IN VITRO ANTHELMINTIC EFFICACY OF EXTRACTS OF CITRUS AURANTIFOLIA (CHRISTM) SWINGLE FRUIT PEELS AGAINST HELIGMOSOMOIDES BAKERI OVA AND LARVAE
In vitro ovicidal and larvicidal activities of extracts of Citrus aurantifolia fruit were investigated on the eggs and first stage larvae of Heligmosomoides bakeri. The fruit peels of Citrus aurantifolia were extracted with methanol. The crude methanol extract (CME) was dissolved in water and serially partitioned with hexane and ethylacetate to give hexane extract (HE), ethylacetate extract (EE) and aqueous methanol extract (AME). Different concentrations (0.625, 1.25, 2.5, 5, 10, and 20 mg/ml) of the extracts were prepared by dissolving the extracts in distilled water. Two hundred microlitres of each concentration was incubated with the larvae and eggs of H. bakeri contained in 0.2 ml solution in a 96-well microtitre plates and incubated at room temperature for 24 and 48 hours, respectively. Distilled water and albendazole were used as non-treated and treated controls, respectively. At 20 mg/ml, the CME, EE, HE and AME inhibited the hatching of H. bakeri eggs by 100, 98.2, 97.8 and 97.7 %, respectively. Similarly, CME, AME, EE and HE caused death of the larvae of H. bakeri by 100, 90.8, 85.9 and 83.3 %, respectively. All the extracts showed significant (***p<0.001) anthelmintic effect when compared with the non-treated (DW) control. This study demostrates that Citrus aurantifolia fruit peels possess anthelmintic activity that might be caused by one or more of the secondary metabolites contained in the plant.
2. Waller PJ. Nematode parasite control of livestock in the tropics/subtropics: the need for novel approaches. Int J Parasitol 1997;27:1193-201.
3. Githiori JB, Hoglund J, Waller PJ, Baker RL. Evaluation of anthelmintic properties of some plants used as livestock dewormers against Haemonchus contortus infections in sheep. Parasitol 2004;129:245â€“53.
4. Saeed M, Iqbal Z, Jabbar A. Oxfendazole resistance in gastrointestinal nematodes of beetal goats at livestock farms of Punjab (Pakistan). Acta Vet Brno 2007;76:79â€“85.
5. Gasbarre CL, Stout LW, Leighton AE. Gastrointestinal nematodes of cattle in the northeastern US: results of a producer survey. Vet Parasitol 2001;101:29â€“44.
6. Kudi AC, Myint SH. Antiviral activity of some Nigerian medicinal plant extracts. J Ethnopham 1999;68:289-94.
7. Abo KA, Fred-Jaiyessimi AA, Jaiyesimi AEA. Ethnobotanical studies of medicinal plants used in the management of diabetes mellitus in South Western Nigeria. J Ethnopham 2008;115:67-71.
8. Reddy JL, Reshma DH, Beena J, Spandana G. Evaluation of antibacterial and antioxidant activities of the leaf essential oil and leaf extracts of Citrus aurantifolia. Asian J Biochem Pharm Res 2012;2(2):2249â€“4626.
9. Souza A, Lamidi M, Ibrahim B, Aworet SRR, Boukandou M, Mâ€™Batchi B. Antihypertensive effect of an aqueous extract of Citrus aurantifolia (Rutaceae) (Christm.) Swingle, on the arterial blood pressure of mammal. Int Res Pharm Pharm 2011;1(7):142-8.
10. Jaiprakash RP, Jayaprakasha GK, Mahadev BC, Bhimanagouda SP. Bioactive compounds from maxican lime (Citrus aurantifolia) juice induce apoptosis in human pancreatic cells. J Agric Food Chem 2009;57(22):10933-42.
11. Nallely E, Sandoval M, Abraham G, Elizondo-TreviÃ±o E, Garza-GonzÃ¡lez E, Alvarez, et al. Chemical composition of hexane extract of Citrus aurantifolia and anti-mycobacterium tuberculosis activity of some of its constituents. Molecules 2012;17:11173-84.
12. Dongmo PMD, Tatsadjieu LN, Tchinda ES, Kuate J, Amvam PHZ, Menut C. Essential oils of Citrus aurantifolia from Cameroon and their antifungal activity against Phaeoramularia angolensis. Afr J Agric Res 2009;4(4):354-8.
13. Loizzo MR, Tundis R, Bonesi M, Menichini F, De Luca D, Colica C, et al. Evaluation of Citrus aurantifolia peel and leaves extracts for their chemical composition, antioxidant and anti-cholinesterase activities. J Sci Food Agric 2012;24(12):18930-7.
14. Wabo PJ, Payne VK, Mbogning TG, Komtangi MC, Yondo J, Ngangout MA, et al. In vitro anthelminthic efficacy of Dichrocephala integrifolia (Asteraceae) extracts on the gastro-intestinal nematode parasite of mice: Heligmosomoides bakeri (Nematoda, Heligmosomatidae). Asian Pac J Trop Biomed 2013;3(2):100-4.
15. Behnke JM, Menge DM, Noyes H. Heligmosomoides bakeri: a model for exploring the biology and genetics of resistance to chronic gastrointestinal nematode infections. Parasitol 2009;136(12):1565-80.
16. Handa SS, Khanuja SPS, Longo G, Rakesh DD. Extraction technologies for medicinal and aromatic plants. Int centre for science and high technology, Trieste; 2008. p. 21-5.
17. Trease and Evans, Pharmacognosy. Springer publication; 2007. p. 153-7.
18. Wabo PonÃ© J, Bilong BCF, Mpoame M. In vitro nematicidal activity of extracts of Canthium mannii (Rubiaceae) on different life cycle stages of Heligmosomoides polygyrus (Nematoda, Heligmosomatidae). J Helmenthol 2010;84:156-65.
19. Wabo PonÃ© J, Mpoame M, Bilong BCF, Kerboeuf D. In vitro comparative study of the nematicidal activity of ethanolic extract of the powder of the stem bark of Canthium mannii (Rubiaceae) and the MeÂ²bendazole. Revue MÃ©d VÃ©t 2005;156(12):633-6365.
20. Mali RG, Mehta AA. A review of Anthelmintic plants. Nat Prod Rad 2007;7(5):466-75.
21. Ademola O, Eloff JN. Anthelmintic efficacy of cashew (Anarcadium occidentale L.) on in vitro susceptibility of the ova and larvae of Haemonchus contortus. Afr J Biotech 2011;10(47):9700-5.
22. Kollins EN, Wabo PJ, Payne VK, Jeannette Y, Marie-Claire K. In vitro comparative effect of aqueous (Cold and Hot Water) and ethanolic extracts of the seeds of Aframomum Danielli (Zingiberaceae) on three life cycle stages of the parasitic nematode Heligmosomoides bakeri (Nematoda; Heligmosomatidae), Parasite of the Laboratory Mice (Mus Musculus). Med Aromat Plants 2012;1:11.
23. Rajarajan AT, Vijayasree VG, Kenichi W, Vijaya S, Narasimman G, Sadish SK. Anthelmintic and antimicrobial properties of fruit peels of Citrus sinensis. Pharmacologyonline 2009;1:363-8.
24. Bairagi GB, Kabra AO, Mandade RA. Anthelmintic activity of Citrus limon L. (burm) fruit peels extract in Indian adult earthworm. Int J Pharm Tech 2011;3(2):664-7.
25. Cock IE. Problems of Reproducibility and Efficacy of bioassays using crude extracts, with reference to Aloe vera. Pharmacogn Commun 2011;1(1):52-62.
26. Wabo PJ, Yondo J, Fossi TO, Marie CK, Bilong BCF, Mpoame M. The in vitro effects of Chenopodium ambrosioides (Chenopodiaceae) extracts on the parasitic nematode Heligmosomoides bakeri (Nematoda, Heligmosomatidae). J Pharmacogn Phytochem 2011;3(4):56-62.
27. Payne VK, Kollins EN, Wabo PJ, Yondo J, Komtangi M, Mpoame M, et al. In vitro comparative effect of extracts of the seeds of Embelia rowlandii (Myrsinaceae) on the eggs and L1 and L2 larval stages of the parasitic nematode Heligmosomoides bakeri (Nematoda; Heligmosomatidae). Afr J Biotech 2013;12(2):205-11.
28. Abdelqader A, Bassam Q, Diya A, GÃ¼rbÃ¼z D. Anthelmintic effects of citrus fruit peels ethanolic extracts against Ascaridia galli. Vet Parasitol 2012:1-7.
29. Pathan RK, Papi RG, Parveen P, Tananki G, Soujanya P. In vitro antimicrobial activity of citrus aurantifolia and its phytochemical screening. Asian Pac J Trop Dis 2012;2012:328-31.
30. Suleiman MM, Mamman M, Aliu YO, Ajanusi JO. Anthelmintic activity of the crude methanol extract of Xylopia aethiopica against Nippostrongylus brasiliensis in rats. Veterinarski Arhiv 2005;75:487-95.
31. Athanasiadou S, Githiori J, Kyriazakis I. Medicinal plants for helminth parasite control: facts and fiction. Animal 2007;1:1392â€“400.
32. Ngongeh LA, Fakae BB. Effect of quebracho tannin drenching on Heligmosomoides bakeri infection in mice. Int J Sci Nat 2011;2(1):132-5.
33. Bate-Smith EC. The phenolic constituents of plants and their taxonomic significance. J Linn Soc London Bot 1962;58:95-173.
34. Duncan RA. An evaluation of suitable bioassays to measure the potential of condensed tannins to inhibit the viability of sheep nematode parasites. Bachelor of Rural Science, Honours thesis, Animal Science, University of New England, Armidale, NSW, Australia; 1996. p. 34.
35. Price KR, Johnson INT, Fenwick GR. The chemistry and biological significance of saponin in food and feed stuffs. Res Food Sci 1987;26:27-135.
36. Squires JM, Foster J, Lindsay D, Caudell D, Zajac A. Efficacy of an orange oil emulsion as an anthelmintic against Haemonchus contortus in gerbils (Meriones unguiculatus) and in sheep. Vet Parasitol 2010;172:95â€“9.
37. Jimenez-Cisneros BE. Helminth ova control in waste water and sludge for Agricultural reuse. In: Grabow WOK, Editor. Encyclopedia of Life Support Systems (EOLSS); 2007. p. 3-9.
38. Riviere RE, Papich MG. Veterinary Pharmacology and Therapeutics. 9th ed. Wiley-Black; 2009. p. 1053-145