygyrus. The different stages of the life cycle were also put in contact with the same concentration of mebendazole (MBZ, positive control). One millilitre of each extract at different concentrations and control were added to 1 ml solution containing 30–40 eggs or 10–15 larvae (L1, L2 and L3) and distributed in different Petri dishes. The eggs and larvae were incubated at 24°C and exposure times were: 48 h for un-embryonated eggs, 6 h for embryonated eggs; 2, 4, 6 and 24 h for L1 and L2 larvae, 24–48 h for infective larvae (L3), and 5 days for the larval development test (from L1 to L3). DW and 1% dimethyl sulphoxide (DMSO) were used as placebo and DMSO control, respectively. Significant effects were obtained with three of the four extracts, and differences were observed depending on the parasite stage. Cold water extract (CWE), hot water extract (HWE) and ethanol extract (ETE) inhibited embryonic development (40, 45 and 10%) and hatching of embryonated eggs (40, 85 and 80%), respectively, at 3.5 mg/ml. Only ETE killed L1 (97.18%) and L2 (92.68%) larvae of H. polygyrus after 24 h at 3.5 mg/ml and drastically reduced the production rate (6% at 3.0 and 3.5 mg/ml) of infective larvae (L3) after 5 days of incubation compared to other extracts (P < 0.05). However, the infective larvae of H. polygyrus were resistant to the effect of each of the tested products (extracts and mebendazole). These in vitro results suggested that extracts of C. mannii, used by traditional healers in Dschang, Western Region of Cameroon (Central Africa) to cure intestinal helminthiasis and abdominal pains of their patients, possess nematicidal properties. The active principles responsible for the activity could be secondary metabolites such as alkaloids and saponins present in the extracts. It is suggested that further experiments incorporating in vivo purification of extracts and toxicological investigations should be carried out.

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Ademola, I.O. (2002) A survey on ivermectin resistance in strongyles of sheep in Oyo state Nigeria using larval development. Israeli Journal of Veterinary Medicine 54, 247252.
Adjanohoun, J.E., Aboubakar, N., Dramame, K., Ebat, M.E., Ekpére, J.A., Enoworock, E.G., Focho, D., Cibile, Z.O., Kamanyi, A., Kamsu kom, J., Keita, A., Mbenkum, T., Mbi, L.N., Mbiele, A.L., Mbomé, L.C., Mubiri, W.K., Nancy, W.L., Kongmeneck, B., Satabie, B., Sofovora, A., Tamze, V. & Wirmum, C.K. (1995) Traditional medicine and pharmacopoeia. Contribution to ethnobotanica and floristic studies in Cameroon. 569 pp. Yaoundé, Cameroun, Centre National de Production de Manuels Scolaires (CNPMS).
Alawa, C.B.F., Adamu, A.M., Gefu, J.O., Ajanusi, O.J., Abdu, P.A., Chiezey, N.P., Alawa, J.N. & Bowman, D.D. (2003) In vitro screening of two medicinal plants (Vernonia amygdalina and Annona senegalensis) for anthelmintic activity. Veterinary Parasitology 1, 7381.
Assis, L.M., Bevilaqua, C.M.L.S., Moraisa, M., Vieirab, L.S., Costar, C.T.C. & Souzaa, J.A.L. (2003) Ovicidal and larvicidal activity in vitro of Spigelia anthelmia extracts on Haemonchus contortus. Veterinary Parasitology 117, 4344.
Balansard, G., Maillard, C. & Vidal-Ollivier, E. (1991) Méthodes d'extraction en vue d'obtenir des extraits ou produits purs à partir des plantes médicinales. 18 pp. Paris, France, Editions Techniques. Phytothérapie, Aromathérapie.
Boisvenue, R.J., Brand, M.C., Galloway, R.B. & Hendrix, J.C. (1983) In vitro activity of various anthelmintic compounds against Haemonchus contortus larvae. Veterinary Parasitology 13, 341347.
Busindou, M. (1997) Enquête sur la phytothérapie traditionnelle à Kindamba et Odzala (Congo) et analyse des convergences d'usage des plantes médicinales en Afrique centrale. 482 pp. Doctoral thesis, Université Libre de Bruxelles.
Cheesbrough, M. (1987) Medical laboratory manual for tropical countries. 2nd edn. 605 pp. Cambridge, Cambridge University Press.
Cirak, U.Y., Crulegen, E. & Baner, C. (2004) Benzimidazole resistance in Cyathosmin populations on horse farm in western Anatobia, Turkey. Parasitology Research 93, 392395.
Ciulei, I. (1982) Methodology for analysis of vegetable drugs, practical and aromatic plants. 67 pp. Bucharest, Romania.
Dobson, R.J., Donal, A.D., Waller, P.J. & Snowdon, K.L. (1986) An egg-hatch assay for resistance to levamisole in trichostrongyloid nematode parasites. Veterinary Parasitology 19, 7784.
Dupouy-Camet, J. (2000) Le traitement de la trichinellose. Pyrexie 4, 117121.
Elard, L. (2000) La résistance aux benzimidazoles chez Tladorsagia circumcincta, nématode parasite de petits ruminants. Etude du déterminisme génétique et recherches des conséquences sur la fitness des parasites. 200 pp. Doctoral thesis, Université de Tours, Paris.
Euzeby, J. (1981) Diagnostic expérimental des helminthoses animales (animaux domestiques, animaux de laboratoire, Primate). Travaux pratiques d'helminthologie vétérinaire. Livre 1 généralités. Diagnostic ante mortem. 349 pp. Paris, Edition ‘informations techniques des services vétérinaires’.
Githiori, J.B. (2004) Evaluation of anthelmintic properties of ethnoveterinary plant preparations used as livestock dewormers by pastoralist and small holder farmers in Kenya. 76 pp. Doctoral dissertation, Department of Biomedical Sciences and Veterinary Public Health, Acta Universitatis Agricultural sulciae.
Githiori, J.B., Hoglund, J., Waller, P.J. & Baker, R.L. (2003a) The anthelmintic efficacy of the plant, Albizia anthelmintica, against the nematode parasites Heamonchus contortus of sheep and Heligmosomoides polygyrus of mice. Veterinary Parasitology 116, 2334.
Githiori, J.B., Hoglund, J., Waller, P.J. & Baker, R.L. (2003b) Evaluation of anthelmintic properties of extracts from some plants used as livestock dewormers by pastoralist and small holder farmers in Kenya against Heligmosomoides polygyrus infections in mice. Veterinary Parasitolology 118, 215226.
Gunasegaran, R., Subramani, K., Azantha Parimala, P., Ramachandra Nair, A.G., Rodriguez, C.T.C. & Souzaa, J.A.L. (2003) Ovicidal and larvicidal activity in vitro of Spigelia anthelmia extracts on Haemonchus contortus. Veterinary Parasitology 117, 4344.
Hubert, J. & Kerboeuf, D. (1992) A microlarval development assay for the detection of anthelmintic resistance in sheep nemode. Veterinary Record 130, 442446.
Ibarra, O.F. & Jenkins, D.C. (1984) The relevance of in vitro anthelmintic screening tests employing the free-living stages of trichostrongylid nematodes. Journal of Helmintholology 58, 107112.
Kanchanapoom, T., Kasai, R. & Yamasaki, K. (2002) Iridoid and phenolic diglycosides from Canthium berberidifolium. Phytochemistry 61, 461464.
Lans, C. & Brown, G. (1998) Ethnoveterinary medicines used for ruminants in Trinidad and Tobago. Preventive Veterinary Medicine 35, 149163.
Magdeleine, M.C., Hoste, H., Mathieu, M., Varo, H. & Archimède, H. (2009) In vitro effects of Cucurbita moschata extracts on Haemonchus contortus. Veterinary Parasitology 161, 99105.
Matthee, S., Dreyer, F.H., Hoffmann, W.A. & Van Niekerk, F.E. (2002) An introductory survey of helminth control practice in South Africa and anthelmintic resistance on thoroughbred farms in the western Cape Province. Journal of the South Africa Veterinary Association 73, 195200.
McGaw, L., Jager, A.K. & Van Staden, J. (2000) Antibacterial, anthelmintic and antiamoebic activity in South African medicinal plants. Journal of Ethnopharmacology 72, 247263.
Michael, B., Meinke, T.P. & Shoop, W. (2001) Comparison of ivermectin, doramectin, selamectin and eleven intermediates in a nematode larval development assay. Journal of Parasitology 87, 692696.
Mpoame, M. & Essomba, I. (2000) Essai de traitement contre des parasitoses gastro-intestinales du poulet avec les décoctions aqueuses de graines de papaye (Carica papaya). Revue d'Elevage et de Médecine Vétérinaire des Pays Tropicaux 53, 2325.
Ndao, M., Belot, J., Zinsstag, J. & Pfister, K. (1995) Epidémiologie des helminthiases gastro-intestinales des petits ruminants dans la zone sylvo-pastorale au Sénégal. Veterinary Record 36, 132139.
Noritaka, H., Tavo, O. & Viazulito, H. (2000) Challenge trial on the anthelmintic effect of drugs and natural agents against the monogenean Heterobothrium okamotoi in the tiger puffer Tokipugu rubripes. Aquaculture 18, 113.
Oliveira, L.M.B., Bevilaqua, C.M.L., Costa, C.T.C., Macedo, I.T.F., Barros, R.S., Rodrigues, A.C.M., Cumurça-Vasconcelos, A.L.F., Morais, S.M., Lima, Y.C., Vieira, L.S. & Navarro, A.M.C. (2009) Anthelmintic activity of Cocos nucifera L. against sheep gastrointestinal nematodes. Veterinary Parasitology 159, 5559.
Pessoa, L.M., Movais, S.M., Bevilaqua, C.M.L. & Luciano, J.H.S. (2002) Anthelmintic activity of essential oil of Ocimum gratissimum L. and euguenol against Heamonchus. Veterinary Parasitology 24, 15.
Quilan, M.B., Quinlan, R.J. & Nolan, J.M. (2002) Ethnophysiology and herbal treatments of intestinal worms in Dominica, West Indies. Journal of Ethnopharmacolology 103, 183194.
Sangster, N.C. (1999) Anthelmintic resistance: past, present and future. International Journal of Parasitology 29, 115124.
Satrija, F., Retnani, E.B., Ridwan, Y. & Tiuria, R. (2001) Potential use of herbal anthelmintics as alternative antiparasitic drugs for small holders in developing countries. Proceedings of the 10th Conference of the Association of Institutions for Tropical Veterinary Medicine. 20–23 August. Copenhagen, Denmark.
Sinha, H.K., Sivastava, P.S., Singh, S.P., Sinha, K.V. & Sinha, S.R. (1987) Efficacy of various anthelmintics on the mortality of the infective larva of Toxocara vitulorum and treatment of calf ascariasis. Indian Journal of Animal Science 57, 185188.
Smyth, J.D. (1996) Animal parasitology. 549 pp. London, Cambridge University Press.
Tariq, K.A., Chishti, M.Z., Ahmad, F. & Shawl, A.S. (2009) In vitro effects of Cucurbita moschata seed extracts on Haemonchus contortus. Veterinary Parasitology 161, 99105.
Taylor, M.A. (1990) A larval development test for the detection of anthelmintic resistance in nematodes of sheep. Research Veterinary Science 49, 198202.
Taylor, M.A., Hunt, K.R. & Goodyear, K.L. (2002) Anthelmintic resistance detection methods. Veterinary Parasitology 103, 183194.
Wabo Poné, J. (2001) Etude comparée in vitro de l'activité anthelminthique de l'extrait éthanolique de la poudre d'écorce de Canthium mannii (Rubiaceae) et du telkan. 45 pp. Cameroun, Mémoire de DEA, Département de Biologie et Physiologie Animales, Université de Yaoundé 1.
Wabo Poné, J. (2007) Effets des extraits de Canthium mannii Hiern, 1877 (Rubiacées) sur deux nématodes parasites strongylides: Ancylostoma caninum Ercolani, 1859 et Heligmosomoides polygyrus Dujardin, 1845. 155 pp. Unpublished PhD thesis, Département de Biologie et Physiologie Animales, Faculté des Sciences, Université de Yaoundé 1, Cameroun.
Wabo Poné, J., Mpoame, M., Bilong Bilong, C.F. & Kerboeuf, D. (2005) Etude comparée in vitro de l'activité anthelminthique de l'extrait éthanolique de la poudre d'écorce de Canthium mannii (Rubiaceae) et du Mébendazole. Revue de Médecine Vétérinaire 15, 633636.
Wabo Poné, J., Bilong Bilong, C.F., Mpoame, M., Fusi-Ngwa, C. & Coles, C.G. (2006) In vitro activity of ethanol, cold and hot water extracts of the stem bark of Canthium mannii (Rubiaceae) on Ancylostoma caninum eggs. East and Central African Journal of Pharmaceutical Sciences 9, 1418.
Wolstenholme, A.J., Fairweather, I., Prichard, R., Von Samson-Himmelstjerna, G. & Songster, N.C. (2004) Drug resistance in veterinary helminths. Trends in Parasitolology 20, 469476.
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Journal of Helminthology
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  • EISSN: 1475-2697
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