Cheryl & ethnovet

Monday, 2 January 2017

Établissement du profil chromatographique liquide non aqueux des métabolites phytochimiques apolaires des phytomédicaments

Comptes Rendus Chimie

Volume 19, Issue 7, July 2016, Pages 863–875

Chemistry and Natural Resources/Chimie et ressources naturelles

Mémoire/Full paper

Establishing the non-aqueous liquid chromatographic profile of non-polar metabolites of herbal medicinal products

^a Département de chimie analytique, chimie minérale et générale, unité de formation et de recherche (UFR) des sciences pharmaceutiques et biologiques, université félix houphouet boigny d‘Abidjan, Cocody 01 BP V 34 Abidjan, Côte d’Ivoire
^b Groupe de chimie analytique de Paris-Sud, EA4041 IFR141, faculté de pharmacie, université Paris-Sud, 5, rue Jean-Baptiste-Clément, 92296 Châtenay-Malabry cedex, France
^c Laboratoire de pharmacognosie, CNRS UMR 8076 BioCIS, LabEX LERMIT, faculté de pharmacie, Univ. Paris-Sud, 5, rue Jean-Baptiste-Clément, 92296 Châtenay-Malabry cedex, France

Received 3 June 2015, Accepted 11 February 2016, Available online 11 April 2016

http://dx.doi.org/10.1016/j.crci.2016.02.010

Résumé

Le profil chromatographique des métabolites phytochimiques constitue un outil pour le contrôle de qualité des phytomédicaments africains. Notre objectif était de proposer un protocole de préparation d’échantillon et d’établissement de profil chromatographique liquide des métabolites apolaires pour l’évaluation de la qualité de phytomédicaments africains. La méthodologie était basée sur le traitement chimiométrique des profils chromatographiques des métabolites apolaires issus de différents lots de feuilles de Combretum micranthum et de Mitracarpus scaber. Le profilage métabolique est effectué par chromatographie liquide sur carbone graphite poreux en milieu non aqueux, couplée à la spectrométrie de masse, après extraction au dichlorométhane et élimination de la chlorophylle. Notre méthode d'analyse par chromatographie liquide couplée à la spectrométrie de masse permet de détecter les métabolites apolaires déjà identifiés dans les deux drogues végétales. L'analyse chimiométrique des données des profils chromatographiques utilisant l’analyse PLS-discriminante, avec ou sans correction orthogonale du signal, permet une distinction entre les deux drogues végétales.

Abstract

Chromatographic profiling of plant metabolites is therefore a good tool for quality control of such herbal medicinal products. Our objective was to propose a protocol for sample preparation and liquid chromatographic profiling of non-polar metabolites for quality assessment of African herbal medicinal products. The methodology is based on the chemometric assessment of liquid chromatographic profiles of non-polar metabolites issued from several batches of leaves of Combretum micranthum and Mitracarpus scaber. Metabolic profiling is carried out by non-aqueous liquid chromatography on porous carbon graphite, coupled with mass spectrometry, after extraction with dichloromethane and removal of chlorophyll. Our method using liquid chromatography, coupled to mass spectrometry can detect non-polar metabolites already identified in the two herbal drugs. Chemometric data analysis of chromatographic profiles using the PLS-discriminant analysis with or without orthogonal signal correction, allowed a distinction between the two herbal drugs.

Mots-clés

Profil chromatographique liquide non aqueux;
Spectrométrie de masse;
Chimiométrie;
Métabolites apolaires;
Phytomédicaments;
Combretum micranthum;
Mitracarpus scaber

Keywords

Non-aqueous liquid chromatographic profile;
Mass spectrometry;
Chemometrics;
Non-polar metabolites;
Herbal medicinal products;
Combretum micranthum;
Mitracarpus scaber

: Corresponding author. Département de chimie analytique, chimie minérale et générale, technologie alimentaire, unité de formation et de recherche (UFR) des sciences pharmaceutiques et biologiques, université d‘Abidjan, Cocody 01 BP V 34 Abidjan, Côte d’Ivoire.

Is there nothing new under the sun? The influence of herbals and pharmacopoeias on ethnobotanical traditions in Albacete (Spain)

2017 Jan 4;195:96-117. doi: 10.1016/j.jep.2016.11.040. Epub 2016 Nov 26.

Rivera D¹, Verde A², Obón C³, Alcaraz F⁴, Moreno C⁵, Egea T⁶, Fajardo J⁷, Palazón JA⁸, Valdés A⁹, Signorini MA¹⁰, Bruschi P¹¹.

Author information

¹Depto. Biología Vegetal, Campus de Espinardo, Universidad de Murcia, Murcia, Spain. Electronic address: drivera@um.es.
²Grupo de Investigación en Etnobiología, Flora y Vegetación del Sureste Ibérico. Laboratorio de Sistemática y Etnobotánica, Instituto Botánico, UCLM, Jardín Botánico de Castilla-La Mancha, Avenida de la Mancha s/n, 02006 Albacete, Spain. Electronic address: alonsoverde@gmail.com.
³Dpto. Biología Aplicada, Escuela Politécnica Superior, Universidad Miguel Hernández, Ctra. Beniel Km 3.2, 03312 Orihuela, Alicante, Spain. Electronic address: cobon@umh.es.
⁴Depto. Biología Vegetal, Campus de Espinardo, Universidad de Murcia, Murcia, Spain. Electronic address: falcaraz@um.es.
⁵Archivo Histórico Provincial, Padre Romano, 2, Albacete, Spain. Electronic address: candelariam@jccm.es.
⁶Dpto. Biología Aplicada, Escuela Politécnica Superior, Universidad Miguel Hernández, Ctra. Beniel Km 3.2, 03312 Orihuela, Alicante, Spain; Dipartimento di Scienze delle Produzioni Agroalimentari e dell'Ambiente, Università degli Studi di Firenze, Florence, Italy. Electronic address: t.egeamolines@gmail.com.
⁷Grupo de Investigación en Etnobiología, Flora y Vegetación del Sureste Ibérico. Laboratorio de Sistemática y Etnobotánica, Instituto Botánico, UCLM, Jardín Botánico de Castilla-La Mancha, Avenida de la Mancha s/n, 02006 Albacete, Spain. Electronic address: josefajard@gmail.com.
⁸Dpto. Ecología, Campus de Espinardo, Universidad de Murcia, Murcia, Spain. Electronic address: palazon@um.es.
⁹Grupo de Investigación en Etnobiología, Flora y Vegetación del Sureste Ibérico. Laboratorio de Sistemática y Etnobotánica, Instituto Botánico, UCLM, Jardín Botánico de Castilla-La Mancha, Avenida de la Mancha s/n, 02006 Albacete, Spain. Electronic address: arturo.valdes@uclm.es.
¹⁰Dipartimento di Biologia, Università degli Studi di Firenze, Florence, Italy. Electronic address: msignorini@unifi.it.
¹¹Dipartimento di Scienze delle Produzioni Agroalimentari e dell'Ambiente, Università degli Studi di Firenze, Florence, Italy. Electronic address: piero.bruschi@unifi.it.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE:

This paper has two overarching aims: (1) presenting the results of studying the Albacete tariff of medicines of 1526 and (2) broadly analyzing the origin and influences of medicinal traditional knowledge in the region of Albacete, Spain. We use historical and modern literature that may have influenced this knowledge. Our primary goal was to determine the ingredients used in the pharmacy in the 16th century CE in Albacete through the analysis of the tariff, and our secondary goal was to investigate until when ingredients and uses present in pharmacy and herbals persisted in later periods.

METHODS:

The identity of medicines and ingredients was determined by analyzing contemporary pharmacopoeias and classical pharmaceutical references. We analyzed further 21 sources (manuscripts, herbals, and books of medicines, pharmacopoeias, pharmacy inventories, and modern ethnobotanical records) for the presence/absence of ingredients and complex formulations of the tariff. Using factorial and cluster analysis and Bayesian inference applied to evolution models (reversible-jump Markov chain Monte Carlo), we compared textual sources. Finally, we analyzed the medicinal uses of the top 10 species in terms of frequency of citation to assess the dependence of modern ethnobotanical records on Renaissance pharmacy and herbals, and, ultimately, on Dioscorides.

RESULTS:

In Albacete 1526, we determined 101 medicines (29 simple drugs and 72 compound medicines) comprising 187 ingredients (85% botanical, 7.5% mineral, and 7.5% zoological substances). All composed medicines appear standardized in the pharmacopoeias, notably in the pharmacopoeia of Florence from 1498. However, most were no longer in use by 1750 in the pharmacy, and were completely absent in popular herbal medicine in Albacete 1995 as well as in Alta Valle del Reno (Italy) in 2014. Among the ingredients present in different formulation are the flowers of Rosa gallica, honey (Apis mellifera), the roots of Nardostachys jatamansi, and Convolvulus scammonia, pistils of Crocus sativus, grapes and raisins (Vitis vinifera), rhizomes of Zingiber officinale, bark of Cinnamomum verum, leaves and fruits of Olea europaea, mastic generally of Pistacia lentiscus, and wood of Santalum album. The statistical analysis of sources produces four well-separated clusters (Renaissance Herbals and Pharmacopoeias, Ethnobotany and Folk Medicine, Old phytotherapy, and Modern phytotherapy including Naturopathy) confirming our a priori classification. The clade of Renaissance Herbals and Pharmacopoeias appears separated from the rest in 97% of bootstrapped trees. Bayesian inference produces a tree determined by an initial set of two well-distinct core groups of ingredients: 64, locally used in Mediterranean Europe during centuries; and 45, imported, used in pharmacy during centuries. Complexity reached its maximum in Albacete 1526 and contemporary pharmacopoeias, gradually decreasing over time. The analysis of medicinal uses of the top 10 ingredients showed low coincidence between Dioscorides and different Renaissance herbals or medical treatises and of all of them with ethnobotany in Albacete.

CONCLUSIONS:

Regarding our question: is there something new under the sun? In some aspects, the answer is "No". The contrast between expensive drugs, highly valued medicines, and unappreciated local wild medicinal plants persists since the Salerno's school of medicine. Old medicine in Mediterranean Europe, as reflected by Albacete 1526 tariff of medicines, involved strict formulations and preferences for certain ingredients despite other ingredients locally available but underappreciated. This confirms the fact that any system of medicine does not get to use all available resources. Ethnobiological records of materia medica, in rural areas of Albacete, describe systems with a high degree of stability and resilience, where the use of local resources, largely wild but also cultivated, is predominant in contrast with the weight of imported exotic products in pharmacy.

KEYWORDS:

Ethnobotany; Herbals; Medicinal plants; Multivariate and Bayesian analysis; Pharmacopoeias; Traditional medicine

PMID:: 27894973
DOI:: 10.1016/j.jep.2016.11.040

Regulation of miRNAs by herbal medicine: An emerging field in cancer therapies

2016 Dec 19;86:262-270. doi: 10.1016/j.biopha.2016.12.023. [Epub ahead of print]

Mohammadi A¹, Mansoori B¹, Baradaran B².

Author information

¹Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
²Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: Behzad_im@yahoo.com.

Abstract

MicroRNAs' expression profiles have recently gained major attention as far as cancer research is concerned. MicroRNAs are able to inhibit target gene expression via binding to the 3' UTR of target mRNA, resulting in target mRNA cleavage or translation inhibition. MicroRNAs play significant parts in a myriad of biological processes; studies have proven, on the other hand, that aberrant microRNA expression is, more often than not, associated with the growth and progression of cancers. MicroRNAs could act as oncogenes (oncomir) or tumor suppressors and can also be utilized as biomarkers for diagnosis, prognosis, and cancer therapy. Recent studies have shown that such herbal extracts as Shikonin, Sinomenium acutum, curcumin, Olea europaea, ginseng, and Coptidis Rhizoma could alter microRNA expression profiles through inhibiting cancer cell development, activating the apoptosis pathway, or increasing the efficacy of conventional cancer therapeutics. Such findings patently suggest that the novel specific targeting of microRNAs by herbal extracts could complete the restriction of tumors by killing the cancerous cells so as to recover survival results in patients diagnosed with malignancies. In this review, we summarized the current research about microRNA biogenesis, microRNAs in cancer, herbal compounds with anti-cancer effects and novel strategies for employing herbal extracts in order to target microRNAs for a better treatment of patients diagnosed with cancer.

KEYWORDS:

Coptidis rhizoma; Curcumin; Ginseng; Herbal medicine; Micro RNA; Olea europaea; Shikonin; Sinomenium acutum

PMID:: 28006752
DOI:: 10.1016/j.biopha.2016.12.023

In vitro anthelmintic activity of Zanthoxylum simulans essential oil against Haemonchus contortus.

2015 Jul 30;211(3-4):223-7. doi: 10.1016/j.vetpar.2015.05.029. Epub 2015 Jun 5.

Qi H¹, Wang WX¹, Dai JL¹, Zhu L².

Author information

¹College of Food and Bioengineering, South China University of Technology, Guangzhou, Guangdong Province, China.
²College of Food and Bioengineering, South China University of Technology, Guangzhou, Guangdong Province, China. Electronic address: zhuliang@scut.edu.cn.

Abstract

The need for new anthelmintic agents with low impact on the environment is becoming urgent. Phytotherapy is an alternative method to control gastrointestinal nematodes in small ruminants. This study aims to determine the composition of Zanthoxylum simulans essential oil (ZSEO) and evaluate the in vitro ovicidal and larvicidal effects of ZSEO on Haemonchus contortus using egg hatch assay, larval development assay (LDA), and larval migration inhibition assay (LMIA). The chemical composition of ZSEO was determined through gas chromatography and mass spectrometry and 94 compounds were identified from the ZSEO. The major constituents of ZSEO were borneol (18.61%), β-elemene (10.87%). ZSEO and borneol both at 40 mg/mL inhibited larval hatching by 100%, with LC50 values of 3.98 and 1.50mg/mL, respectively. The LC50 value of β-elemene was not determined because of its insufficient activity. The results of LDA showed that ZSEO, borneol, and β-elemene all at 40 mg/mL inhibited larval development by 99.8%, 100%, and 55.4%, respectively, and exhibited dose-dependent responses with LC50 values of 4.02, 1.99, and 32.17 mg/mL, respectively. The results of LMIA showed that ZSEO, borneol, and β-elemene all at 40 mg/mL inhibited larval migration by 74.3%, 97.0%, and 53.2%, respectively. ZSEO presented ovicidal and larvicidal activities in vitro. Therefore, Zanthoxylum may be an alternative source of anthelmintic agents to control gastrointestinal nematodes in sheep.

KEYWORDS:

Anthelmintic activity; Composition; Essential oil; Zanthoxylum simulans

PMID:: 26073109
DOI:: 10.1016/j.vetpar.2015.05.029

[PubMed - indexed for MEDLINE]

In vivo activity of albendazole in combination with thymol against Echinococcus multilocularis.

2015 Sep 15;212(3-4):193-9. doi: 10.1016/j.vetpar.2015.06.030. Epub 2015 Jul 10.

Albani CM¹, Pensel PE¹, Elissondo N², Gambino G², Elissondo MC³.

Author information

¹Laboratorio de Zoonosis Parasitarias, Departamento de Biología, Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Mar del Plata (UNMdP), Funes 3350, 7600 Mar del Plata, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
²Laboratorio de Análisis Clínicos Santisteban, 7000 Tandil, Buenos Aires, Argentina.
³Laboratorio de Zoonosis Parasitarias, Departamento de Biología, Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Mar del Plata (UNMdP), Funes 3350, 7600 Mar del Plata, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina. Electronic address: mceliss@mdp.edu.ar.

Abstract

Human alveolar echinococcosis (AE) is caused by the fox tapeworm Echinococcus multilocularis and is usually lethal if left untreated. The current strategy for treating human AE is surgical resection of the parasite mass complemented by chemotherapy with benzimidazole compounds. However, reliable chemotherapeutic alternatives have not yet been developed stimulating the research of new treatment strategies such as the use of medicinal plants. The aim of the current study was to investigate the efficacy of the combination albendazole (ABZ)+thymol on mice infected with E. multilocularis metacestodes. For this purpose, mice infected with parasite material were treated daily for 20 days with ABZ (5 mg/kg), thymol (40 mg/kg) or ABZ (5 mg/kg)+thymol (40 mg/kg) or left untreated as controls. After mice were euthanized, cysts were removed from the peritoneal cavity and the treatment efficacy was evaluated by the mean cysts weight, viability of protoscoleces and ultrastructural changes of cysts and protoscoleces. The application of thymol or the combination of ABZ+thymol resulted in a significant reduction of the cysts weight compared to untreated mice. We also found that although ABZ and thymol had a scolicidal effect, the combination of the two compounds had a considerably stronger effect showing a reduction in the protoscoleces viability of 62%. These results were also corroborated by optical microscopy, SEM and TEM. Protoscoleces recovered from ABZ or thymol treated mice showed alterations as contraction of the soma region, rostellar disorganization and presence of blebs in the tegument. However both drugs when combined lead to a total loss of the typical morphology of protoscoleces. All cysts removed from control mice appeared intact and no change in ultrastructure was detected. In contrast, cysts developed in mice treated with ABZ revealed changes in the germinal layer as reduction in cell number, while the treatment with thymol or the ABZ+thymol combination predominantly showed presence of cell debris. On the other hand, no differences were found in alkaline phosphatase (AP), glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) activities between control and treated mice, indicating the lack of toxicity of the different drug treatments during the experiment. Because combined ABZ+thymol treatment exhibited higher treatment efficiency compared with the drugs applied separately against murine experimental alveolar echinococcosis, we propose it would be a useful option for the treatment of human AE.

KEYWORDS:

Albendazole; Alveolar echinococcosis; Echinococcus multilocularis; Liver enzymes; Thymol

Efficacy of osthole for Echinococcus granulosus in vitro and Echinococcus multilocularis in vivo

2016 Aug 15;226:38-43. doi: 10.1016/j.vetpar.2016.05.016. Epub 2016 Jun 23.

Yuan M¹, Luo Y², Xin Q³, Gao H³, Zhang G², Jing T⁴.

Author information

¹Institute of Pathogenic Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, China; The Research Institute of Biomedical Nanotechnology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, China.
²Institute of Immunology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, China.
³Institute of Pathogenic Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, China.
⁴Institute of Pathogenic Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, China; The Research Institute of Biomedical Nanotechnology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, China. Electronic address: landa021001@aliyun.com.

Abstract

Echinococcosis is a zoonotic infection caused by cestode species of the genus Echinococcus; in addition, this zoonosis has long been neglected as a parasitic disease and has limited treatment options. Clinical drugs such as benzimidazole derivatives have limited treatment efficacy. The current study evaluated a novel drug, osthole, with low toxicity and high activity against Echinococcus in vitro and in vivo. The results in vitro indicated that the viability of Echinococcus granulosus protoscoleces in the group treated with osthole (120μM) decreased by 100% within 3days. In vivo experiments were conducted using parasite-infected mice. For this purpose, three groups of infected mice were treated daily for 6 weeks with albendazole (ABZ, 100mg/kg, positive control group), osthole (100mg/kg, experimental group), or honey/PBS (100mg/kg, negative control group), respectively. The osthole- and ABZ-treated groups presented a significant reduction in wet weight of metacestodes, increase in the level of interleukin (IL)-4 and the percentage of eosinophils compared with the control group. Osthole exhibited a high activity against echinococcosis in vivo. In addition, the toxicity of osthole was evaluated via an in vitro 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay, as well as via morphological observation and calculation of liver and kidney function indexes in vivo. No obvious toxic effects of osthole were observed in our study. Therefore, this novel drug may be a promising alternative to benzimidazole in anti-echinococcosis chemotherapy.

KEYWORDS:

Echinococcosis; Efficacy; Osthole; Toxicity

Synergistic action of fatty acids, sulphides and stilbene against acaricide-resistant Rhipicephalus microplus ticks

2016 Sep 15;228:121-125. doi: 10.1016/j.vetpar.2016.08.023. Epub 2016 Aug 31.

Arceo-Medina GN¹, Rosado-Aguilar JA², Rodríguez-Vivas RI¹, Borges-Argaez R³.

Author information

¹Laboratorio de Parasitología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma de Yucatán, Carretera Mérida-Xmatkuil Km. 15.5, 97100 Mérida, Yucatán, Mexico.
²Laboratorio de Parasitología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma de Yucatán, Carretera Mérida-Xmatkuil Km. 15.5, 97100 Mérida, Yucatán, Mexico. Electronic address: alberto.rosadoaguilar@gmail.com.
³Centro de Investigación Científica de Yucatán, Calle 43 No. 130, Col. Chuburná de Hidalgo, 97200 Mérida, Yucatán, Mexico.

Abstract

Six compounds in a methanolic extract of Petiveria alliacea stem (cis-stilbene; benzyl disulphide; benzyl trisulphide; and methyl esters of hexadecanoic acid, octadecadienoic acid and octadecenoic acid) are known to exercise acaricide activity against cattle tick Rhipicephalus microplus larvae and adults. The synergistic effect of 57 combinations of these six compounds on acaricide activity against R. microplus was evaluated. Larvae immersion tests produced the lethal concentrations needed to kill 50% (LC₅₀) and 99% (LC₉₉) of the population. Adult immersion tests produced rates (%) for mortality, oviposition inhibition and eclosion inhibition. Individually, none of the compounds (1% concentration) exhibited acaricide activity (mortality ≤2.3%). When combined, however, nine mixtures exhibited a synergistic increase in activity, with high mortality rates (≥92%) in larvae. Values for LC₅₀ ranged from 0.07 to 0.51% and those for LC₉₉ from 0.66 to 5.16%. Thirty six compound mixtures had no significant activity (mortality ≤30%) against larvae. Two mixtures exhibited synergism against adults, with high rates (≥92%) of oviposition inhibition. The mixtures based on the benzyl disulphide+benzyl trisulphide pairing produced a synergistic effect against acaricide-resistant R. microplus larva and adults, and are therefore the most promising combination for controlling this ubiquitous ectoparasite.

KEYWORDS:

Benzyl disulphide; Benzyl trisulphide; Rhipicephalus microplus; Synergism

PMID:: 27692312
DOI:: 10.1016/j.vetpar.2016.08.023

[PubMed - in process]

Acaricidal efficacies of Lippia gracilis essential oil and its phytochemicals against organophosphate-resistant and susceptible strains of Rhipicephalus (Boophilus) microplus.

2016 Sep 15;228:60-64. doi: 10.1016/j.vetpar.2016.05.028. Epub 2016 May 24.

Costa-Júnior LM¹, Miller RJ², Alves PB³, Blank AF⁴, Li AY⁵, Pérez de León AA⁶.

Author information

¹Pathology Department, Federal University of Maranhão, Brazil. Electronic address: livio.martins@ufma.br.
²Cattle Fever Tick Research Laboratory, Agricultural Research Service, USDA, United States.
³Chemistry Department, Federal University of Sergipe, Brazil.
⁴Agronomy Department, Federal University of Sergipe, Brazil.
⁵Invasive Insect Biocontrol and Behavior Laboratory, Agricultural Research Service, USDA, United States.
⁶Knipling-Bushland U.S. Livestock Insects Research Laboratory, and Veterinary Pest Genomics Center, Agricultural Research Service, USDA, United States.

Abstract

Plant-derived natural products can serve as an alternative to synthetic compounds for control of ticks of veterinary and medical importance. Lippia gracilis is an aromatic plant that produces essential oil with high content of carvacrol and thymol monoterpenes. These monoterpenes have high acaricidal activity against Rhipicephalus (Boophilus) microplus. However, there are no studies that show efficacy differences of essential oils between susceptible and organophosphate resistant strains of R. (B.) microplus. The aim of the present study was to compare acaricidal effects of essential oils extracted from two different genotypes of L. gracilis and the main monoterpenes on larvae of both susceptible and organophosphate resistant R. (B.) microplus larvae. The efficacy of the essential oil of two genotypes of L. gracilis (106 and 201) and their monoterpenes carvacrol and thymol was measured using the larval immersion test on coumaphos-resistant and susceptible strains of R. (B.) microplus. Lethal concentrations were calculated using GraphPad Prism 6.0. Chemical analysis was performed by GC-MS and FID. Thymol and carvacrol were observed to be major constituents in 106 and 201L. gracilis genotype essential oils, respectively. Essential oils of both genotypes were more effective against organophosphate-resistant tick strain than susceptible tick strain. Carvacrol was 3.2 times more toxic to organophosphate resistant strain than to susceptible strain. Thymol was equally toxic to resistant and susceptible tick strains. The significantly higher efficacy monoterpene carvacrol against resistant ticks may lead to development of new natural product acaricide formulations for use to control organophosphate resistant R. (B.) microplus populations.

KEYWORDS:

Acaricide; Botanical; Carvacrol; Organophosphate; Resistance; Thymol

Sunday, 1 January 2017

Chemical composition and in vitro activity of Calotropis procera (Ait.) latex on Haemonchus contortus

Veterinary Parasitology

Volume 226, 15 August 2016, Pages 22–25

Short communication

^a Programa de Pós-Graduação em Ciências Veterinárias, Universidade Estadual do Ceará, Brazil
^b Laboratório de Química de Produtos Naturais, Universidade Estadual do Ceará, Brazil
^c Laboratório de Microscopia e Microanálise, Centro de Tecnologias Estratégicas do Nordeste, Brazil

Received 18 November 2015, Revised 1 June 2016, Accepted 7 June 2016, Available online 8 June 2016

http://dx.doi.org/10.1016/j.vetpar.2016.06.012

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Highlights

•: A new compound, 1-(2′,5′-dimethoxyphenyl)-glycerol, was isolated in ethyl acetate extract of Calotropis procera latex (EAECPL).
•: EAECPL efficiently inhibited egg hatching, larval development and motility of the adult worms of Haemonchus contortus.
•: Structural changes were not detected in the worm's cuticle treated with EAECPL.

Abstract

Calotropis procera is among the species of medicinal plants that have traditionally been used for the treatment of parasites in small ruminants, stimulating the scientific validation of anthelmintic effects. This study aimed to investigate the chemical composition of ethyl acetate extract of Calotropis procera latex (EAECPL), assess the in vitro effect against Haemonchus contortus and the structural changes caused in the adult worm. The latex was collected, lyophilized and subjected to washing with the ethyl acetate solvent to obtain EAECPL. The constituents of the extract were isolated by column chromatography and identified by ¹³C and ¹H nuclear magnetic resonance spectroscopy. The egg hatching test (EHT), larval development test (LDT) and adult worms motility test (WMT) were conducted to evaluate the effectiveness of EAECPL on eggs, larvae and adult of H. contortus, respectively. The worms obtained from the WMT, after 24 h exposure to EAECPL or controls were observed on a scanning electron microscope (SEM). The results were analysed by variance analysis and compared with Tukey’s test (P < 0.05). Three compounds were isolated from EAECPL and identified as urs-19(29)-en-3-yl acetate, (3β)-Urs-19(29)-en-3-ol, and 1-(2′,5′-dimethoxyphenyl)-glycerol. In the EHT, EAECPL inhibited larval hatching by 91.8% at dose of 4 mg/ml. In the LDT 1 mg/ml inhibited 99.8% larval development. In the WMT, EAECPL in the concentration of 100 μg/ml inhibited 100% motility of worms, 12 h post-exposition. In the SEM, obvious differences were not detected between the negative control worms and the worms treated with EAECPL. In this study, EAECPL showed an effect on inhibition egg hatching, larval development and motility of the adult worms of H. contortus. This should be related both to the identified compounds, as well as the other compounds present in the EAECPL, acting alone or synergistically.

Keywords

Calotropis procera;
Gastrointestinal nematodes;
Phytotherapy;
Sheep

: Corresponding author at: Programa de Pós-graduação em Ciências Veterinárias/FAVET/UECE, Av. Dr. Silas Munguba, 1700, Campus do Itaperi, CEP 60714−903, Fortaleza, Ceará, Brazil.

Chemical composition and in vitro activity of Calotropis procera (Ait.) latex on Haemonchus contortus

Anti-helminthic activity of Momordica charantia L. against Fasciola hepatica eggs after twelve days of incubation in vitro

Veterinary Parasitology

Volume 228, 15 September 2016, Pages 160–166

Research paper

^a Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas, Gerais, Brazil
^b Departamento de Produtos Farmacêuticos, Faculdade de Fármacia, Universidade Federal de Minas Gerais, Brazil
^c Departamento de Ciências Agrárias, Universidade Estadual de Montes Claros, Brazil
^d Department of Statistics, Universidade Federal de Ouro Preto, Ouro Preto, MG, Brazil

Received 23 May 2016, Revised 19 August 2016, Accepted 31 August 2016, Available online 11 September 2016

http://dx.doi.org/10.1016/j.vetpar.2016.08.025

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Highlights

•: Momordica charantia prevent embryogenesis in Fasciola hepatica eggs.
•: Anti-ovicidal activity against F. hepatica of the flavonoid quercetin M. charantia.
•: Momordica charantia is a potential alternative to control helminth parasites.

Abstract

Fasciolosis, a parasitic disease caused by the trematode Fasciola hepatica underreported is expanding both in human and animal population, throughout the world. The constant use of synthetic drugs to treat this condition has led to the natural selection of resistant strains of the parasite. Hence, there is a growing focus on the potential anti-helminthic properties of medicinal plants and phytopharmaceuticals. The current study assessed the potential anti-fasciolicide action of Momordica charantia leaf extracts and fractions on the eggs of F. hepatica parasites. The lyophilized crude extract (CE) of M. charantia leaves and its sub-fractions, obtained from liquid–liquid partitioning with organic solvents, were analysed by High Performance Liquid Chromatography (HPLC), suspended in 1% DMSO and used in in vitro tests. Quadruplicates of 50 F. hepatica eggs were incubated at 23 °C with M. charantia leaf CE in different concentrations. After 12 days no larvae were formed in eggs incubated with CE concentrations above 12.5 mg/mL. Eggs incubated with CE sub-fractions at concentrations of 1000, 100, 10, 1, 0.1, 0.01 μg/mL affected embryonic development, with n-butanol presenting the strongest inhibition of miracidia formation. In contrast, on the 12th day, 90% of the miracidia hatched in the control experiments using 0.03% DMSO whereas embryogenesis was completely abolished with any concentration of albendazole sulphoxide ABZ(SO). Chemical analysis of the CE and sub-fractions revealed a prominent presence of flavonoids. HPLC–MS confirmed Quercetin to be one of the main flavonoids present in the CE and the n-butanol subfraction. This is the first study to analyse the potential anti-fasciolicide action of M. charantia leaf CE and subfractions.

Graphical abstract

Keywords

Fasciola hepatica;
Momordica charantia L.;
Flavonoid;
Quercetin;
Anti-ovicidal activity

: Corresponding author at: Av. Presidente Antôni°Carlos 6627, zip code 31270−901 Belo Horizonte − MG, Brazil.

Anticoccidial efficacy of naringenin and a grapefruit peel extract in growing lambs naturally-infected with Eimeria spp.

Veterinary Parasitology

Volume 232, 15 December 2016, Pages 58–65

^a Departamento de Parasitología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Mexico City, Mexico
^b Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma del Estado de México, Toluca, Mexico
^c Departamento de Medicina y Zootecnia de Rumiantes, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Mexico City, Mexico

Received 10 July 2016, Revised 7 October 2016, Accepted 8 November 2016, Available online 9 November 2016

Research paper

http://dx.doi.org/10.1016/j.vetpar.2016.11.009

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Highlights

•: Citrus extracts exert an anticoccidial effect.
•: Citrus extracts and toltrazuril have antioxidant properties.
•: Nitric oxide production lowers in lambs after Citrus extracts ingestion.
•: Toltrazuril decreases nitric oxide generation.
•: Anticoccidial efficacy may be related to pharmacokinetic profiles of extracts.

Abstract

The current study aimed to determine the anti-Eimeria efficacy of an extract of grapefruit peels (GF) and commercial naringenin (NAR) in naturally-infected lambs, as well as the influence of these flavonoids on the oxidative status during ovine coccidiosis. Pharmacokinetic profiles were also determined. Extracts were administered per os to Eimeria naturally infected growing lambs during 90 consecutive days. The commercial anticoccidial drug toltrazuril (TTZ) was included in this trial as a standard. Twenty-four lambs were divided into four groups: NAR, lambs given a daily dose of 5 mg of a commercial naringenin extract of 98% higher purity per kg body weight; GF, lambs that recived a daily dose of 5 mg of ethanolic extract of grapefruit peels per kg body weight; TTZ, lambs treated with 20 mg of toltrazuril/kg body weight on days 0 and 15 of the experiment; and CTRL, untreated lambs that received daily dose of 30 ml of water. Daily doses of GF and NAR were dissolved in 30 ml of water and orally given to animals; whereas toltrazuril was administered as a single dose of an undiluted suspension to lambs of the TTZ group. The CTRL group received 30 ml of water; as well as the TTZ group for the period after the single dose administration. Fecal and serum samples were collected from all lambs. Anticoccidial efficacy was estimated by coprological techniques. Generation of nitric oxide levels and the antioxidant capacity of the experimental compounds were determined by the Griess and ABTS assays, respectively. The pharmacokinetic parameters of NAR and the GF extract were obtained. On day 30 post-ingestion, anticoccidial efficacy was 91.76% (NAR) and 89.65% (GF); whereas 99.63% of efficacy was achieved with TTZ 15 days after treatment. NAR, GF and TTZ significantly reduced oxidative stress in infected animals. The mean daily weight gain for each group was 122 g (NAR), 122 g (GF), 143 g (TTZ) and 98 g (CTRL). Following the oral administration of NAR and GF, values in plasma approached maximum concentrations within 2.1 to 2.5 h. In conclusion, the administration of NAR and the GF extract reduced Eimeria oocyst output, oxidative stress and promoted higher mean daily weight gains in infected lambs.

Graphical abstract

Keywords

Citrus;
Coccidiosis Eimeria spp.;
Grapefruit peel extract;
Naringenin

: Corresponding author.

Antiparasitic activity, histopathology and physiology of Colossoma macropomum (tambaqui) exposed to the essential oil of Lippia sidoides (Verbenaceae)

Veterinary Parasitology

Available online 16 December 2016

In Press, Accepted Manuscript — Note to users

Research paper

^a Programa de Pós-Graduação em Biodiversidade Tropical (PPGBIO), Universidade Federal do Amapá (UNIFAP), Macapá, AP, Brazil
^b Embrapa Amapá, Macapá, AP, Brazil
^c Programa de Pós-Graduação em Recursos Aquáticos Continentais Amazônicos (PPG-RACAM), Universidade do Oeste do Pará (UFOPA), Santarém, PA, Brazil
^d Embrapa Amazônia Ocidental, Manaus, AM, Brazil
^e Programa de Pós-Graduação em Aquicultura, Instituto Nacional de Pesquisas da Amazônia (INPA), Manaus, AM, Brazil

Received 10 August 2016, Revised 1 December 2016, Accepted 16 December 2016, Available online 16 December 2016

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http://dx.doi.org/10.1016/j.vetpar.2016.12.012

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Highlights

•: Anti-helminthic activity of the essential oil of Lippia sidoides against monogeneans of Colossoma macropomum is dose-dependent.
•: Lippia sidoides killed the parasites after 10 min in the in vitro trial.
•: Parasites exposed to water died after 8 h in the in vitro immobilization test.
•: Lippia sidoides causes anesthetic effect in fish and gills histopathological alterations.

Abstract

In vivo and in vitro antiparasitic activity of the essential oil of Lippia sidoides and blood and histological alterations were assessed in Colossoma macropomum (tambaqui). Essential oil concentrations of 10, 20, 40, 80, 160 and 320 mg/L were assayed in vitro against monogenoideans Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri from fish gills. Lippia sidoides essential oil concentrations of 320 and 160 mg/L were 100% effective against monogenoideans in 10 min and 1 h of exposure, respectively. However, the effectiveness of 100% concentrations of 80 mg/L and 40 mg/L occurred in 3 and 6 h, respectively. In the in vivo tests, juvenile fish were submitted to 60 min of baths with 10 mg/L and 15 min of baths with 20 mg/L of the essential oil of L. sidoides. These therapeutic baths were not efficient against Ichthyophthirius multifiliis, and monogenoideans present in the gills of C. macropomum. In addition, 10 and 20 mg/L of the essential oil of L. sidoides caused an anesthetic effect on the fish and did not influence total glucose and protein plasma levels; however, it decreased the number of total erythrocytes in fish exposed to the higher concentration of this essential oil. Severe alterations and irreversible damage were observed in the fish gills just after L. sidoides essential oil baths and after 24 h of recovery. The most recurrent lesions found were hyperplasia and fusion of the lamellar epithelium, vasodilation, detachment of the gill epithelium and lamellar aneurism, epithelial breakdown with hemorrhage, congestion, edema and necrosis, proliferation of the mucous cells and chloride cells and lamellar hypertrophy. Therefore, since the essential oil of L. sidoides has in vitro antiparasitic activity and low concentrations of it have shown toxic effects, the bioactive potential of its main chemical components should be investigated, as well as more efficient forms of its administration in therapeutic baths in order to eliminate fish parasites.

Keywords

Monogenoidea;
Parasites;
Medicinal plant;
Blood;
Tambaqui

: Corresponding author at: Embrapa Amapá, Rodovia Juscelino Kubitschek, km 5, 2600, 68903-419, Macapá, AP, Brazil.

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Accepted manuscripts are Articles in Press

The in vitro anthelmintic properties of browse species against Haemonchus contortus is determined by the polyphenol content and composition

Veterinary Parasitology

Available online 31 December 2016

In Press, Accepted Manuscript — Note to users

Research paper

^a Wageningen University & Research, Department of Animal Sciences, Animal Nutrition Group, PO Box 338, 6700 AH, Wageningen, the Netherlands
^b Department of Animal, Rangeland and Wildlife Sciences, Mekelle University, PO Box 231, Mekelle, Ethiopia
^c UMR 1225 IHAP INRA/ENVT, 23 Chemin des Capelles, Toulouse F-31076, France
^d Université de Toulouse, INP-ENVT, 23 Chemin des Capelles, Toulouse F-31076, France
^e Laboratory of Organic Chemistry and Chemical Biology, Department of Chemistry, University of Turku, FI-20014, Turku, Finland
^f Department of Animal Health, Utrecht University, PO Box 80163, 3508 TD, Utrecht, the Netherlands

Received 12 October 2016, Revised 23 December 2016, Accepted 29 December 2016, Available online 31 December 2016

http://dx.doi.org/10.1016/j.vetpar.2016.12.020

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Highlights

•: The browse plant species exhibit in vitro anthelmintic properties against Haemonchus contortus.
•: Proanthocyanidin concentration is not necessarily related to anthelmintic property.
•: Polyphenols other than tannins exert anthelmintic activity in the browse.
•: In some tannin containing browse, anthelmintic properties are mainly due to non-phenolic compounds.

Abstract

The aims of the present study were to (a) evaluate the anthelmintic activity of 10 East African browse extracts, (b) examine their role in inhibition of Haemonchus contortus larval exsheathment, (c) establish relationship between inhibition of larval exsheathment and browse extract polyphenol composition. Acetone/water (70/30%) extracts of air dried leaves of Acacia etbaica, Cadaba farinosa, Capparis tomentosa, Dichrostachys cinerea, Dodonaea angustifolia, Euclea racemosa, Maerua angolensis, Maytenus senegalensis, Rhus natalensis and Senna singueana were used. The larval exsheathment inhibition assay (LEIA) was applied using H. contortus third stage larvae (L₃) and extract concentrations of 0, 150, 300, 600, 1200 μg/ml phosphate buffered saline (PBS). Data were analysed using the PROC MIXED procedure of SAS. Polyvinylpolypyrrolidone (PVPP) was used to evaluate whether polyphenols were involved in L₃ exsheathment inhibition. All browse extracts significantly (P ≤ 0.0001) inhibited larval exsheathment in a dose dependent manner. The dose required to inhibit 50% of the larvae (EC₅₀) was highest in C. farinosa and lowest in E. racemosa and M. senegalensis. Significant differences (P < 0.001) between the control and PVPP treated A. etbaica, C. tomentosa, M. angolensis, R. natalensis and D. cinerea indicates that larval inhibition was largely due to non-phenol compounds. For E. racemosa, M. senegalensis, D. angustifolia and S. singueana, PVPP treatment reversed inhibition activity and in these extracts, inhibition was mostly attributable to tannin and other polyphenols (kaempferol, quercetin and myricetin based aglycones and glycosides). Overall, browse extracts have anthelmintic property against H. contortus and larval inhibition resulting from the presence of phenolic and non-phenolic compounds.

Keywords

Proanthocyanidins;
larvae;
exsheathment;
Haemonchus contortus

: Corresponding author at: Wageningen University & Research, Department of Animal Sciences, Animal Nutrition Group, 6708 WD, Wageningen, the Netherlands.

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Accepted manuscripts are Articles in Press

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