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Lysiloma acapulcensis leaves contain anthelmintic metabolites that reduce the gastrointestinal nematode egg population in sheep faeces

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Abstract

One metabolite against lamb gastrointestinal nematodes (GIN) was identified from extracts of Lysiloma acapulcensis leaves. Firstly, an aqueous extract of L. acapulcensis leaves and two fractions, organic and aqueous, were evaluated at different concentrations against Haemonchus contortus eggs and histotrophic larvae. In other experiment, 35 Pelibuey GIN-infected lambs were randomly divided into five groups, as follows: (1) ivermectin (0.2 mg/kg BW); (2) control (untreated); (3) rutin (10 mg/kg BW, as a flavonoid comparison control); (4) L. acapulcensis leaves ethyl acetate fraction (EtAc-F, 25 mg/kg BW); and (5) L. acapulcensis leaves (dried and grinded, 5 g/kg BW). The EtAc-F fraction caused 94.8% egg-hatching inhibition (P < 0.05, 6.25 mg/ml). Likewise, at 50 mg/ml, this fraction resulted in 100% larval mortality. In the second experiment, the highest eggs per gram (EPG) of faeces reduction caused by ivermectin was 80.2% (ȳ = 61.8%); meanwhile, rutin showed a highest EPG reduction = 66.2% (ȳ = 37.5%). Similarly, EtAc-F fraction showed 62.9% EPG highest reduction (ȳ = 32.5%). On the other hand, lambs fed with L. acapulcensis leaves showed a maximum EPG reduction = 50.1% (ȳ = 29.4%). Chromatography analysis (HPLC; NRM) of EtAc-F fraction revealed the presence of myricitrin as a major compound. Lysiloma acapulcensis leaves EtAc-F exhibited the highest in vitro activity against H. contortus eggs and larvae. The low efficacy of ivermectin suggests the presence of anthelmintic resistance. Lysiloma acapulcensis leaves and fractions can be considered in future experiments searching for a sustainable alternative of GIN control.

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Notes

  1. Note: rutin is a flavonoid with anthelmintic activity and it was included in this study as a control of comparison. The dose of this metabolite was estimated based on a bioavailability study in neonatal calves after oral administration of quercetin, aglycone, or rutin (Maceij et al. 2015).

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Acknowledgments

This research formed part of a retention program of the Dr. Agustín Olmedo-Juárez by CONACYT-México, through the Researcher Retaining Program, 2015.

Funding

This study received financial support from CONACYT-Mexico (Project number 251244), INIFAP (Project number 9454133183, through the Researcher Retaining Program), and Red Temática de Farmoquímicos, CONACYT (Project number 271520).

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Authors and Affiliations

  1. Centro de Investigación Biomédica del Sur, Instituto Mexicano del Seguro Social, Argentina No. 1. Col. Centro. CP, 62790, Xochitepec, Morelos, Mexico

    Manases González-Cortazar & Alejandro Zamilpa

  2. Centro Nacional de Investigación Disciplinaria en Parasitología Veterinaria, INIFAP, Carr. Fed. Cuernavaca-Cuautla, No. 8534, Progreso CP, 62550, Jiutepec, Morelos, Mexico

    María Eugenia López-Arellano, Liliana Aguilar-Marcelino, David Emanuel Reyes-Guerrero, Gabriel Ramírez-Vargas, Agustín Olmedo-Juárez & Pedro Mendoza-de-Gives

  3. Sitio Experimental “Las Margaritas”, INIFAP, Hueytamalco, Puebla, Mexico

    Sara Olazarán-Jenkins

Authors
  1. Manases González-Cortazar
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  2. Alejandro Zamilpa
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  3. María Eugenia López-Arellano
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  4. Liliana Aguilar-Marcelino
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  5. David Emanuel Reyes-Guerrero
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  6. Sara Olazarán-Jenkins
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  7. Gabriel Ramírez-Vargas
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  8. Agustín Olmedo-Juárez
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  9. Pedro Mendoza-de-Gives
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Corresponding authors

Correspondence to Agustín Olmedo-Juárez or Pedro Mendoza-de-Gives.

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Ethical approval

All animal experiments strictly complied with the guidelines under the Norma Official Mexicana (Official Rule Number) NOM-051-ZOO-1995 (http://www.senasica.gob.mx) as well as the LEY FEDERAL DE SANIDAD ANIMAL (Federal law for animal health) DOF 07-06-2012 (http://diputados.gob.mx/LeyesBibliop/ref./lfsa.htm), which dictates that all procedures performed in studies involving animals be accomplished by strictly following the Federal Law and Official Rule cited above in accordance with the ethical standards at INIFAP and based in part upon the “Guide for the Care and Use of Laboratory Animals” published by the Institute of Laboratory Animals Resources Commission on Life Sciences, National Research Council, 1996.

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The authors declare that they have no conflict of interest.

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González-Cortazar, M., Zamilpa, A., López-Arellano, M.E. et al. Lysiloma acapulcensis leaves contain anthelmintic metabolites that reduce the gastrointestinal nematode egg population in sheep faeces. Comp Clin Pathol 27, 189–197 (2018). https://doi.org/10.1007/s00580-017-2577-1

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