In Vitro and in Vivo Anthelmintic Activities of Aqueous Leaf Infusion of Azadirachta indica against Haemonchus contortus

A. A. Sakti, Kustantinah Kustantinah, R. W. Nurcahyo


Anthelmintic resistance of Haemonchus contortus worm has become a major problem in ruminant production. Traditional medicinal and potential tropical plants with high tannin contents have a great potency as an alternative source of bio-anthelmintic. The study was carried out to assess the in vitro egg hatches inhibiting (EHI) and adult worm motility (AWM) tests and in vivo anthelmintic activities of aqueous leaf infusion of Azadirachta indica against H. contortus. Leaf infusion at doses of 2%, 4%, and 6% were used for in vitro treatments, and albendazole at a dose of 2 mg/mL was used as a positive control and 0.9% of sodium chloride was used as a negative control. The EHI assay was conducted two times i.e., before treatment and 24 h after treatment. The AWM were monitored 15 and 30 min, and 1, 2, 3, 4, 5, 6, 7, and 8 h post treatment. Sixteen Javanese Thin Tail ewes at the ages of ±12 months that naturally positive of H. contortus with about 1.000 EPG were selected and allocated randomly to 4 groups (n= 4). They were subsequently received the ensuing treatments i.e., A. indica infusion at doses of 0% (A) as a control, 6% (B), 8% (C), and 5 mg/BW albendazole (D) at a single oral dose as a positive control. Experimental ewes in Groups B and C were given oral infusion weekly during 4 weeks of treatment. EPG’s fecal examinations were conducted weekly. The result showed that the leaf infusion of A. indica containing condensed tannin (CT) at a dose of 6% significantly inhibited egg hatching (P<0.05) with a better effect compared to doses of 0%, 2%, and 4%. In AWM test, three doses of infusion significantly difference each other, as well as both negative and positive controls (P<0.05). After treatment, both of the A. indica containing CT levels and albendazole reduced EPG count significantly (P<0.05). Meanwhile, significant difference was not detected in dose of 8% aqueous leaf infusion of A. indica and albendazole treatments on weeks 3, 4, 5, and 6 consecutively. In conclusion, the aqueous leaf infusion of A. indica is fully potential as a bio anthelmintic against H. contortus.


Haemonchus contortus; Azadirachta indica; anthelmintic activity; leaf extract; sheep

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Abdulrazak, S. A. & T. Fujihara. 1999. Animal Nutrition: A Laboratory Manual. Laboratory of Animal Science. Faculty of Life and Enviromental Science. Shimane University. Japan. 1: 24 – 28.

Adjorlolo, L. K., E. C. Timpong-Jones, S. Boadu, & T. Adogla-Bessa. 2016. Potential contribution of neem (Azadirachta indica) leaves to dry season feeding of ruminants in West Africa. Livest. Research R. Dev. 28: 1-9.

Athanasiadou, S., I. Kyriazakis, F. Jackson, & R. L. Coop. 2001. The effect of condensed tannins supplementation of foods with different protein content of parasitism, food intake, and performance of sheep infected with Trichostrongylus colubriformis. British J. Nut. 86: 697–706.

AOAC. 2005. Official Methods of Analysis of AOAC International. 18th ed. Assoc. Off. Anal. Chem., Arlington.

Beriajaya, J. M. & D. Haryuningtyas. 2005. Efficacy of pineaaple skin extract to control Haemonchus contortus on Sheep. Husbandry Technology and Veteriner National Seminar. 934-940.

Besier, R. B., L. P. Khan, N. D. Sargison, & J. A. Van Wyk. 2016. Diagnosis, treatment and management of Haemonchus contortus in small ruminants. Chapter Six. Adv. Parasitol. 93: 181-238.

Budisatria, I. G. S., H. M. J. Udo, C. H. A. M. Eilers, & A. J. van der Zijpp. 2007. Dynamic of small ruminant production a case study of Central Java, Indonesia. Agriculture. 36: 145–152.

Coles, G. C., C. Bauer, F. H. M. Borgsteede, S. Geerts, T. R. Klei, M. A. Taylor & P. J. Waller. 1992. World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) methods for the detection of anthelmintic resistance in nematodes of veterinary importance. Vet. Parasitol. 44: 35-44.

Coles, G. C., F. Jackson, W. E. Pomroy, R. K. Prichard, G. Von Samson-Himmelstjerna, A. Silvestre, M. A. Taylor, & J. Vercruysse. 2006. The detection of anthelmintic resistance in nematodes of veterinary importance. Vet. Parasitol. 136: 167-185.

Costa, C. T. C., C. M. L. Bevilaqua, A. L. F. Camurça-Vasconcelos, M. V. Maciel, S. M. Morais, C. M. C. Castro, R. R. Braga, L. M. B. Oliveira. 2008. In vitro ovicidal and larvacidal activity of Azadirachta indica extracts on Haemonchus contortus. S. Rum. Research. 74: 284-287.

Eguale, T., G. Tilahun, A. Debella, A. Feleke, & E. Makonnen. 2007. In vitro and in vivo anthelmintic activity of crude extracts of Coriandrum sativum against Haemonchus contortus. J. Ethnophar. 110: 428-433.

Ferreira, L. E., P. M. N. Castro, A. C. S. Chagas, S. C. França, & R. O. Beleboni. 2013. In vitro anthelmintic activity of aqueous leaf extract of Annona muricata L. (Annonaceae) against Haemonchus contortus from sheep. Exp. Parasitol. 134: 327-332.

Gasser, R.B., E.M. Schwarz, P.K. Korhonen, & N.D. Young. 2016. Understanding Haemonchus contortus better through genomics and transcriptomics. Adv. Parasitol. 93: 519-547.

Gilleard, J. S. 2006. Understanding anthelmintic resistance: the need for genomics and genetics. Int. J. Parasitol. 36: 1227-1239.

Hamad, K. K., Z. Iqbal, Z. D. Sindhu, R. Z. Abbas, A. Khan, G. Muhammad, & B. Epperson. 2013. Combination of Nicotiana tabacum and Azadirachta indica: a novel substitute to control levamisole and ivermectin-resistant Haemonchus contortus in ovine. Pak. Vet. J. 34: 24-29.

Hoste, H., F. Jackson, S. Athanasiadou, S. M. Tamsborg, & S.O. Hoskin. 2006. The effect of tannin-rich plants on parasitic nematodes in ruminants. Trends Parasitol. 22: 253-261.

Hoste, H., J. F. J. Torres-Acosta, J. Quijada, I. Chan-Perez, M. M. Dakheel, D. S. Kommuru, I. Muller-Harvey, & T. H. Terrill. 2016. Interactions Between Nutrition and Infections with Haemonchus contortus and Related Gastrointestinal Nematodes in Small Ruminants. Adv. Parasitol. 93: 239-351.

Hoste, H., J. F. J. Torres-Acosta, C. A. Sandoval-Castro, I. Mueller-Harvey, S. Sotiraki, H. Louvandini, S. M. Thamsborg, & T. H. Terrill. 2015. Tannin containing legumes as a model for nutraceuticals against digestive parasites in livestock. Vet. Parasitol. 212: 5-17.

Jamra, N., G. Das, P. Singh, & M. Haque. 2014. Anthelmintic efficacy of crude neem (Azadirachta indica) leaf powder against bovine strongylosis. J. Parasitic Diseases. 39: 786-788.

Jayanegara, A., M. Ridla, D. A. Astuti, K. G. Wiryawan, E. B. Laconi, & Nahrowi. 2017. Determination of energy and protein requirements of sheep in Indonesia using meta-analytical approach. Med. Pet. 40: 124-133.

Kerboeuf, D., M. Riou, & F. Guegnard. 2008. Flavonoid and Related Compounds in Parasitic Disease Control. Mini. Rev. Med. Chem. 8: 116-128.

Maherisis, N., M. C. Semsari, B. Eshratkhah, M. Sadaghian, A. Gorbani & S. Hassanpour. 2011. Evaluation of the effects of Quebracho condensed tannin on faecal egg counts during naturally acquired mixed nematode infections in Moghani sheep. Annals Biol. Res. 2: 170-174.

Martins, A. C., P. L. F. Bergamasco, G. Felippelli, J. H. Tebaldi, M. F. D. Moraes, A. J. P. Testi, I. M. Lapera, & E. G. L. Hoppe. 2017. Haemonchus contortus resistence to monepantel in sheep: fecal egg count reduction tests and randomized controlled trials. Semina: Ciencias Agr. 38: 231-238.

Naumann, H. D., J. P. Muir, B. D. Lambert, L. O. Tedeschi, & M. M. Kothmann. 2013. Condensed tannins in the ruminant environment: a perspective on biological activity. J. Agr. Sci. 1: 8-20.

Nawaz, M., S. M. Sajid, M. Zubair, J. Hussain, Z. Abbasi, A. Mohi-Ud-Din, & M. Waqas. 2014. In vitro and In vivo anthelmintic activity of leaves of Azadirachta indica, Dalbergia sisso and Morus alba against Haemonchus contortus. Global Vet. 13: 996- 1001.

Odhong, C., R. G. Wahome, M. Vaarst, S. Nalubwama, M. Kiggundu, N. Halberg, & S. Githigia. 2014. In vitro anthelmintic effects of crude aqueous extracts of Tephrosia vogelii, Tephrosia villosa, and Carica papaya leaves and seeds. Afr. J. Biotechnol. 13: 4667-4672.

Paolini, V., J. P. Bergeaud, C. Grisez, F. Prevot, Ph. Dorchies, & H. Hoste. 2003. Effect of condensed tannins on goats experimentally infected with Haemonchus contortus. Vet. Parasitol. 113: 253-261.

Pathak, A. K., N. Dutta, P. S. Banerjee, T. K. Goswami, & K. Sharma. 2016. Effect of condensed tannins supplementation through leaf meal mixture on voluntary feed intake, immune response and worm. J. Parasit Dis. 40: 100-105.

Pathak, A. K. & S. P. Tiwari. 2013. Effect of high plane of nutrition on the performance of Haemonchus contortus infected kids. Vet. World. 6: 22-26.

Rezansoff, A. M., R. Laing, & J. S. Gilleard. 2016. Evidence from two independent backcross experiments supports genetic linkage of microsatellite Hcms8a20, but not other candidate loci, to a major ivermectin resistance locus in Haemonchus contortus. Intl. J. Parasitol. 46: 653-661.

Subarpriya, R. & S. Nagini. 2005. Medicinal Properties of Neem Leaves: A Review. Curr. Med. Chem. 5:149-156.

Tresia, G. E., D. Evvyernie, & R. Tiuria. 2016. Phytochemical Screening and in Vitro Ovicidal, Larvacidal, and Nematicidal Effects of Murraya paniculata (L.) Jack Extract on Gastrointestinal Parasites of Goats. Med. Pet. 39: 173-179.

Vieira, T. M., L. D. Fonseca, G. A. Bastos, V. O. Vasconcelos, M. L. F. Silva, F. Morais-Costa, A. V. P. Ferreira, N. J. F. Oliveira, & E. R. Duarte. 2017. Control of Haemonchus contortusi in sheep using basidiocarps of Agaricus blazei Murril. Vet. Res. Commun. 41: 99-106.



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