Les Tanins et les Flavonoïdes dans l'Activité Anthelminthique in Vivo et in Vitro de Newbouldia Laevis et Zanthoxylum Zanthoxyloïdes sur les Vers Adultes de Haemonchus Contortus

  • Erick Virgile Bertrand Azando Laboratoire d’Ecologie, de Santé et de Production Animales (LESPA), Faculté d’Agronomie (FA), Université de Parakou (UP), Parakou, Bénin Laboratoire de Biotechnologies et d’Amélioration Animale (LABAA), Institut des Sciences Biomédicales Appliquées (ISBA), Cotonou, Bénin Laboratoire d’Ethnopharmacologie et de Santé Animale (LESA), Faculté des Sciences Agronomiques (FSA), Université d’Abomey-Calavi (UAC), Recette Principale, Cotonou, Bénin
  • Pascal Abiodoun Olounladé Laboratoire des Sciences Animale et Halieutique (LaSAH), Unité de Recherche en Zootechnie et Système d’Élevage (URZoSE), Ecole de Gestion et d’Exploitation des Systèmes d’Elevage (EGESE), Université Nationale d’Agriculture (UNA), Porto-Novo, Bénin Laboratoire de Biotechnologies et d’Amélioration Animale (LABAA), Institut des Sciences Biomédicales Appliquées (ISBA), Cotonou, Bénin Laboratoire d’Ethnopharmacologie et de Santé Animale (LESA), Faculté des Sciences Agronomiques (FSA), Université d’Abomey-Calavi (UAC), Recette Principale, Cotonou, Bénin
  • Mawulé Sylvie Hounzangbé-Adoté Laboratoire d’Ethnopharmacologie et de Santé Animale (LESA), Faculté des Sciences Agronomiques (FSA), Université d’Abomey-Calavi (UAC), Recette Principale, Cotonou, Bénin
Keywords: Zanthoxylum zanthoxyloïdes, Newbouldia laevis, Haemonchus contortus, tanins, PVPP, PEG

Abstract

Aux fins de déterminer in vitro et in vivo la part des tanins et des flavonoïdes dans l’activité anthelminthique de Newbouldia laevis et Zanthoxylum zanthoxyloïdes  (Fagara zanthoxyloïdes) sur les vers adultes de  H. contortus, des ovins Djallonké infestés par H. contortus ont été traités ad libitum avec des feuilles fraiches de  Z. zanthoxyloïdes associées ou non à un inhibiteur de tanins, le poly éthylène glycol (PEG) sur 7 jours consécutifs. Des vers adultes de H. contortus ont été également mis en contact d’extraits de N. laevis et Z. zanthoxyloïdes en présence ou non de polyvinyle polypyrollidone (PVPP) un autre inhibiteur de tanins. In vivo, le Fagara a réduit de 20% l’excrétion fécale et l’inhibition de l’excrétion  par le PEG n’est pas significative. Le nombre de vers adultes d’H. contortus a été réduit de 39%. Le traitement par les feuilles de Fagara a significativement réduit (p < 0,05) le nombre d’œuf par ver femelle d’H. contortus et cette réduction est inhibée par le PEG. In vitro, les vers adultes de H. contortus ont été très sensibles à l’effet des extraits dans les premières 24h et au-delà, où les différences d'effet avec le contrôle sont très significatives pour le N. laevis notamment la fraction cyclohexanique (p < 0,001) et significatives pour le Fagara avec la fraction acétonique (p < 0,04). Le PVPP a partiellement restauré la mobilité jusqu'à 36h pour N. laevis respectivement 25% et 40% pour les extraits acétonique et cyclohexanique. Quant au Fagara le PVPP a eu une action modeste sur l'extrait acétonique (33% de restauration en 24h) mais a mieux inhibé l’effet de l’extrait cyclohexanique (100% en 24h et 40% en 36h).

 

In order to determine in vitro and in vivo the involvement of tannins and flavonoids in the anthelmintic activity of Newbouldia laevis and Zanthoxylum zanthoxyloïdes (Fagara zanthoxyloïdes) on adult worms of H. contortus, Djallonke ovine infested by H. contortus have been treated ad libitum with cool leaves of Z. zanthoxyloïdes associated or no to an inhibitor of tannins, the poly ethylene glycol (PEG) on seven consecutive days. Adult worms of H. contortus have also been put in contact of extracts of N. laevis and Z. zanthoxyloïdes in presence or no of polyvinyle polypyrollidone (PVPP) another inhibitor of tannins. In vivo, the Fagara decreased by 20% the fecal excretion and the inhibition of the excretion by the PEG is not significative. The number of adult worms of H. contortus has been decreased by 39%. The treatment by the leaves of Fagara reduced meaningfully (p < 0.05) the number of .eggs by female worm of H. contortus and this reduction is inhibited by the PEG. In vitro, adult worms of H. contortus was very sensitive to the effect of the extracts in the first 24h and beyond, where the differences of effect with the control are very meaningful for the N. laevis notably the cyclohexanic fraction (p < 0.01) and meaningful for the Fagara with the acetonic fraction (p < 0.04). The PVPP partially restored the mobility until 36h for N. laevis respectively 25% and 40% for the acetonic and cyclohexanic extracts. With regard to the Fagara the PVPP had a modest action on acetonic extract (33% of restoration in 24h) but inhibited the effect of cyclohexanic extract (100% in 24h and 40% in 36h) better.

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References

1. Akouedegni C.G., Daga F.D., Olounlade P.A., Allowanou G.O., Ahoussi E., Tamboura Hamidou H., Hounzangbe-Adote M.S. (2019). Evaluation in vitro et in vivo des propriétés anthelminthiques de feuilles de Spondias mombin sur Haemonchus contortus des ovins Djallonké. Agronomie Africaine, 31(2): 213-222.
2. Alawa CBI., Adamu AM., Gefu JO., Ajanusi OJ., Abdu PA., Chiezey NP., Alawa JN., Bowman DD. (2003). In vitro screening of two Nigerian medicinal plants (Vernonia amygdalina and Annona senegaiensis) for anthelmintic activity, Vet. Parasitol. 113, 73 – 81.
3. Athanasiadou, S., Kyriazakis, I., Jackson, F., Coop, R.L. (2000). Consequences of long-term feeding with condensed tannins on sheep parasitised with Trichostrongylus colubriformis. Int. J. Parasitol. 30 (9), 1025-1033.
4. Athanasiadou, S., Kyriazakis, I., Jackson, F., Coop, R.L. (2001). Direct anthelmintic effects of condensed tannins towards different gastrointestinal nematodes of sheep: in vitro and in vivo studies. Vet. Parasitol. 99 (3), 205-219.
5. Athanasiadou, S., Kyriazakis, I. (2004). Plant secondary metabolites: antiparasitic effects and their role in ruminant production systems. Proc. Nutr. Soc. 63, 631–639.
6. Azando, E.V.B., Hounzangbé–Adoté, M.S., Olounladé, P.A., Brunet, S., Fabre, N., Valentin, A., Hoste. H. (2011). Involvement of tannins and flavonoids in the in vitro effects of Newbouldia laevis and Zanthoxylum zanthoxyloïdes extracts on the exsheathment of third-stage infective larvae of gastrointestinal nematodes. Vet. Parasitol. 180, 292-297.
7. Azando E.V.B., Olounladé A.P., Hounzangbé-Adoté M.S., Tam Ha T.B., Fabre N., Valentin A. (2017). Contrôle des parasitoses gastro-intestinales ovines par l’huile essentielle de Zanthoxylum zanthoxyloïdes (Fagara zanthoxyloïdes). Revue de Médecine Vétérinaire. 168, 10-12, 205-212.
8. Barrau, E., Fabre, N., Fouraste, I. et Hoste, H. (2005). Effect of bioactive compounds from Sainfoin (Onobrychis viciifolia Scop.) on the in vitro larval migration of Haemonchus contortus: role of tannins and flavonol glycosides. Parasitology 131, 531-538.
9. Beugnet, F., Kerboeuf, D. (1997). La résistance aux antiparasitaires chez les parasites des ruminants. Le Point Vétérinaire, 28, n° spécial « Parasitologie des ruminants », 1949-1956.
10. Brunet, S., Aufrere, J., El Babili, F., Fouraste, I., Hoste, H., (2007). The kinetics of exsheathment of infective nematode larvae is disturbed in the presence of a tannin-rich plant extract (sainfoin) both in vitro and in vivo. Parasitology 134, 1253–1262.
11. Cenci, F.B., Louvandini, H., Mcmanus, C.M., Dell'porto, A., Costa, D.M., Araujo, S.C., Minho, A.P., Abdalla, A.L. (2007). Effects of condensed tannin from Acacia mearnsii on sheep infected naturally with gastrointestinal helminthes. Vet. Parasitol. 144 (1-2), 132-137.
12. Chartier, C., Soubirac, F., Pors, I., Silvestre, A., Hubert, J., Couquet, C., Cabaret, J. (2001). Prevalence of anthelmintic resistance in gastrointestinal nematodes of dairy goats under extensive management conditions in southwestern France. J. Helminthol. 75, 325-330.Couillerot, E,. Caron, L., Audran, J.C., Molinatti, P., Le Men Olivier, L., Jardillet, J.C., Chenieux, J.C. (1994). Benzophenanthridine and furoquinoline accumulation in cell suspension culture of Fagara zanthoxyloïdes. Phytochemistry 37, 425-428.
13. Diéguez-Hurtado, R., Garrido, G., Prieto Gonzalez, S. Iznaga, Y., Gonzalez, L., Molina Tores, J., Curini, M., Epifano, F., Marcotullio, M.C. (2003). Antfungal activity of some Cuban Zanthoxylum species. Fitoterapia 74, 384-386.
14. Enwerem NM., Okogun JI., Wambebe CO., Okorie DA., Akah PA. (2001). Anthelmintic activity of the stem bark extracts of Berlina grandiflora and one of its active principles, Betulinic acid. Phytomed , 8: 112 -114.
15. Eyong, K.O., Krohn, K., Hussain, H., Folefoc, G.N., Nkengfack, A.E.,,Schulz, B., Hu, Q. (2005). Newbouldiaquinone and newbouldiamide: a new naphthoquinone-anthraquinone coupled pigment and a new ceramide from Newbouldia laevis. Chem. Pharm. Bull. (Tokyo) 53, 616–619.
16. Eyong, K.O., Folefoe, G.N., Kuete, V., Beng, V.P., Krohn, K., Hussain, H., Nkengfack, A.E., Saeftel, M., Sarite, S.R., Hoerauf, A. (2006). New446 bouldiaquinone A: a naphtoquinone–anthraquinone ether coupled pigments, as a potential antimicrobial and antimalarial agents for Newbouldia laevis. Phytochemistry 67, 605–609.
17. Foo, L.Y., Newman, R., Waghorn, G., Mc Nabb, W.C., Ulyatt, M.J. (1996). Proanthocyanidins from Lotus corniculatus. Phytochemistry 41, 617-624.
18. Foo, L.Y., Lu, Y., Mc Nabb, W.C., Waghorn, G., Ulyatt, M.J. (1997). Proanthocyanidins from Lotus pedunculatus. Phytochemistry 45, 1689-1696.
19. Foo, L.Y., Lu, Y., Molan, A.L., Woodfield, D.R., Mc Nabb, W.C. (2000). The phenols and prodelphinidins of white clover flowers. Phytochemistry 54 (5), 539-548.
20. Githiori, J.B., Athanasiadou, S., Thamsborg, S.M. (2006). Use of plants in novel approaches for control of gastrointestinal helminths in livestock with emphasis on small ruminants. Vet. Parasitol. 139 (4), 308-320.
21. Hagerman, A.E., (1987). Radial diffusion method for determining tannin in plant extracts. J. Chem. Ecol. 13, 437–449.
22. Hansen J., Perry B. (1995). Épidémiologie, diagnostic et prophylaxie des helminthiases des ruminants domestiques, FAO-Rome, Italie, 7ème édition, 176 p.
23. Heckendorn, F., Häring, D.A., Maurer, V., Zinsstag, J., Langhans, W., Hertzberg, H. (2006). Effect of sainfoin (Onobrychis viciifolia) silage and hay on established populations of Haemonchus contortus and Cooperia curticei in lambs. Vet. Parasitol. 142 (3-4), 293-300.
24. Heckendorn, F., Häring, D.A., Maurer, V., Senn, M., Hertzberg, H. (2007). Individual administration of three tanniferous forage plants to lambs artificially infected with Haemonchus contortus and Cooperia curticei. Vet. Parasitol. 146, 123–134.
25. Hoste, H., Jackson, F., Athanasiadou, S., Thamsborg, S.M., Hoskin, S.O. (2006). The effects of tannin-rich plants on parasitic nematodes in ruminants.Trends Parasitol. 22, 253–261.
26. Hoste, H., Gaillard, H., Le Frileux, Y. (2005). Consequences of the regular distribution of sainfoin hay on gastrointestinal parasitism with nematodes and milk production in dairy goats. Small Rum. Res. 59 (2-3), 265-271.
27. Hoste, H., Chartier, C. (1993). Comparison of the effects on milk production of concurrent infection with Haemonchus contortus and Trichostrongylus colubriformis in high- and low-producing dairy goats. Am. J. Vet. Res. 54 (11), 1886-1893.
28. Hounzangbé-Adoté, S., Zinsou, F.E., Hounkpè, V., Moutairou, K., Hoste, H. (2005a). In vivo effects of fagara leaves on sheep infected with gastrointestinal nematodes. Trop. Anim. Health Product 37, 205–214.
29. Hounzangbé-Adoté, S., Paolini, V., Fouraste, I., Moutairou, K., Hoste, H. (2005b). In vitro effects of four tropical plants on the intestinal parasitic nematode, Haemonchus contortus. Res. Vet. Sci. 78, 155–160.
30. Kahn, L.P., Diaz-Hernandez, A. (2000). Tannins with anthelmintic properties. In: Tannins in livestock and human nutrition: ACIAR proceeding n.92, international workshop (Brooker, ed.), Adelaide.
31. Kerharo, J., Adam, J.G. (1974). La pharmacopée sénégalaise traditionnelle. Plantes médicinales et toxiques, Vigot frères, Paris, 1007 p.
32. Ketzis, J.K., Vercruysse, J., Stromberg, B.E., Larsen, M., Athanasiadou, S., Houdijk, J.G. (2006). Evaluation of efficacy expectations for novel and non-chemical helminth control strategies in ruminants. Vet. Parasitol. 139 (4), 321-335.
33. Kloosterman A., Albers G.A.A., Van Den Brink R.(1978). Genetic variations among calves in resistance to nematode parasites. Vet. Parasitol., 4, 353-368.
34. Knox, M.R., Torres-Acosta, J.F., Aguilar-Caballero, A.J. (2006). Exploiting the effect of dietary supplementation of small ruminants on resilience and resistance against gastrointestinal nematodes. Vet. Parasitol. 139 (4), 385-393.
35. Kuiseu J., Olounladé A.P., Houssoukpè G.C., Sounkéré A.B.T.T., Zinsou A.T.F Konmy B.S.B., Dansou C.D., Moudachirou I., Babatoundé S., Hounzangbé-Adoté M.S., Edorh A.P. (2022). In vitro effects of Anogeissus leiocarpus and Adansonia digitata on two life-cycle stages of Haemonchus contortus, a gastrointestinal parasite of small ruminants. Animal and Veterinary Sciences, 10(1): 1-7.
36. Lange, K.C., Olcott, D.D., Miller, J.E., Mosjidis, J.A., Terrill, T.H., Burke, J.M., Kearney, M.T. (2006). Effect of Sericea lespedeza (Lespedeza cuneata) fed as hay, on natural and experimental Haemonchus contortus infections in lambs. Vet. Parasitol. 141 (3-4), 273-278.
37. Makkar, H.P.S. (2003). Effects and fate of tannins in ruminant animals, adaptation to tannins, and strategies to overcome detrimental effects of feeding tannin-rich feeds. Small Rum. Res. 49 (3), 241-256.
38. Mara, S.P., Arruda Jao, B., Fernandes Paulo, C., Viera, M., Fatima Das, G.F., Da Silva, Jose, R.P. (1992). Chemistry of Zanthoxylum rhoifolium. A new secofuraquinoline alkaloids. Biochem. System. Ecol. 20, 173-178.
39. Marie-Magdeleine, C., Hoste, H., Mahieu, M., Varo, H., Archimede, H. (2009). In vitro effects of Cucurbita moschata seed extracts on Haemonchus contortus. Vet. Parasitol. 161, 99–105.
40. Martínez-Ortíz-de-Montellano, C., C Arroyo-López, C., Fourquaux, I., Torres-Acosta, J.F.J., Sandoval-Castro, C.A., Hoste, H. (2013). Scanning electron microscopy of Haemonchus contortus exposed to tannin-rich plants under in vivo and in vitro conditions. Experimental Parasitology. 133(3): 281-286
41. Martínez-Ortiz-de-Montellano, C.; Vargas-Magaña, J.J.; Canul-Ku, L.; Miranda-Soberanis, R.; Capetillo-Leal, C.; Sandoval-Castro, C.A.; Hoste, H.; Torres-Acosta, J.F. (2010). Effect of a tropical tannin-rich plant, Lysiloma latisiliquum on adult populations of Haemonchus contortus in sheep. Vet. Parásitol. 172, 283-290.
42. Minaflinou Sacca Sidi I.Y., Azando E.V.B., Olounladé P.A., Hounzangbé-Adoté M.S. (2015).
43. Effets combinés des feuilles de Newbouldia laevis et de Zanthoxylum zanthoxyloïdes sur les nématodes parasites gastro-intestinaux des ovins Djallonké. International Journal of Biological and Chemical Sciences. 9 (4): 2078-2090.
44. Molan, A.L., Hoskin, S.O., Barry, T.N., Mc Nabb, W.C. (2000). Effect of condensed tannins extracted from four forages on the viability of the larvae of deer lungworms and gastrointestinal nematodes. Vet. Rec. 147 (2), 44-48.
45. Niezen, J.H., Charleston, W.A.G., Robertson, H.A., Shelton, I.D., Waghorn, G., Green, R. (2002). The effect of feeding sulla (Hedysarum coronarium) or lucerne (Medicago sativa) on lamb parasite burdens and development of immunity to gastrointestinal nematodes. Vet. Parasitol. 105, 229-245.
46. Olounladé P.A., Azando E.V.B., Attakpa E.Y., Gbnéou J.D., Alowanou G.G, Tchétan E., Dansou C.C., Hounzangbé-Adoté M.S., Gbaguidi F., Moudachirou M., Hoste H., Valentin A. (2017a). In vitro study on the role of the tannins of Newbouldia laevis and Zanthoxylum zanthoxyloïdes on infective larvae of Trichostrongylus colubriformis. African Journal of
47. Agricultural Research. Vol. 12(50): 3513-3519.
48. Olounladé A.P., Attakpa Y.E., Azando E.V.B., Hounzangbé-Adoté M. S., Hoste H. (2017b). Effet in vivo de Newbouldia laevis (Bignoniaceae) sur des strongles gastro-intestinaux des moutons. European Scientific Journal. 13 (12) : 335-351.
49. Olounladé P.A., Azando E. V. B., Gbaguidi F., Gbénou J., Hoste H., Valentin A., Moudachirou M., Hounzangbé-Adoté M. S. (2018). Variability of chemical composition of Newbouldia laevis and Zanthoxylum zanthoxyloïdes related to environmental factors. Annales des sciences agronomiques 22(1) : 73-92.
50. Paolini, V., Bergeaud, J.P., Grisez, C., Prevot, F., Dorchies, P., Hoste, H. (2003a). Effects of condensed tannins on goats experimentally infected with Haemonchus contortus. Vet. Parasitol. 113 (3-4), 253-261.
51. Paolini, V., Dorchies, P., Hoste, H. (2003b). Effects of sainfoin hay on gastrointestinal nematode infections in goats. Vet. Rec. 152 (19), 600-601.
52. Paolini, V., Fouraste, I., Hoste, H. (2004). In vitro effects of three woody plant and sainfoin extracts on 3rd-stage larvae and adult worms of three gastrointestinal nematodes. Parasitology 129 (1), 69-77.
53. Paolini, V., De La Farge, F., Prevot, F., Dorchies, P., Hoste, H. (2005). Effects of the repeated distribution of sainfoin hay on the resistance and the resilience of goats naturally infected with gastrointestinal nematodes. Vet. Parasitol. 127 (3-4), 277-283.
54. Perrett, S., Whitfield, P.J. (1995). Atanine (3-dimethylallyl-4-methoxy-2-quinolone), an alkaloid with anthelmintic activity from the Chinese medicinal plant, Evodia rutaecarpa. Planta Medica 61, 276-278.
55. Rogosic, J., Estell, R.E., Ivankovic, S., Kezic, J., Razov, J. (2008). Potential mechanisms to increase shrub intake and performance of small ruminants in mediterranean shrubby ecosystems. Small Rum. Res. 74 (1-3), 1-15.
56. Rojas, D.K., Lopez, J., Tejada, I., Vazquez, V., Shimada, A., Sanchez, D., Ibarra, F. (2006). Impact of condensed tannins from tropical forages on Haemonchus contortus burdens in Mongolian gerbils (Meriones unguiculatus) and Pelibuey lambs. An. Feed Sci. Technol. 128, 218-228.
57. Shaik, S.A., Terrill, T.H., Miller, D., Kouakou, B., Kamman, G., Kallu, R.K., Mosjidis, J. (2004). Effects of feeding Sericea lespedeza hay to goats infected with Haemonchus contortus. South African J. Anim. Sci. 34, 234-237.
58. Shaik, S.A., Terrill, T.H., Miller, J.E., Kouakou, B., Kannan, G., Kaplan, R.M., Burke, J.M., Mosjidis, J.A. (2006). Sericea lespedeza hay as a natural deworming agent against gastrointestinal nematode infection in goats. Vet. Parasitol. 139 (1-3), 150-157.
59. Silanikove, N., Perevolotsky, A., Provenza, F.D. (2001). Use of tanninbinding chemicals to assay for tannins and their negative postingestive effects in ruminants. Anim. Feed Sci. Technol. 91 (1-2), 69-81.
60. Streit, W. et Fengel, D. (1994). Purified tannins from quebracho colorado. Phytochemistry 36, 481-484.
61. Tchetan E., Azando E.V.B., Olounladé P.A., Alowanoua G.G., Hounzangbé-Adoté S.M. (2020). In vitro effects of tannin and extracts of Bridelia ferruginea and Mitragyna inermis on the exsheathment of infective larvae of Haemonchus contortus. International Journal of Veterinary Science and Medicine, 8(1): 93–99.
62. Thamsborg, S.M., Mejer, H., Bandier, M., Larsen, M. (2003). Influence of different forages on gastrointestinal nematode infections in grazing lambs. In: The 19th International Conferences of WAAVP, pp. 189, New Orleans, USA.
63. Tringali, C., Carmela, C., Valeria Cali, V., Simmonds, M.S.J. (2001). Antifeedant constituents from Fagara macrophylla. Fitoterapia 72, 538-543.
64. Tzamaloukas, O., Athanasiadou, S., Kyriazakis, I., Jackson, F., Coop, R.L. (2005). The consequences of short-term grazing of bioactive forages on established adult and incoming larvae populations of Teladorsagia circumcincta in lambs. Int. J. Parasitol. 35 (3), 329-335.
65. Waghorn, T.S., Molan, A.L., Deighton, M., Alexander, R.A., Leathwick, D.M., Mc Nabb, W.C., Meagher, L.P. (2006). In vivo anthelmintic activity of Dorycnium rectum and grape seed extract against Ostertagia (Teladorsagia) circumcincta and Trichostrongylus colubriformis in sheep. N. Z. Vet. J. 54 (1), 21-27.
66. Waller, P.J. (1997). Anthelmintic resistance. Vet. Parasitol. 72 (3-4), 391-405; 405-412.
67. Wolstenholme, A.J., Fairweather, I., Prichard, R.K., Samson-Himmelstjerna, G., Sangster, N.C. (2004). Drug resistance in veterinary helminths. Trends Parasitol. 20 (10), 469-476.
68. Zinsou A.T.F., Olounladé A.P., Adenilé D.A., Alowanou G.G., Hounzangbé-Adoté M.S., Babayemi O.J. (2021). In vitro effects of maize silage extracts on Haemonchus contortus, gastrointestinal nematode parasite in Red Maradi Goats. Journal of Animal & Plant Sciences, 48 (2): 8686-8702.
Published
2022-11-20
How to Cite
Bertrand Azando, E. V., Olounladé, P. A., & Hounzangbé-Adoté, M. S. (2022). Les Tanins et les Flavonoïdes dans l’Activité Anthelminthique in Vivo et in Vitro de Newbouldia Laevis et Zanthoxylum Zanthoxyloïdes sur les Vers Adultes de Haemonchus Contortus. European Scientific Journal, ESJ, 11, 493. Retrieved from https://eujournal.org/index.php/esj/article/view/16133
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ESI Preprints