IN VITRO EFFECTS OF HYDRO-METHANOLIC EXTRACT FROM Gliricidia sepium LEAVES ON LARVAE OF Haemonchus contortus

Authors

  • Jose Eduardo Garcia Autonomous Agrarian University Antonio Narro, Department of Animal Nutrition, Saltillo 25315, Mexico
  • Leonides Gomez Autonomous Agrarian University Antonio Narro, Department of Animal Nutrition, Calzada Antonio Narro 1923, Saltillo 25315, Mexico
  • Ulises Macias-Cruz Autonomous University of Baja California, Institute of Agricultural Science, Ejido Nuevo León, Mexicali, 21705, Mexico
  • Leonel Avendaño-Reyes Autonomous University of Baja California, Institute of Agricultural Science, Ejido Nuevo León, Mexicali, 21705, Mexico
  • Miguel Mellado * Autonomous Agrarian University Antonio Narro, Department of Animal Nutrition, Calzada Antonio Narro 1923, Saltillo 25315, Mexico, mmellbosq@yahoo.com

DOI:

https://doi.org/10.26873/SVR-1291-2023

Abstract

The aim of this in vitro study was to evaluate the anthelmintic effects of extracts of Gliricidia sepium on sheathed and exsheathed larvae of Haemonchus contortus. Larvae of this parasite were incubated at 20–25 °C in hydro-methanolic extracts of leaves from this tropical tree at concentrations of 12.5, 25, 50, 100, and 200 mg/mL for 24, 48, or 72 h. Water and ivermectin were negative and positive controls, respectively. Total phenolic compounds of leaves of G. sepium were 6.4 ± 2.4 mg/g of dry matter. Other compounds identified in this leguminous tree by HPLC-mass spectrometry and that may be responsible for the anthelmintic effects observed were vanillin 4-sulfate, prodelphinidin p-coumaroyl glucose, kaempferol 3-o-glucosyl-rhamnosyl-glucoside,kaempferol-3-O-xylosyl rutinoside, p-coumaric acid, luteolin 7-rutinoside, isorhamnetin 3-glucoside-7-rhamnoside, and dihydro ferulic acid. At doses of 100 mg/mL mortality rate of sheathed and exsheathed H. contortus was 21.6 and 44.7%, respectively for 72 h of incubation. At 200 mg/mL, the hydro-methanolic extracts of G. sepium killed 61.5 and 93.8% of sheathed and exsheathed larvae, respectively, after 72 h of incubation. The effective concentration of the G. sepium extract for 50% sheathed and exsheathed larvae mortality (EC50) after 72 h of incubation was 74 mg/mL (CI = 46–100) and 68 mg/mL (CI= 32–100), respectively. The significant (P<0.001) ability to kill larvae compared to the negative controls, suggests in vitro anthelmintic properties of G. sepium against H. contortus.

UČINKI HIDROMETANOLNEGA IZVLEČKA LISTOV Gliricidia sepium NA LIČINKE Haemonchus contortus IN VITRO 

Izvleček: Namen te in vitro študije je bil oceniti protiglivične učinke izvlečkov Gliricidia sepium na ličinke Haemonchus contortus z ovojem in brez njega. Ličinke parazita so bile inkubirane 24, 48 ali 72 ur pri 20–25 °C v hidrometanolnih izvlečkih listov tega tropskega drevesa v koncentracijah 12,5, 25, 50, 100 in 200 mg/ml. Voda in ivermectin sta služila kot negativna in pozitivna kontrola. Skupne fenolne spojine v listih G. sepium so obsegale 6,4 ± 2,4 mg/g suhe snovi. Druge spojine, ki so bile v drevesu identificirane s HPLC-masno spektrometrijo in ki bi lahko bile odgovorne za opažene protiglivične učinke, so bile vanilin 4-sulfat, prodelfinidin p-kumaroil glukoza, kaempferol 3-O-glukozil-rimnozil-glukozid, kaempferol-3-O-ksilozil rutinozid, p-kumarna kislina, luteolin 7-rutinozid, izorhamnetin 3-glukozid-7-rimnozid in dihidro ferulinska kislina. Pri odmerkih 100 mg/ml in 72 urah inkubacije je bila stopnja smrtnosti pri H. contortus z ovojem 21,6 %, pri H. contortus brez ovoja pa 44,7 %. Pri odmerkih 200 mg/ml in 72 urah inkubacije so hidrometanolni izvlečki G. sepium uničili 61,5 % ličink z ovojem in 93,8 % ličink brez ovoja. Srednja efektivna koncentracija (EC50) izvlečka G. sepium za ličinke z ovojem je bila 74 mg/ml (CI  =  46–100), za ličinke brez ovoja pa 68 mg/ml (CI  =  32–100) po 72 urah inkubacije. Statistično značilna (P < 0,001) sposobnost uničenja ličink v primerjavi z negativno kontrolo kaže na protiglivične lastnosti G. sepium proti H. contortus in vitro.

Ključne besede: flavonoidi; flavonoli; ličinke; nematodi; tanini

References

● 1. Emery DL, Hunt PW, Le Jambre LF. Haemonchus contortus: the then and now, and where to from here? Int J Parasitol 2016; 46: 755–69.

● 2. Yacob CH, Mistre AH, Adem AH, Basu AK. Parasitological and clinical responses of lambs experimentally infected with Haemonchus contortus (L3) with and without ivermectin treatment.

Vet Parasitol 2009; 166: 119–23.

● 3. Attindehou S, Salifou S, Biaou CF, Gbati OB, Adamou-N´Diaye M, Pangui LJ. Epidemiology of haemonchosis in sheep and goats in Benin. J Parasitol Vector Biol 2012; 4: 20–4.

● 4. Saminathan M, Gopalakrishnan A, Latchumikanthan A, et al. Histopathological and parasitological study of blood-sucking Haemonchus contortus infection in sheep.

Adv Anim Vet Sci 2015; 3: 99–108.

● 5. Fentahun T, Luke G. Small ruminant haemonchosis: prevalence and associated determinants in randomly selected restaurants and hotels of Gondar Town, Ethiopia.

Eur J Appl Sci 2012; 4: 168–72.

● 6. Vineer HR, Steiner J, Knapp-Lawitzke F, et al. Implications of between-isolate variation for climate change impact modelling of Haemonchus contortus populations.

Vet Parasitol 2016; 166: 119–23.

● 7. Zvinorovaa PI, Halimani TE, Muchadeyi FC, Matika O, Riggio V, Dzama K. Prevalence and risk factors of gastrointestinal parasitic infections in goats in low-input low-output farming systems in Zimbabwe. Small Rumin Res 2016; 143: 75–83.

● 8. Rodríguez-Vivas RI, Grisi L, Pérez-De León AA, et al. Potential economic impact assessment for cattle parasites in Mexico. Review. Rev Mex Cienc Pecu 2017; 8: 61–74.

● 9. Geurden T, Chartier C, Fanke J, et al. Anthelmintic resistance to ivermectin and moxidectin in gastrointestinal nematodes of cattle in Europe. Int J Parasitol Drugs Drug Resist 2015; 5: 163–71.

● 10. Baiak BHB, Lehnen CR, da Rocha RA. Anthelmintic resistance in cattle: a systematic review and meta-analysis. Livest Sci 2018; 217: 127–35.

● 11. Kaplan RM. Biology, epidemiology, diagnosis, and management of anthelmintic resistance in gastrointestinal nematodes of livestock. Vet Clin North Am Food Anim Pract 2020; 36: 17–30.

● 12. Muchiut SM, Fernández AS, Steffan PE, Riva E, Fiel CA. Anthelmintic resistance: management of parasite refugia for Haemonchus contortus through the replacement of resistant with susceptible populations. Vet Parasitol 2018; 254: 43–8.

● 13. Sangster NC, Cowling A, Woodgate RG. Ten events that defined anthelmintic resistance research. Trends Parasit 2018; 34: 553–63.

● 14. Basto-Estrella G, Rodríguez-Vivas RI, Delfín-González H, Reyes-Novelo E. Dung beetle (Coleoptera: Scarabaeinae) diversity and seasonality in response to use of macrocyclic lactones at cattle ranches in the Mexican neotropics. Insect Conserv Divers 2014; 7: 73–81.

● 15. Ramos-Trejo OS, Canul-Solís JR, Alvarado-Canché A del R, et al. Growth, forage yield and quality of Morus alba L and Gliricidia sepium Jacq Walp in mixed and pure fodder bank systems in Yucatan, Mexico. Agrofor Syst 2020; 94: 151–7.

● 16. Atapattu AAAJ, Pushpakumara DKNG, Rupasinghe WMD, Senarathne SHS, Raveendra SAST. Potential of Gliricidia sepium as a fuelwood species for sustainable energy generation in Sri Lanka. Agric Res J 2017; 54: 34–9.

● 17. Villanueva-López G, Casanova-Lugo F, Martínez-Zurimendi P, Parsons D, Aguilar-Solís LA. Effect of live fences of Gliricidia sepium on CO2 fluxes in tropical livestock systems. Soil Use Manag 2016; 32: 553–64.

● 18. Kaba JS, Zerbe S, Agnolucci M, et al. Atmospheric nitrogen fixation by gliricidia trees Gliricidia sepium Jacq Kunth ex Walp intercropped with cocoa Theobroma cacao L.

Plant Soil 2019; 435: 323–36.

● 19. González-Villalobos D, Palomares-Naveda R, Navarro E, Razz R, Soto-Castillo G, Quintero Moreno A. The use of Gliricidia sepium in the supplementary feeding of crossbred female calves. Rev Cientif Fac Cien Vet 2002; 12: 384–7.

● 20. Castrejón-Pineda FA, Martínez-Pérez P, Corona, L Cerdán JLV, Mendoza GD. Partial substitution of soybean meal by Gliricidia sepium or Guazuma ulmifolia leaves in the rations of growing lambs. Trop Anim Health Prod 2016; 48: 133–7.

● 21. Romero N, Areche C, Cubides-Cárdenas J, et al. In vitro anthelmintic evaluation of Gliricidia sepium, Leucaena leucocephala, and Pithecellobium dulce: fingerprint analysis of extracts by UHPLC-orbitrap mass spectrometry. Molecules 2020; 25: e3002. doi:103390/molecules25133002

● 22. Wabo Poné J, Kenne Tameli F, Mpoame M, Pamo Tedonkeng E, Bilong Bilong CF. In vitro activities of acetonic extracts from leaves of three forage legumes (Calliandra calotyrsus, Gliricidia sepium and Leucaena diversifolia) on Haemonchus contortus. Asian Pac J Trop Med 2011; 4: 125–8.

● 23. Romero N, Areche C, Cubides-Cárdenas J, Escobar N, García-Beltrán O, Simirgiotis MJ, Céspedes Á. In vitro anthelmintic evaluation of Gliricidia sepium, Leucaena leucocephala, and Pithecellobium dulce: Fingerprint analysis of extracts by uhplc-orbitrap mass spectrometry. Molecules 2020; 25(13): 3002. Ista referenca kot 21

● 24. Mushonga B, Habumugisha D, Kandiwa E, et al. Prevalence of Haemonchus contortus infections in sheep and goats in Nyagatare district, Rwanda. J Vet Med 2018; e3602081.

doi: 10.1155/2018/3602081

● 25. Mpofu TJ, Nephawe KA, Mtileni B. Prevalence and resistance to gastrointestinal parasites in goats: a review. Vet World 2022; 15: 2442–52.

● 26. Stampa E, Schipmann-Schwarze C, Hamm U. Consumer perceptions, preferences, and behavior regarding pasture-raised livestock products: a review. Food Qual Prefer 2020; 82: e103872

doi: 101016/jfoodqual2020103872

● 27. Swain T, Hillis WE. The phenolic constituents of Prunus domestica. I The quantitative analysis of phenolic constituents. J Sci Food Agric 1959; 10: 63–8.

● 28. Taga M, Miller E, Pratt D. Chia seeds as a source of natural lipid antioxidants.

J Am Oil Chem Soc 1984; 61: 928–31.

● 29. Still WC, Kahn M, Mitra A. Rapid chromatographic technique for preparative separations with moderate resolution. J Org Chem 1978; 43: 2923–5.

● 30. Ascacio-Valdés J, Burboa E, Aguilera-Carbo AF, et al. Antifungal ellagitannin isolated from Euphorbia antisyphilitica Zucc. Asian Pac J Trop Biomed 2013; 3: 41–6.

● 31. Muchiut S, Fernández S, Domínguez P, et al. Influence of faecal culture media and incubation time on the yield of infective larvae of Haemonchus contortus (Rudolphi 1803).

Parasitol Res 2021; 120: 1493–7.

● 32. Molina-Botero IC, Arroyave-Jaramillo J, Valencia-Salazar S, et al. Effects of tannins and saponins contained in foliage of Gliricidia sepium and pods of Enterolobium cyclocarpum on fermentation, methane emissions and rumen microbial population in crossbred heifers.

Anim Feed Sci Technol 2019; 251: 1–11.

● 33. Wabo Pone J, Kenne Tameli F, Mbida M, Pamo Tedonkeng E, Bilong Bilong CF. In vitro activities of acetonic extracts from leaves of three forage legumes Calliandra calotyrsus, Gliricidia sepium and Leucaena diversifolia on Haemonchus contortus. Asian Pac J Trop Med 2011; 4: 125–8.

● 34. Von Son-de Fernex E, Alonso-Díaz MA, Valles-de la Mora B, Capetillo-Leal CM. In vitro anthelmintic activity of five tropical legumes on the exsheathment and motility of Haemonchus contortus infective larvae. Exp Parasitol 2012; 131: 413–8.

● 35. Geary TG, Hosking BC, Skuce PJ, et al. World association for the advancement of veterinary parasitology (W.A.A.V.P.) guideline: anthelmintic combination products targeting nematode infections of ruminants and horses. Vet Parasitol 2012; 190: 306–16.

● 36. Hounzangbe-Adote MS, Paolini V, Fouraste I, Moutairou K, Hoste H. In vitro effects of four tropical plants on three lifecycle stages of the parasitic nematode, Haemonchus contortus.

Res Vet Sci 2005; 78: 155–60.

● 37. García JE, Gómez L, Mendoza-de-Gives P, et al. Anthelmintic efficacy of hydro-methanolic extracts of Larrea tridentata against larvae of Haemonchus contortus.

Trop Anim Health Prod 2018; 50: 1099–105.

● 38. Cushnie TPT, Lamb AJ. Antimicrobial activity of flavonoids.

Int J Antimicrob Agents 2005; 26: 343–56.

● 39. Delgado-Núñez EJ, Zamilpa, A, González-Cortazar M, et al. Isorhamnetin: a nematocidal flavonoid from Prosopis laevigata leaves against Haemonchus contortus eggs and larvae. Biomolecules 2020; 10: e773. doi: 101016/jjep2019112402

● 40. Zarza-Albarrán MA, Olmedo-Juárez A, Rojo-Rubio R, et al. Galloyl flavonoids from Acacia farnesiana pods possess potent anthelmintic activity against Haemonchus contortus eggs and infective larvae. J Ethnopharmacol 2020; 249: e112402. doi: 10.1016/j.jep.2019.112402

● 41. Klongsiriwet C, Quijada J, Williams AR, Mueller-Harvey I, Williamson EM, Hoste H. Synergistic inhibition of Haemonchus contortus exsheathment by flavonoid monomers and condensed tannins. Int J Parasitol Drugs Drug Resist 2015; 5: 127–34.

Downloads

Published

2023-07-11

How to Cite

Garcia, J. E., Gomez, L., Macias-Cruz, U., Avendaño-Reyes, L., & Mellado *, M. (2023). IN VITRO EFFECTS OF HYDRO-METHANOLIC EXTRACT FROM Gliricidia sepium LEAVES ON LARVAE OF Haemonchus contortus. Slovenian Veterinary Research, 60(3), 127–34. https://doi.org/10.26873/SVR-1291-2023

Issue

Section

Original Research Article

Most read articles by the same author(s)