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Anthelmintic-Like Activity and Ultrastructure Changes Produced by Two Polyphenolic Combinations against Cooperia punctata Adult Worms and Infective Larvae

Cooperia punctata is one of the most prevalent gastrointestinal nematodes affecting cattle under grazing conditions, and the increasing reports of anthelmintic resistance forces researchers to look for novel control measures. Previous reports have proposed the use of polyphenolic compound (PC) combi...

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Autores principales: von Son-de Fernex, Elke, Zúñiga-Olivos, Estefanía, Jiménez-García, Luis Felipe, Mendoza-de Gives, Pedro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10223250/
https://www.ncbi.nlm.nih.gov/pubmed/37242414
http://dx.doi.org/10.3390/pathogens12050744
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author von Son-de Fernex, Elke
Zúñiga-Olivos, Estefanía
Jiménez-García, Luis Felipe
Mendoza-de Gives, Pedro
author_facet von Son-de Fernex, Elke
Zúñiga-Olivos, Estefanía
Jiménez-García, Luis Felipe
Mendoza-de Gives, Pedro
author_sort von Son-de Fernex, Elke
collection PubMed
description Cooperia punctata is one of the most prevalent gastrointestinal nematodes affecting cattle under grazing conditions, and the increasing reports of anthelmintic resistance forces researchers to look for novel control measures. Previous reports have proposed the use of polyphenolic compound (PC) combinations (Coumarin:Quercetin (CuQ) and Caffeic-acid:Rutin (CaR)) against free-living stages (L(3)) of C. punctata. The objective of this study was to assess the in vitro motility inhibition of C. punctata adult worms and infective larvae using the Larval Motility Inhibition Assay (LMIA) and Adult Motility Inhibition Assay (AMIA), and to assess the structural and ultrastructural changes induced by both treatments using Scanning and Transmission Electron Microscopy. For the LMIA, infective larvae were incubated for 3 h in 0.8 mg mL(−1) and 0.84 mg mL(−1) of CuQ and CaR, respectively. For AMIA, six concentrations and five incubation periods (2, 4, 6, 12 and 24 h) were assessed using each PC combination. Cooperia punctata motility was calculated as a percentage and corrected using control motility percentages. A multiple comparisons Brown–Forsythe and Welch ANOVA test was used to compare larval motility; and to fit the dose–response in AMIA, data were analyzed with a non-linear regression four-parameter logistic equation with a variable slope, using the computer program GraphPad Prism(®) V.9.2.0. Although larval motility was barely affected by both treatments (p > 0.05), adult worm motility was inhibited 100% and 86.9% after 24 h incubation with CuQ and CaR, respectively (p < 0.05). The best fit EC(50) for adult worm motility inhibition were 0.073 ± 0.071 mg mL(−1) and 0.051 ± 0.164 mg mL(−1) for CuQ and CaR, respectively. Main structural and ultrastructural lesions observed in both biological stages were: (i) L(3) sheath–cuticle complex disruption, (ii) collagen fibers degradation; (iii) hypodermic detachment, (iv) seam cell apoptosis and (v) mitochondrial swelling. The alterations observed suggest that the PC combinations interfere with the anatomy and physiology of the locomotive apparatus of the nematodes.
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spelling pubmed-102232502023-05-28 Anthelmintic-Like Activity and Ultrastructure Changes Produced by Two Polyphenolic Combinations against Cooperia punctata Adult Worms and Infective Larvae von Son-de Fernex, Elke Zúñiga-Olivos, Estefanía Jiménez-García, Luis Felipe Mendoza-de Gives, Pedro Pathogens Article Cooperia punctata is one of the most prevalent gastrointestinal nematodes affecting cattle under grazing conditions, and the increasing reports of anthelmintic resistance forces researchers to look for novel control measures. Previous reports have proposed the use of polyphenolic compound (PC) combinations (Coumarin:Quercetin (CuQ) and Caffeic-acid:Rutin (CaR)) against free-living stages (L(3)) of C. punctata. The objective of this study was to assess the in vitro motility inhibition of C. punctata adult worms and infective larvae using the Larval Motility Inhibition Assay (LMIA) and Adult Motility Inhibition Assay (AMIA), and to assess the structural and ultrastructural changes induced by both treatments using Scanning and Transmission Electron Microscopy. For the LMIA, infective larvae were incubated for 3 h in 0.8 mg mL(−1) and 0.84 mg mL(−1) of CuQ and CaR, respectively. For AMIA, six concentrations and five incubation periods (2, 4, 6, 12 and 24 h) were assessed using each PC combination. Cooperia punctata motility was calculated as a percentage and corrected using control motility percentages. A multiple comparisons Brown–Forsythe and Welch ANOVA test was used to compare larval motility; and to fit the dose–response in AMIA, data were analyzed with a non-linear regression four-parameter logistic equation with a variable slope, using the computer program GraphPad Prism(®) V.9.2.0. Although larval motility was barely affected by both treatments (p > 0.05), adult worm motility was inhibited 100% and 86.9% after 24 h incubation with CuQ and CaR, respectively (p < 0.05). The best fit EC(50) for adult worm motility inhibition were 0.073 ± 0.071 mg mL(−1) and 0.051 ± 0.164 mg mL(−1) for CuQ and CaR, respectively. Main structural and ultrastructural lesions observed in both biological stages were: (i) L(3) sheath–cuticle complex disruption, (ii) collagen fibers degradation; (iii) hypodermic detachment, (iv) seam cell apoptosis and (v) mitochondrial swelling. The alterations observed suggest that the PC combinations interfere with the anatomy and physiology of the locomotive apparatus of the nematodes. MDPI 2023-05-22 /pmc/articles/PMC10223250/ /pubmed/37242414 http://dx.doi.org/10.3390/pathogens12050744 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
von Son-de Fernex, Elke
Zúñiga-Olivos, Estefanía
Jiménez-García, Luis Felipe
Mendoza-de Gives, Pedro
Anthelmintic-Like Activity and Ultrastructure Changes Produced by Two Polyphenolic Combinations against Cooperia punctata Adult Worms and Infective Larvae
title Anthelmintic-Like Activity and Ultrastructure Changes Produced by Two Polyphenolic Combinations against Cooperia punctata Adult Worms and Infective Larvae
title_full Anthelmintic-Like Activity and Ultrastructure Changes Produced by Two Polyphenolic Combinations against Cooperia punctata Adult Worms and Infective Larvae
title_fullStr Anthelmintic-Like Activity and Ultrastructure Changes Produced by Two Polyphenolic Combinations against Cooperia punctata Adult Worms and Infective Larvae
title_full_unstemmed Anthelmintic-Like Activity and Ultrastructure Changes Produced by Two Polyphenolic Combinations against Cooperia punctata Adult Worms and Infective Larvae
title_short Anthelmintic-Like Activity and Ultrastructure Changes Produced by Two Polyphenolic Combinations against Cooperia punctata Adult Worms and Infective Larvae
title_sort anthelmintic-like activity and ultrastructure changes produced by two polyphenolic combinations against cooperia punctata adult worms and infective larvae
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10223250/
https://www.ncbi.nlm.nih.gov/pubmed/37242414
http://dx.doi.org/10.3390/pathogens12050744
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