Skip to main content Accessibility help

Effects of different extracts of three Annona species on egg-hatching processes of Haemonchus contortus

  • G.S. Castañeda-Ramírez (a1) (a2), J.F.J. Torres-Acosta (a1), P. Mendoza-de-Gives (a2), J. Tun-Garrido (a1), J.A. Rosado-Aguilar (a1), J.I. Chan-Pérez (a1), G.I. Hernández-Bolio (a3), J. Ventura-Cordero (a1) (a3), K.Y. Acosta-Viana (a4) and M. Jímenez-Coello (a4)...


This study assessed the in vitro anthelmintic (AH) activity of methanol and acetone:water leaf extracts from Annona squamosa, A. muricata and A. reticulata against Haemonchus contortus eggs. The egg hatch test was used to determine the effective concentrations required to inhibit 50% of eggs hatching (EC50). The role of polyphenols on AH activity was measured through bioassays with and without polyvinylpolypyrrolidone (PVPP). Methanolic extracts mainly caused the death of eggs at the morula stage (ovicidal activity). Meanwhile, acetone:water extracts caused egg-hatching failure of developed larvae (larvae failing eclosion (LFE) activity). The lowest EC50 values against H. contortus eggs were observed for the methanolic extracts from A. reticulata and A. muricata (274.2 and 382.9 µg/ml, respectively). From the six extracts evaluated, the methanolic extracts of A. muricata, A. reticulata and A. squamosa showed the highest ovicidal activity, resulting in 98.9%, 92.8% and 95.1% egg mortality, respectively. When the methanolic extract of A. squamosa was incubated with PVPP, its AH activity increased. Similarly, when acetone:water extracts of A. muriata and A. reticulata were incubated with PVPP, their LFE activity increased. Alkaloids were only evident in methanolic extracts, irrespective of PVPP incubation. The presence of acetogenins was not observed. In conclusion, methanolic extracts obtained from leaves of A. muricata, A. reticulata and A. squamosa showed ovicidal activity affecting the morula of H. contortus eggs, with minor LFE activity. Meanwhile, acetone:water extracts showed mostly LFE activity, with a lower proportion of ovicidal activity.


Corresponding author

Author for correspondence: P. Mendoza-de-Gives, E-mail:


Hide All
Alonso-Díaz, MA, Torres-Acosta, JFJ, Sandoval-Castro, CA, Aguilar-Caballero, AJ and Hoste, H (2008) In vitro larval migration and kinetics of exsheathment of Haemonchus contortus larvae exposed to four tropical tanniniferous plant extracts. Veterinary Parasitology 153, 313319.
Arceo-Medina, GN, Rosado-Aguilar, JA, Rodríguez-Vivas, RI and Borges-Argaez, R (2016) Synergistic action of fatty acids, sulphides and stilbene against acaricide-resistant Rhipicephalus microplus ticks. Veterinary Parasitology 228, 121125.
Badri, VD, Zolla, G, Bakker, GM, Manter, KD and Vivanco, MJ (2013) Potential impact of soil microbiomes on the leaf metabolome and on herbivore feeding behavior. New Phytologist 198, 264273.
Castañeda-Ramírez, GS, Torres-Acosta, JFJ, Sandoval-Castro, CA, González-Pech, PG, Parra-Tabla, VP and Mathieu, C (2017) Is there a negative association between the content of condensed tannins, total phenols, and total tannins of tropical plant extracts and in vitro anthelmintic activity against Haemonchus contortus eggs? Parasitology Research 116, 33413348.
Castañeda-Ramírez, GS, Rodríguez-Labastida, M, Ortiz-Ocampo, GI, González-Pech, PG, Ventura-Cordero, J, Borges-Argáez, R, Torres-Acosta, JFJ, Sandoval-Castro, CA and Mathieu, C (2018) Nutraceutical potential of ten plant species from tropical deciduous forest: in vitro nutritional value and anthelmintic activity against Haemonchus contortus. Parasitology Research 117, 39793991.
Castañeda-Ramírez, GS, Rodríguez-Labastida, M, Ortiz-Ocampo, GI, González-Pech, PG, Ventura-Cordero, J, Borges-Argáez, R, Torres-Acosta, JFJ, Sandoval-Castro, CA and Mathieu, C (2018) Nutraceutical potential of ten plant species from tropical deciduous forest: In vitro nutritional value and anthelmintic activity against Haemonchus contortus. Parasitology Research 117, 39793991.
Castillo-Mitre, GF, Olmedo-Juárez, A, Rojo-Rubio, R, et al. (2017) Caffeoyl and coumaroyl derivatives from Acacia cochiacantha exhibit ovicidal activity against Haemonchus contortus. Journal of Ethnopharmacology 204, 125131.
Chan-Pérez, JI, Torres-Acosta, JFJ, Sandoval-Castro, AC, Hoste, H, Castañeda-Ramírez, GS, Vilarem, G and Mathieu, C (2016) In vitro susceptibility of ten Haemonchus contortus isolates from different geographical origins towards acetone:water extracts of two tannin rich plants. Veterinary Parasitology 217, 5360.
Coles, G, Bauer, C, Borgsteede, F, Klei, T, Taylor, M and Waller, P (1992) Methods for the detection of anthelmintic resistance in nematodes of veterinary importance. Veterinary Parasitology 44, 3544.
Coria-Téllez, AV, Montalvo-Gónzalez, E, Yahia, EM and Obledo-Vázquez, EN (2018) Annona muricata: a comprehensive review on its traditional medicinal uses, phytochemicals, pharmacological activities, mechanisms of action and toxicity. Arabian Journal of Chemistry 11, 662691.
Cortes-Morales, JA, Olmedo-Juárez, A, Trejo-Tapia, G, González-Cortazar, M, Domínguez-Mendoza, BE, Mendoza-de Gives, P and Zamilpa, A (2019) In vitro ovicidal activity of Baccharis conferta Kunth against Haemonchus contortus. Experimental Parasitology 197, 2028.
Ferreira, LE, Castro, PMN, Chagas, ACS, França, SC and Beleboni, RO (2013) In vitro anthelmintic activity of aqueous leaf extract of Annona muricata L. (Annonaceae) against Haemonchus contortus from sheep. Experimental Parasitology 134, 327332.
García-Hernández, C, Rojo-Rubio, R, Olmedo-Juárez, A, et al. (2019) Galloyl derivatives from Caesalpinia coriaria exhibit in vitro ovicidal activity against cattle gastrointestinal parasitic nematodes. Experimental Parasitology 200, 1623.
Hernández-Bolio, GI, Kutzner, E, Eisenreich, W, Torres-Acosta, JFJ and Peña-Rodríguez, LM (2018) The use of 1 H-NMR metabolomics to optimise the extraction and preliminary identification of anthelmintic products from the leaves of Lysiloma latisiliquum. Phytochemical Analysis 29, 413420.
Hoste, H, Torres-Acosta, JFJ, Sandoval-Castro, CA, Mueller-Harvey, I, Sotiraki, S, Louvandini, H, Thamsborg, SM and Terrill, TH (2015) Tannin containing legumes as a model for nutraceuticals against digestive parasites in livestock. Veterinary Parasitology 212, 517.
Hoste, H, Torres-Acosta, JFJ, Quijada, J, Chan-Pérez, I, Dakheel, MM, Kommuru, DS, Mueller-Harvey, I and Terrill, TH (2016) Interactions between nutrition and infections with Haemonchus contortus and related gastrointestinal nematodes in small ruminants. pp. 239351 in Gasser, RB and von Samson-Himmelstjerna, G (Eds) Haemonchus contortus and Haemonchosis – past, present and future trends. Advances in Parasitology. New York, Academic Press.
Jackson, F and Hoste, H (2010) In vitro methods for the primary screening of plant products for direct activity against ruminant gastroitestinal nematodes. pp. 2545 in Vercoe, PE, Makkar, HPS and Schlink, AC (Eds) In vitro screening of plant resource for extra-nutritional attributes in ruminants: nuclear and related methodologies. Dordrecht, Springer Science+Business Media.
Jasso-Díaz, G, Hernández, GT, Zamilpa, A, Becerril-Pérez, CM, Ramírez-Bribiesca, JE, Mernández-Mendo, O, Sánchez-Arrollo, H, González-Cortazar, M and Mendoza-de Gives, P (2017) In vitro assessment of Argemone Mexicana, Taraxacum officinale, Ruta chalepensis and Tagetes filifolia against Haemonchus contortus eggs and infective (L3) larvae. Microbial Pathogenesis 109, 162168.
Jork, H (1990) Thin-layer chromatography reagents and detection Methods. 3rd edn. Wemhem, Federal Republic of Germany, Verlagsgesellschaft mbH VCH. p. 496.
Kamaraj, C and Rahuman, AA (2011) Efficacy of anthelmintic properties of medicinal plant extracts against Haemonchus contortus. Research in Veterinary Science 91, 400404.
Kamaraj, C, Rahuman, AA, Elango, G, Bagavan, A and Zahir, AA (2011) Anthelmintic activity of botanical extracts against sheep gastrointestinal nematodes, Haemonchus contortus. Parasitology Research 109, 3745.
Lacey, E (1988) The role of the cytoskeletal protein, tubulin, in the mode of action and mechanism of drug resistance to benzimidazoles. International Journal for Parasitology 18, 885936.
LeOra Software (2004) Polo Plus. Probit and logit analysis. Berkeley, CA, LeOra Software.
Makkar, HP, Blümmel, M and Becker, K (1995) Formation of complexes between polyvinyl pyrrolidones or polyethylene glycols and tannins, and their implication in gas production and true digestibility in in vitro techniques. British Journal of Nutrition 73, 897913.
Prichard, RK (1973) The fumarate reductase reaction of Haemonchus contortus and mode of action of some anthelminthics. International Journal for Parasitology 3, 409417.
Rosado-Aguilar, JA, Aguilar-Caballero, AJ, Rodríguez-Vivas, RI, Borges-Argaez, R, García-Vázquez, Z and Méndez-González, M (2010) Acaricidal activity of extracts from Petiveria alliacea (Phytolaccaceae) against the cattle tick, Rhipicephalus (Boophilus) microplus (Acari: ixodidae). Veterinary Parasitology 168, 299303.
Sandoval-Castro, CA, Torres-Acosta, JFJ, Hoste, H, Salem, AZM and Chan-Pérez, JI (2012) Using plant bioactive materials to control gastrointestinal tract helminths in livestock. Animal Feed Science and Technology 176, 192201.
Souza, M, Belvi, C, Morais, SM, Costa, C, Silva, A and Braz-filho, R (2008) Anthelmintic acetogenin from Annona squamosa L. Seeds. Anais da Academia Brasileira de Ciências 80, 271277.
Statpoint Technologies Inc. (2005) Statgraphics Centurion XV. Warranton, VA, Statpoint Technologies Inc.
Vargas-Magaña, JJ, Torres-Acosta, JFJ, Aguilar-Caballero, AJ, Sandoval-Castro, CA, Hoste, H and Chan-Pérez, JI (2014) Anthelmintic activity of acetone-water extracts against Haemonchus contortus eggs: interactions between tannins and other plant secondary compounds. Veterinary Parasitology 206, 322327.
von Samson-Himmelstjerna, G, Coles, GC, Jackson, F, et al. (2009) Standardization of the egg hatch test for the detection of benzimidazole resistance in parasitic nematodes. Parasitology Research 105, 825834.
Yang, L, Wen, K-S, Ruan, X, Zhao, Y-X, Wei, F and Wang, Q (2018) Response of plant secondary metabolites to environmental factors. Molecules 23, 762.



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed