RESUMO
The present study aimed to evaluate the effect of nanoemulsions using combined synthetic anthelmintics, thiabendazole (TBZ), levamisole (LEV), and ivermectin (IVM), with carvacryl acetate (CA) against Haemonchus contortus, and also tested the presence and absence of alginate (ALG). The anthelmintic effect of the CA/TBZ nanoemulsion was evaluated in the egg hatch test (EHT). The effects of CA/IVM and CA/LEV nanoemulsions were evaluated in the larval development test (LDT). The emulsions CA/TBZ/ALG and CA/TBZ showed a multimodal profile, with most particles on the nanometric scale. The encapsulation efficiency in CA/TBZ/ALG was 80.25%, and that in CA/LEV/ALG was 89.73%. In the EHT, CA/TBZ and CA/TBZ/ALG showed mean combination indices (CIs) of 0.55 and 0.36, respectively, demonstrating synergism in both. In LDT, CA/IVM had an average CI of 0.75, and CA/LEV and CA/LEV/ALG showed CI values of 0.4 and 0.93, respectively. It was concluded that CA/TBZ showed a synergistic interaction, and CA/TBZ/ALG showed an enhanced effect. In addition, the matrix brought stability to the product, encouraging its improvement to obtain higher efficacy.
RESUMO
The aim of the present study was to evaluate the effects of the essential oils of Lippia alba chemotypes carvone and citral on H. contortus. Chemical characterization was performed by means of gas chromatography coupled to mass spectrometry (GC/MS). The anthelmintic effects of the essential oils were assessed through the egg hatch test (EHT) and the adult worm motility test (AWMT) using a multidrug-resistant H. contortus Kokstad isolate. Confocal laser scanning microscopy (CLSM) of eggs and adults of H. contortus and scanning electron microscopy (SEM) of adults were performed after treatment with oils for qualitative observations of their effects. The carvone chemotype of L. alba (LaCV) presented 70% carvone, and the citral chemotype of L. alba (LaCT) presented 29.4% geranial and 20.4% neral, respectively. In the EHT, the EC50 values of LaCV and LaCT were 0.2 and 0.3 mg/mL, respectively. In AWMT, after 12 h of exposure to 2 mg/mL LaCV and 2 mg/mL LaCT, 100% of adult nematodes were immobile. CLSM showed changes in larval motility inside the egg caused by LaCV, while LaCT promoted changes in larval formation. In adults exposed to both chemotypes, alterations in the anterior portion of the oesophagus were observed. In SEM, morphological changes were observed in the buccal capsule and in the medial portion of H. contortus adults. It is concluded that the two essential oils of L. alba, the chemotypes carvone and citral, caused morphological changes and inhibited the hatching of eggs and the motility of adult H. contortus nematodes.
Assuntos
Anti-Helmínticos , Haemonchus , Lippia , Óleos Voláteis , Animais , Óleos Voláteis/farmacologia , Lippia/química , Anti-Helmínticos/farmacologia , LarvaRESUMO
This work aimed to evaluate the in vitro anthelmintic effect of carvone nanoemulsions on Haemonchus contortus. Three R-carvone nanoemulsions were prepared: uncoated R-carvone nanoemulsions homogenized in a sonicator (UNAlg-son) and homogenized in an ultrahomogenizer (UNAlg-ultra) and sodium alginate-coated R-carvone (CNAlg-ultra). The physicochemical characterizations of the nanoemulsions were carried out. The anthelmintic activity was evaluated using egg hatch test (EHT), larval development test (LDT) and adult worm motility test (AWMT). Changes in cuticle induced in adult H. contortus were evaluated by scanning electron microscopy (SEM). The results were subjected to analysis of variance and compared using the Tukey test (P < 0.05). The effective concentration to inhibit 50% (EC50) of egg hatching and larval development was calculated. The particle sizes were 281.1 nm (UNAlg-son), 152.7 nm (UNAlg-ultra) and 557.8 nm (CNAlg-ultra), and the zeta potentials were −15 mV (UNAlg-son), −10.8 mV (UNAlg-ultra) and −24.2 mV (CNAlg-ultra). The encapsulation efficiency was 99.84 ± 0.01%. SEM of the nanoemulsions showed an increase in size. In EHT, the EC50 values of UNAlg-son, UNAlg-ultra and CNAlg-ultra were 0.19, 0.02 and 0.17 mg mL−1, respectively. In LDT, they were 0.29, 0.31 and 0.95 mg mL−1 for UNAlg-son, UNAlg-ultra and CNAlg-ultra, respectively. The adult motility inhibition was 100% after 12 h of exposure to UNAlg-ultra and CNAlg-ultra, while for UNAlg-son, it was 79.16%. SEM showed changes in the buccal capsule and cuticular damage. It was concluded that R-carvone nanoemulsions showed antiparasitic action demonstrating promise for the control of infections caused by gastrointestinal nematodes in small ruminants.
Assuntos
Anti-Helmínticos , Hemoncose , Haemonchus , Animais , Hemoncose/tratamento farmacológico , Hemoncose/veterinária , Hemoncose/parasitologia , Anti-Helmínticos/farmacologia , Anti-Helmínticos/uso terapêutico , Monoterpenos Cicloexânicos , Larva , Extratos Vegetais/farmacologiaRESUMO
Productivity in small ruminants can be compromised due to lice infestations. This pest infestation is controlled by the administration of insecticides, and their excessive use causes resistance. New control alternatives are being investiga-ted, especially the use of phytotherapeutics. Research using the oil from Carapa guianensis (andiroba) has shown insecticidal action on feline and goat lice specimens. The present study aimed to evaluate the efficacy of C. guianensis seed oil, in vitro, on Damalinia (Bovicola) ovis.Nine hundred specimens of D. (Bovicola) ovis were randomly assigned into groups and were subjected to different treatments: (G1) 1,000 to 15.6 mg/mL of C. guianensis oil; (G2) 3% Tween 80 (negative control); and (G3) amitraz 1.25 g/mL (positive control). Data presentation was expressed as mean efficacy ± standard deviation using the statistical program SPSS version 23.0 and statistical differences of the experimental groups were obtained by Kruskal-Wallis and Friedman. Probit Regression was performed to obtain CI50 and CI90, with a significance level of p < 0.05. The oil with a concentration of 1,000 mg/mL had an efficacy of 94% after 24 hours of exposure, differing statistically from the positive and negative control (p < 0.05). The IC50 and IC90 values were 439.21 mg/mL and 807.09 mg/mL, respectively. The action of the oil on the specimens was independent whether it was an adult female, male, or nymph. It is concluded that C. guianensis oil showed in vitro insecticidal effect against D. (Bovicola) ovis.(AU)
A produtividade em pequenos ruminantes pode ser comprometida devido as infestações por piolhos. O controle dessa infestação é realizado com a administração de inseticidas e seu uso excessivo causam resistências. Novas alternativas de controle estão sendo estudadas, destacando-se o uso de fitoterápicos. Pesquisas realizadas usando o óleo da Carapa guianensis(andiroba) mostraram ação inseticida sobre espécimes de piolhos de felinos e caprinos. O presente estudo teve como objetivo avaliar a eficácia do óleo da semente de C. guianensis, in vitro, sobre Damalinia (Bovicola) ovis. 900 exemplares de D. (Bovicola) ovis foram distribuídos aleatoriamente em grupos e submetidos a diferentes tratamentos: (G1) 1.000 a 15,6 mg/ml de óleo de C. guianensis; (G2) Tween 80 a 3% (controle negativo), e: (G3) Amitraz 1,25 g/mL (controle positivo). A apresentação dos dados foi expressa em eficácia média ± desvio padrão usando o programa estatístico SPSS versão 23.0 e diferenças estatísticas dos grupos experimentais foram obtidas por Kruskal-Wallis e Friedman. Para obtenção da CI50 e CI90 foi realizada Regressão de Probit, com nível de significância de p< 0,05. O óleo com concentração de 1.000 mg/ml teve uma eficácia de 94%, após 24 horas de exposição, diferindo estatisticamente do controle positivo e negativo (p< 0,05). Os valores da CI50 e CI90 foram de 439,21 mg/mL e 807,09 mg/mL, respectivamente. A ação do óleo sobre os espécimes foi independente se era adulto fêmea, macho ou ninfa. Conclui-se que o óleo de C. guianensis apresentou efeito inseticida in vitro contra D. (Bovicola) ovis.(AU)
Assuntos
Animais , Ovinos/parasitologia , Infestações por Piolhos , Técnicas In Vitro , Meliaceae/química , Iscnóceros/químicaRESUMO
Productivity in small ruminants can be compromised due to lice infestations. This pest infestation is controlled by the administration of insecticides, and their excessive use causes resistance. New control alternatives are being investiga-ted, especially the use of phytotherapeutics. Research using the oil from Carapa guianensis (andiroba) has shown insecticidal action on feline and goat lice specimens. The present study aimed to evaluate the efficacy of C. guianensis seed oil, in vitro, on Damalinia (Bovicola) ovis.Nine hundred specimens of D. (Bovicola) ovis were randomly assigned into groups and were subjected to different treatments: (G1) 1,000 to 15.6 mg/mL of C. guianensis oil; (G2) 3% Tween 80 (negative control); and (G3) amitraz 1.25 g/mL (positive control). Data presentation was expressed as mean efficacy ± standard deviation using the statistical program SPSS version 23.0 and statistical differences of the experimental groups were obtained by Kruskal-Wallis and Friedman. Probit Regression was performed to obtain CI50 and CI90, with a significance level of p < 0.05. The oil with a concentration of 1,000 mg/mL had an efficacy of 94% after 24 hours of exposure, differing statistically from the positive and negative control (p < 0.05). The IC50 and IC90 values were 439.21 mg/mL and 807.09 mg/mL, respectively. The action of the oil on the specimens was independent whether it was an adult female, male, or nymph. It is concluded that C. guianensis oil showed in vitro insecticidal effect against D. (Bovicola) ovis.
A produtividade em pequenos ruminantes pode ser comprometida devido as infestações por piolhos. O controle dessa infestação é realizado com a administração de inseticidas e seu uso excessivo causam resistências. Novas alternativas de controle estão sendo estudadas, destacando-se o uso de fitoterápicos. Pesquisas realizadas usando o óleo da Carapa guianensis(andiroba) mostraram ação inseticida sobre espécimes de piolhos de felinos e caprinos. O presente estudo teve como objetivo avaliar a eficácia do óleo da semente de C. guianensis, in vitro, sobre Damalinia (Bovicola) ovis. 900 exemplares de D. (Bovicola) ovis foram distribuídos aleatoriamente em grupos e submetidos a diferentes tratamentos: (G1) 1.000 a 15,6 mg/ml de óleo de C. guianensis; (G2) Tween 80 a 3% (controle negativo), e: (G3) Amitraz 1,25 g/mL (controle positivo). A apresentação dos dados foi expressa em eficácia média ± desvio padrão usando o programa estatístico SPSS versão 23.0 e diferenças estatísticas dos grupos experimentais foram obtidas por Kruskal-Wallis e Friedman. Para obtenção da CI50 e CI90 foi realizada Regressão de Probit, com nível de significância de p< 0,05. O óleo com concentração de 1.000 mg/ml teve uma eficácia de 94%, após 24 horas de exposição, diferindo estatisticamente do controle positivo e negativo (p< 0,05). Os valores da CI50 e CI90 foram de 439,21 mg/mL e 807,09 mg/mL, respectivamente. A ação do óleo sobre os espécimes foi independente se era adulto fêmea, macho ou ninfa. Conclui-se que o óleo de C. guianensis apresentou efeito inseticida in vitro contra D. (Bovicola) ovis.
Assuntos
Animais , Infestações por Piolhos , Iscnóceros/química , Meliaceae/química , Ovinos/parasitologia , Técnicas In VitroRESUMO
The aim of this study was to characterize the anthelmintic resistance (AR) of a sheep gastrointestinal nematode population, named Caucaia, from northeastern Brazil. Phenotypic tests performed were: egg hatch (EHT), larval development (LDT) and fecal egg count reduction (FECRT). Benzimidazoles (BZs) genotypic evaluation was by frequency of single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A, and for levamisole (LEV), by frequency of resistance alleles of Hco-acr-8 gene. The primers were designed specifically for Haemonchus contortus. Effective concentrations 50% (EC50) for BZs (EHT), and for macrocyclic lactones (MLs) and LEV (LDT) were 1.02 µg/mL, 1.81 ng/mL and 0.04 µg/mL, respectively. Resistance ratios for MLs and LEV were 0.91 and 3.07, respectively. FECRT efficacies of BZs, MLs, monepantel (MPTL) and LEV were 52.4; 87.0; 94.5 and 99.6%, respectively. qPCR for BZs demonstrated resistance allele frequencies of 0%, 26.24% and 69.08% for SNPs E198A, F200Y and F167Y, respectively. For LEV, 54.37% of resistance alleles were found. There was agreement between EHT, FECRT and qPCR for BZs, and agreement between LDT and qPCR for LEV. Thus, based on higher sensitivity of qPCR, and phenotypic evaluation, the Caucaia population was considered resistant to BZs, MLs, LEV and suspect for MPTL.
Assuntos
Anti-Helmínticos , Haemonchus , Nematoides , Doenças dos Ovinos , Animais , Anti-Helmínticos/farmacologia , Anti-Helmínticos/uso terapêutico , Brasil , Resistência a Medicamentos/genética , Fezes , Contagem de Ovos de Parasitas/veterinária , Ovinos , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/tratamento farmacológicoRESUMO
O objetivo deste trabalho é avaliar a eficácia inseticida in vitro do óleo essencial de Lippia gracilis sobre Bovicolaovis. Os espécimes (n = 900) de B.ovis foram coletados de ovinos naturalmente infestados, distribuídos aleatoriamente em grupos, e submetidos aos diferentes tratamentos: (G1) 1.000 a 15,6 mg/ml óleo essencial de Lippia gracilis; (G2) Tween 80 a 3% (controle negativo), e: (G3) Amitraz 1,25 g/mL (controle positivo). Realizou-se 4 réplicas (25 piolhos por réplica) para cada tratamento. A viabilidade dos piolhos foi avaliada nos tempos de 1, 3, 6, 24, 48 e 72 h. Os dados foram expressos em eficácia média ± desvio padrão (SPSS versão 23.0) e diferenças estatísticas dos grupos experimentais foram obtidas por Kruskal-Wallis e Friedman. Para obtenção da CI50e CI90 foi realizada Regressão de Probit, com nível de significância de p< 0,05. O óleo essencial de L. gracilis nas concentrações de 100 a 62,5 mg/mL apresentaram eficácia de 100%, 1 h pós-exposição. Os valores de CI50 e CI90 foram 18,1 mg/mL e 44,9 mg/mL, respectivamente. Dessa forma, concluímos que o óleo essencial de L. gracilis apresentou eficácia sobre B.ovis in vitro.
The objective of this work is to evaluate the in vitro insecticidal efficacy of the essential oil of Lippia gracilis against Bovicolaovis. The specimens (n = 900) of B.ovis were collected from naturally infested sheep, randomly distributed in groups, and subjected to different treatments: (G1) 1,000 to 15.6 mg/ml essential oil of Lippia gracilis; (G2) 3% Tween 80 (negative control), and: (G3) Amitraz 1.25 g/mL (positive control). 4 replicates (25 lice per replica) were performed for each treatment. Louse viability was assessed at 1, 3, 6, 24, 48 and 72 h. The data were expressed as mean efficacy ± standard deviation (SPSS version 23.0) and statistical differences in the experimental groups were obtained by Kruskal-Wallis and Friedman. Probit regression was performed to obtain the IC50 and CI90, with a significance level of p <0.05. The essential oil of L. gracilis at concentrations of 100 to 62.5 mg/mL showed 100% efficacy, 1 h post-exposure. The IC50 and CI90 values were 18.1 mg/mL and 44.9 mg/mL, respectively. Thus, we conclude that L. gracilis essential oil is effective on B. ovis.
Assuntos
Animais , Infestações por Piolhos/veterinária , Óleos Voláteis/uso terapêutico , Lippia/toxicidade , Inseticidas , Ovinos/parasitologia , Medicamento FitoterápicoRESUMO
O objetivo deste trabalho é avaliar a eficácia inseticida in vitro do óleo essencial de Lippia gracilis sobre Bovicolaovis. Os espécimes (n = 900) de B.ovis foram coletados de ovinos naturalmente infestados, distribuídos aleatoriamente em grupos, e submetidos aos diferentes tratamentos: (G1) 1.000 a 15,6 mg/ml óleo essencial de Lippia gracilis; (G2) Tween 80 a 3% (controle negativo), e: (G3) Amitraz 1,25 g/mL (controle positivo). Realizou-se 4 réplicas (25 piolhos por réplica) para cada tratamento. A viabilidade dos piolhos foi avaliada nos tempos de 1, 3, 6, 24, 48 e 72 h. Os dados foram expressos em eficácia média ± desvio padrão (SPSS versão 23.0) e diferenças estatísticas dos grupos experimentais foram obtidas por Kruskal-Wallis e Friedman. Para obtenção da CI50e CI90 foi realizada Regressão de Probit, com nível de significância de p< 0,05. O óleo essencial de L. gracilis nas concentrações de 100 a 62,5 mg/mL apresentaram eficácia de 100%, 1 h pós-exposição. Os valores de CI50 e CI90 foram 18,1 mg/mL e 44,9 mg/mL, respectivamente. Dessa forma, concluímos que o óleo essencial de L. gracilis apresentou eficácia sobre B.ovis in vitro.
The objective of this work is to evaluate the in vitro insecticidal efficacy of the essential oil of Lippia gracilis against Bovicolaovis. The specimens (n = 900) of B.ovis were collected from naturally infested sheep, randomly distributed in groups, and subjected to different treatments: (G1) 1,000 to 15.6 mg/ml essential oil of Lippia gracilis; (G2) 3% Tween 80 (negative control), and: (G3) Amitraz 1.25 g/mL (positive control). 4 replicates (25 lice per replica) were performed for each treatment. Louse viability was assessed at 1, 3, 6, 24, 48 and 72 h. The data were expressed as mean efficacy ± standard deviation (SPSS version 23.0) and statistical differences in the experimental groups were obtained by Kruskal-Wallis and Friedman. Probit regression was performed to obtain the IC50 and CI90, with a significance level of p <0.05. The essential oil of L. gracilis at concentrations of 100 to 62.5 mg/mL showed 100% efficacy, 1 h post-exposure. The IC50 and CI90 values were 18.1 mg/mL and 44.9 mg/mL, respectively. Thus, we conclude that L. gracilis essential oil is effective on B. ovis.
Assuntos
Animais , Compostos Fitoquímicos , Inseticidas/análise , Ruminantes/fisiologia , Óleos Voláteis/toxicidade , Lippia/toxicidade , Técnicas In VitroRESUMO
O objetivo deste trabalho é avaliar a eficácia inseticida in vitro do óleo essencial de Lippia gracilis sobre Bovicolaovis. Os espécimes (n = 900) de B.ovis foram coletados de ovinos naturalmente infestados, distribuídos aleatoriamente em grupos, e submetidos aos diferentes tratamentos: (G1) 1.000 a 15,6 mg/ml óleo essencial de Lippia gracilis; (G2) Tween 80 a 3% (controle negativo), e: (G3) Amitraz 1,25 g/mL (controle positivo). Realizou-se 4 réplicas (25 piolhos por réplica) para cada tratamento. A viabilidade dos piolhos foi avaliada nos tempos de 1, 3, 6, 24, 48 e 72 h. Os dados foram expressos em eficácia média ± desvio padrão (SPSS versão 23.0) e diferenças estatísticas dos grupos experimentais foram obtidas por Kruskal-Wallis e Friedman. Para obtenção da CI50e CI90 foi realizada Regressão de Probit, com nível de significância de p< 0,05. O óleo essencial de L. gracilis nas concentrações de 100 a 62,5 mg/mL apresentaram eficácia de 100%, 1 h pós-exposição. Os valores de CI50 e CI90 foram 18,1 mg/mL e 44,9 mg/mL, respectivamente. Dessa forma, concluímos que o óleo essencial de L. gracilis apresentou eficácia sobre B.ovis in vitro.(AU)
The objective of this work is to evaluate the in vitro insecticidal efficacy of the essential oil of Lippia gracilis against Bovicolaovis. The specimens (n = 900) of B.ovis were collected from naturally infested sheep, randomly distributed in groups, and subjected to different treatments: (G1) 1,000 to 15.6 mg/ml essential oil of Lippia gracilis; (G2) 3% Tween 80 (negative control), and: (G3) Amitraz 1.25 g/mL (positive control). 4 replicates (25 lice per replica) were performed for each treatment. Louse viability was assessed at 1, 3, 6, 24, 48 and 72 h. The data were expressed as mean efficacy ± standard deviation (SPSS version 23.0) and statistical differences in the experimental groups were obtained by Kruskal-Wallis and Friedman. Probit regression was performed to obtain the IC50 and CI90, with a significance level of p <0.05. The essential oil of L. gracilis at concentrations of 100 to 62.5 mg/mL showed 100% efficacy, 1 h post-exposure. The IC50 and CI90 values were 18.1 mg/mL and 44.9 mg/mL, respectively. Thus, we conclude that L. gracilis essential oil is effective on B. ovis.(AU)
Assuntos
Animais , Ruminantes/fisiologia , Óleos Voláteis/toxicidade , Inseticidas/análise , Compostos Fitoquímicos , Técnicas In Vitro , Lippia/toxicidadeRESUMO
The objective of this study was to evaluate the efficacy of carvacryl acetate (CVA) and nanoencapsulated CVA (nCVA) on gastrointestinal nematodes of sheep. The CVA was nanoencapsulated with chitosan/gum arabic and the efficacy of nanoencapsulation (EE), yield, zeta potential, nanoparticle morphology and release kinetics at pH 3 and 8 were analyzed. Acute and subchronic toxicity were evaluated in rodents and reduction of egg counts in the faeces (FECRT) of sheep. The sheep were divided into four groups (n = 10): G1, 250 mg/kg CVA; G2, 250 mg/kg nCVA; G3, polymer matrix and G4: 2.5 mg/kg monepantel. EE and nCVA yield were 65% and 57%, respectively. The morphology of the nanoparticles was spherical, size (810.6±286.7 nm), zeta potential in pH 3.2 (+18.3 mV) and the 50% release of CVA at pHs 3 and 8 occurred at 200 and 10 h, respectively. nCVA showed LD50 of 2,609 mg/kg. CVA, nCVA and monepantel reduced the number of eggs per gram of faeces (epg) by 57.7%, 51.1% and 97.7%, respectively. The epg of sheep treated with CVA and nCVA did not differ from the negative control (P>0.05). Nanoencapsulation reduced the toxicity of CVA; however, nCVA and CVA presented similar results in the FECRT.
Assuntos
Anti-Helmínticos/administração & dosagem , Trato Gastrointestinal/parasitologia , Monoterpenos/administração & dosagem , Nanocápsulas/administração & dosagem , Nematoides/efeitos dos fármacos , Infecções por Nematoides/tratamento farmacológico , Doenças dos Ovinos/tratamento farmacológico , Animais , Anti-Helmínticos/toxicidade , Fezes/parasitologia , Feminino , Camundongos , Monoterpenos/toxicidade , Nanocápsulas/toxicidade , Nematoides/classificação , Nematoides/isolamento & purificação , Infecções por Nematoides/parasitologia , Contagem de Ovos de Parasitas , Testes de Sensibilidade Parasitária , Reação em Cadeia da Polimerase em Tempo Real , Roedores , Ovinos , Doenças dos Ovinos/parasitologia , Testes de ToxicidadeRESUMO
Abstract The objective of this study was to evaluate the efficacy of carvacryl acetate (CVA) and nanoencapsulated CVA (nCVA) on gastrointestinal nematodes of sheep. The CVA was nanoencapsulated with chitosan/gum arabic and the efficacy of nanoencapsulation (EE), yield, zeta potential, nanoparticle morphology and release kinetics at pH 3 and 8 were analyzed. Acute and subchronic toxicity were evaluated in rodents and reduction of egg counts in the faeces (FECRT) of sheep. The sheep were divided into four groups (n = 10): G1, 250 mg/kg CVA; G2, 250 mg/kg nCVA; G3, polymer matrix and G4: 2.5 mg/kg monepantel. EE and nCVA yield were 65% and 57%, respectively. The morphology of the nanoparticles was spherical, size (810.6±286.7 nm), zeta potential in pH 3.2 (+18.3 mV) and the 50% release of CVA at pHs 3 and 8 occurred at 200 and 10 h, respectively. nCVA showed LD50 of 2,609 mg/kg. CVA, nCVA and monepantel reduced the number of eggs per gram of faeces (epg) by 57.7%, 51.1% and 97.7%, respectively. The epg of sheep treated with CVA and nCVA did not differ from the negative control (P>0.05). Nanoencapsulation reduced the toxicity of CVA; however, nCVA and CVA presented similar results in the FECRT.
Resumo O objetivo deste trabalho foi avaliar a eficácia do acetato de carvacrila (ACV) e do ACV nanoencapsulado (nACV) sobre nematóides gastrintestinais de ovinos. O ACV foi nanoencapsulado com quitosana/goma arábica e foi analisada a eficácia de nanoencapsulamento (EE), o rendimento, potencial zeta, morfologia das nanopartículas e cinética de liberação em pH 3 e 8. Foram avaliadas as toxicidades aguda e subcrônica em roedores e a redução da contagem de ovos nas fezes (RCOF) de ovinos. Os ovinos foram divididos em quatro grupos (n = 10): G1, 250 mg/kg ACV; G2, 250 mg/kg de nACV; G3, matriz polimérica e G4: 2,5 mg/kg de monepantel. A EE e o rendimento de nACV foram de 65% e 57%, respectivamente. A morfologia das nanopartículas foi esférica, tamanho (810,6±286,7 nm), potencial zeta no pH 3,2 (+18,3 mV) e a liberação de 50% de CVA nos pHs 3 e 8 ocorreu às 200 e 10 h, respectivamente. nACV apresentou DL50 de 2.609 mg/kg. ACV, nACV e o monepantel reduziram a contagem de ovos por grama de fezes (opg) em 57,7%, 51,1% e 97,7%, respectivamente. A contagem de opg de ovelhas tratadas com ACV e nCVA não diferiu do controle negativo (P>0,05). O nanoencapsulamento reduziu a toxicidade do AVC; no entanto, nACV e ACV apresentaram resultados semelhantes na RCOF.
Assuntos
Animais , Feminino , Camundongos , Ratos , Doenças dos Ovinos/parasitologia , Monoterpenos/farmacologia , Trato Gastrointestinal/parasitologia , Nanocápsulas/administração & dosagem , Anti-Helmínticos/farmacologia , Infecções por Nematoides/veterinária , Contagem de Ovos de Parasitas , Doenças dos Ovinos/tratamento farmacológico , Benzimidazóis/farmacologia , Resistência a Medicamentos/efeitos dos fármacos , Ovinos/parasitologia , Levamisol/farmacologia , Ratos Wistar/sangue , Testes de Toxicidade , Testes de Sensibilidade Parasitária , Monoterpenos/toxicidade , Monoterpenos/uso terapêutico , Nanocápsulas/toxicidade , Nanocápsulas/uso terapêutico , Reação em Cadeia da Polimerase em Tempo Real , Hemoncose/tratamento farmacológico , Haemonchus/isolamento & purificação , Haemonchus/efeitos dos fármacos , Helmintíase Animal/tratamento farmacológico , Anti-Helmínticos/toxicidade , Anti-Helmínticos/uso terapêutico , Camundongos , Infecções por Nematoides/tratamento farmacológicoRESUMO
This study aimed to evaluate the anthelmintic and ultrastructural effects of Calotropis procera latex on Haemonchus contortus. C. procera latex was twice centrifuged at 10,000×g and dialyzed to obtain a fraction rich in proteins, named LP (latex protein), and at 3,000 rpm to obtain a fraction rich in secondary metabolites, named LNP (latex non-protein). Specimens of H. contortus exposed to LNP, LP and PBS in the Adult Worm Motility Test (AWMT) were submitted to scanning (SEM) and transmission (TEM) electron microscopy to verify changes in their ultrastructure. Phytochemical tests in the LNP indicated the presence of phenols, steroids, alkaloids and cardenolides. High-Performance Liquid Chromatography (HPLC) characterized the presence of the compounds gallic acid and quercetin in the LNP. The protein content in the LP was 43.1 ± 1.1 mg/mL and 7.7 ± 0.3 mg/mL in LNP. In AWMT, LNP and LP inhibited the motility of 100% of the nematodes, with LNP being more effective than LP and ivermectin more effective than both (p <0.05). Cuticle changes were observed by SEM and TEM in nematodes treated with LP and LNP. Calotropis procera latex has anthelmintic effects against H. contortus, causing damage to its cuticle and other alterations in its ultrastructure.(AU)
Este estudo objetivou avaliar os efeitos anti-helmínticos e ultraestruturais do látex de Calotropis procera sobre Haemonchus contortus. Látex de C. procera foi centrifugado duas vezes à a 10.000xg e dialisado para obter uma fração rica em proteínas, denominada proteínas do látex (LP). E centrifugado e centrifugado a 3.000 rpm, para obter uma fração rica em metabólitos secundários, denominada LNP (látex não proteico). Espécimes de H. contortus expostos à LNP, LP e PBS no Teste de Motilidade dos Nematoides Adultos (TMNA) foram submetidos a microscopia eletrônica de varredura (MEV) e de transmissão (MET), para verificar alterações em sua ultraestrutura. Testes fitoquímicos em LNP indicaram a presença de fenóis, esteroides, alcaloides e cardenolídeos. A presença dos compostos ácido gálico e quercetina em LNP foi caracterizada por Cromatografia Líquida de Alta Eficiência (CLAE). O conteúdo de proteínas em LP foi de 43,1 ± 1,1 mg/mL e de 7,7 ± 0,3 mg/mL em LNP. No TMNA, LNP e LP inibiram a motilidade de 100% dos nematoides, sendo LNP mais eficaz que LP, e a ivermectina mais eficaz que ambos (p <0,05). Alterações na cutícula de nematoides tratados com LP e LNP foram observadas por MEV e MET. O látex de C. procera apresenta efeito anti-helmíntico sobre H. contortus, causando danos à sua cutícula e outras alterações em sua ultraestrutura.(AU)
Assuntos
Animais , Calotropis/química , Haemonchus/química , Haemonchus/parasitologia , Anti-HelmínticosRESUMO
The objective of this study was to evaluate the efficacy of carvacryl acetate (CVA) and nanoencapsulated CVA (nCVA) on gastrointestinal nematodes of sheep. The CVA was nanoencapsulated with chitosan/gum arabic and the efficacy of nanoencapsulation (EE), yield, zeta potential, nanoparticle morphology and release kinetics at pH 3 and 8 were analyzed. Acute and subchronic toxicity were evaluated in rodents and reduction of egg counts in the faeces (FECRT) of sheep. The sheep were divided into four groups (n = 10): G1, 250 mg/kg CVA; G2, 250 mg/kg nCVA; G3, polymer matrix and G4: 2.5 mg/kg monepantel. EE and nCVA yield were 65% and 57%, respectively. The morphology of the nanoparticles was spherical, size (810.6±286.7 nm), zeta potential in pH 3.2 (+18.3 mV) and the 50% release of CVA at pHs 3 and 8 occurred at 200 and 10 h, respectively. nCVA showed LD50 of 2,609 mg/kg. CVA, nCVA and monepantel reduced the number of eggs per gram of faeces (epg) by 57.7%, 51.1% and 97.7%, respectively. The epg of sheep treated with CVA and nCVA did not differ from the negative control (P>0.05). Nanoencapsulation reduced the toxicity of CVA; however, nCVA and CVA presented similar results in the FECRT.(AU)
O objetivo deste trabalho foi avaliar a eficácia do acetato de carvacrila (ACV) e do ACV nanoencapsulado (nACV) sobre nematóides gastrintestinais de ovinos. O ACV foi nanoencapsulado com quitosana/goma arábica e foi analisada a eficácia de nanoencapsulamento (EE), o rendimento, potencial zeta, morfologia das nanopartículas e cinética de liberação em pH 3 e 8. Foram avaliadas as toxicidades aguda e subcrônica em roedores e a redução da contagem de ovos nas fezes (RCOF) de ovinos. Os ovinos foram divididos em quatro grupos (n = 10): G1, 250 mg/kg ACV; G2, 250 mg/kg de nACV; G3, matriz polimérica e G4: 2,5 mg/kg de monepantel. A EE e o rendimento de nACV foram de 65% e 57%, respectivamente. A morfologia das nanopartículas foi esférica, tamanho (810,6±286,7 nm), potencial zeta no pH 3,2 (+18,3 mV) e a liberação de 50% de CVA nos pHs 3 e 8 ocorreu às 200 e 10 h, respectivamente. nACV apresentou DL50 de 2.609 mg/kg. ACV, nACV e o monepantel reduziram a contagem de ovos por grama de fezes (opg) em 57,7%, 51,1% e 97,7%, respectivamente. A contagem de opg de ovelhas tratadas com ACV e nCVA não diferiu do controle negativo (P>0,05). O nanoencapsulamento reduziu a toxicidade do AVC; no entanto, nACV e ACV apresentaram resultados semelhantes na RCOF.(AU)
Assuntos
Animais , Ovinos/parasitologia , Roedores/parasitologia , Gastroenteropatias/parasitologia , AcetatosRESUMO
This study aimed to evaluate the anthelmintic activity of Eucalyptus citriodora essential oil and citronellal on sheep gastrointestinal nematodes. Essential oil composition was determined by gas chromatography mass spectrometry. The substances were evaluated in vitro using adult worm motility test (AWMT) and transmission electron microscopy (TEM). The acute toxicity test in mice and the fecal egg count reduction test (FECRT) in sheep were performed. Citronellal was confirmed as the essential oil major constituent (63.9%). According to the AWMT, 2 mg/mL of essential oil and citronellal completely inhibited Haemonchus contortus motility at 6 h post exposure. H. contortus exposed to essential oil and citronellal exhibited internal ultrastructural modifications. The lethal dose 50 values in mice were 5,000 and 2,609 mg/kg for essential oil and citronellal, respectively. E. citriodora essential oil reduced sheep epg at 14 days post treatment by 69.5% (P<0.05). No significant differences were observed in epg between the citronellal and negative control groups (P>0.05). The interaction between citronellal and other constituents in the essential oil may be relevant for its in vivo anthelmintic activity. Thus, E. citriodora essential oil and citronellal pharmacokinetic studies may help elucidate the anthelmintic activity of these compounds.
Assuntos
Anti-Helmínticos/farmacologia , Eucalyptus/química , Hemoncose/veterinária , Haemonchus/efeitos dos fármacos , Óleos Voláteis/farmacologia , Doenças dos Ovinos/parasitologia , Ovinos/parasitologia , Animais , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Hemoncose/parasitologia , Haemonchus/isolamento & purificação , Haemonchus/ultraestrutura , Camundongos , Microscopia Eletrônica de Transmissão , Óleos Voláteis/isolamento & purificação , Contagem de Ovos de Parasitas , Testes de Sensibilidade ParasitáriaRESUMO
Abstract This study aimed to evaluate the anthelmintic activity of Eucalyptus citriodora essential oil and citronellal on sheep gastrointestinal nematodes. Essential oil composition was determined by gas chromatography mass spectrometry. The substances were evaluated in vitro using adult worm motility test (AWMT) and transmission electron microscopy (TEM). The acute toxicity test in mice and the fecal egg count reduction test (FECRT) in sheep were performed. Citronellal was confirmed as the essential oil major constituent (63.9%). According to the AWMT, 2 mg/mL of essential oil and citronellal completely inhibited Haemonchus contortus motility at 6 h post exposure. H. contortus exposed to essential oil and citronellal exhibited internal ultrastructural modifications. The lethal dose 50 values in mice were 5,000 and 2,609 mg/kg for essential oil and citronellal, respectively. E. citriodora essential oil reduced sheep epg at 14 days post treatment by 69.5% (P<0.05). No significant differences were observed in epg between the citronellal and negative control groups (P>0.05). The interaction between citronellal and other constituents in the essential oil may be relevant for its in vivo anthelmintic activity. Thus, E. citriodora essential oil and citronellal pharmacokinetic studies may help elucidate the anthelmintic activity of these compounds.
Resumo Este trabalho objetivou avaliar a atividade anti-helmíntica do óleo essencial de Eucalyptus citriodora e citronelal sobre nematoides gastrintestinais de ovinos. A composição do óleo essencial foi determinada por cromatografia gasosa acoplada à espectrometria de massas. As substâncias foram avaliadas in vitro utilizando-se teste de motilidade de vermes adultos (AWMT) e microscopia eletrônica de transmissão (TEM). Teste de toxicidade aguda em camundongos e teste de redução da contagem de ovos fecais (FECRT) em ovinos foram realizados. Citronelal foi confirmado como componente majoritário do óleo essencial (63,9%). No AWMT, 2 mg/mL de óleo essencial e citronelal inibiram completamente a motilidade de H. contortus 6 h pós-exposição. H. contortus expostos ao óleo essencial e citronelal exibiram modificações ultraestruturais internas. Os valores da dose letal 50 em camundongos foram 5.000 e 2.609 mg/kg para óleo essencial e citronelal, respectivamente. Óleo essencial de E. citriodora reduziu opg de ovinos 14 dias pós-tratamento em 69,5% (P<0,05). Não houve diferença significativa de opg entre grupo controle negativo e citronelal (P>0,05). A interação entre citronelal e outros constituintes do óleo essencial pode ser relevante na atividade anti-helmíntica in vivo. Portanto, avaliação farmacocinética do óleo essencial de E. citriodora e citronelal pode auxiliar a elucidar a atividade anti-helmíntica desses compostos.
Assuntos
Animais , Feminino , Camundongos , Doenças dos Ovinos/parasitologia , Óleos Voláteis/farmacologia , Ovinos/parasitologia , Eucalyptus/química , Hemoncose/veterinária , Haemonchus/efeitos dos fármacos , Anti-Helmínticos/farmacologia , Contagem de Ovos de Parasitas , Óleos Voláteis/isolamento & purificação , Testes de Sensibilidade Parasitária , Microscopia Eletrônica de Transmissão , Hemoncose/parasitologia , Haemonchus/isolamento & purificação , Haemonchus/ultraestrutura , Cromatografia Gasosa-Espectrometria de MassasRESUMO
The anthelmintic resistance stimulated the search for strategies for controlling gastrointestinal nematodes, including the use of free essential oils or its nanoemulsion. This study evaluated the anthelmintic efficacy of Cymbopogon citratus essential oil (CcEO) and C. citratus essential oil nanoemulsion (CcEOn). Pysicochemical analyses were performed. The in vitro effect was determined using the egg hatch test (EHT) on Haemonchus contortus and in vivo effect was evaluated in sheep infected with gastrointestinal nematodes. The animals were treated with CcEO (500 mg/kg) or CcEOn (450 mg/kg) for the fecal egg count (FEC) and the determination of worm burden. The main component of CcEO was citral. The CcEO content in the nanoemulsion was 20% (v/v), and the mean particle size was 248 nm. In EHT, CcEO and CcEOn (1.25 mg/mL) inhibited larval hatching by 98.4 and 97.1%, respectively. Three animals treated with CcEO died whereas in the group treated with CcEOn one animal died. The FEC and total worm burden of the treated groups did not differ from the negative control (p>0.05). The CcEOn showed efficacy only on H. contortus (p<0.05). In conclusion, nanoencapsulation reduced toxicity and increased efficacy on H. contortus.
Assuntos
Anti-Helmínticos/farmacologia , Cymbopogon/química , Nematoides/efeitos dos fármacos , Óleos Voláteis/farmacologia , Extratos Vegetais/farmacologia , Doenças dos Ovinos/parasitologia , Animais , Fezes/parasitologia , Feminino , Masculino , Nematoides/classificação , Nematoides/isolamento & purificação , OvinosRESUMO
Abstract The anthelmintic resistance stimulated the search for strategies for controlling gastrointestinal nematodes, including the use of free essential oils or its nanoemulsion. This study evaluated the anthelmintic efficacy of Cymbopogon citratus essential oil (CcEO) and C. citratus essential oil nanoemulsion (CcEOn). Pysicochemical analyses were performed. The in vitro effect was determined using the egg hatch test (EHT) on Haemonchus contortus and in vivo effect was evaluated in sheep infected with gastrointestinal nematodes. The animals were treated with CcEO (500 mg/kg) or CcEOn (450 mg/kg) for the fecal egg count (FEC) and the determination of worm burden. The main component of CcEO was citral. The CcEO content in the nanoemulsion was 20% (v/v), and the mean particle size was 248 nm. In EHT, CcEO and CcEOn (1.25 mg/mL) inhibited larval hatching by 98.4 and 97.1%, respectively. Three animals treated with CcEO died whereas in the group treated with CcEOn one animal died. The FEC and total worm burden of the treated groups did not differ from the negative control (p>0.05). The CcEOn showed efficacy only on H. contortus (p<0.05). In conclusion, nanoencapsulation reduced toxicity and increased efficacy on H. contortus.
Resumo A resistência anti-helmíntica estimulou a busca por estratégias de controle de nematoides gastrintestinais, incluindo óleos essenciais livres ou em nanoemulsão. Este estudo avaliou a eficácia anti-helmíntica do óleo essencial de Cymbopogon citratus (OECc) e da nanoemulsão do óleo essencial de C. citratus (nOECc). Análises físico-químicas foram realizadas. O efeito in vitro foi determinado no teste de eclosão de ovos (TEO) sobre Haemonchus contortus e o efeito in vivo foi avaliado em ovinos infectados com nematoides gastrintestinais. Os animais foram tratados com OECc (500 mg/kg) ou nOECc (450 mg/kg) para determinação do número de ovos por grama de fezes (OPG) e carga parasitária. O principal constituinte do OECc foi citral. O conteúdo de OECc na nanoemulsão foi 20% e o tamanho médio de partícula foi 248 nm. No TEO, OECc e nOECc (1,25 mg/mL) inibiram 98,4 e 97,1% da eclosão larvar, respectivamente. Três animais tratados com o OECc morreram, enquanto um animal do grupo tratado com a nOECc morreu. O OPG e a carga parasitária total dos grupos tratados não diferiram do controle negativo (p>0.05). A nOECc apresentou eficácia somente sobre H. contortus (p<0.05). Como conclusão, o nanoencapsulamento reduziu a toxicidade e aumentou a eficácia sobre H. contortus.
Assuntos
Animais , Masculino , Feminino , Doenças dos Ovinos/parasitologia , Óleos Voláteis/farmacologia , Extratos Vegetais/farmacologia , Cymbopogon/química , Anti-Helmínticos/farmacologia , Nematoides/efeitos dos fármacos , Ovinos , Fezes/parasitologia , Nematoides/isolamento & purificação , Nematoides/classificaçãoRESUMO
The anthelmintic resistance stimulated the search for strategies for controlling gastrointestinal nematodes, including the use of free essential oils or its nanoemulsion. This study evaluated the anthelmintic efficacy of Cymbopogon citratus essential oil (CcEO) and C. citratus essential oil nanoemulsion (CcEOn). Pysicochemical analyses were performed. The in vitro effect was determined using the egg hatch test (EHT) on Haemonchus contortus and in vivo effect was evaluated in sheep infected with gastrointestinal nematodes. The animals were treated with CcEO (500 mg/kg) or CcEOn (450 mg/kg) for the fecal egg count (FEC) and the determination of worm burden. The main component of CcEO was citral. The CcEO content in the nanoemulsion was 20% (v/v), and the mean particle size was 248 nm. In EHT, CcEO and CcEOn (1.25 mg/mL) inhibited larval hatching by 98.4 and 97.1%, respectively. Three animals treated with CcEO died whereas in the group treated with CcEOn one animal died. The FEC and total worm burden of the treated groups did not differ from the negative control (p>0.05). The CcEOn showed efficacy only on H. contortus (p 0.05). In conclusion, nanoencapsulation reduced toxicity and increased efficacy on H. contortus.(AU)
A resistência anti-helmíntica estimulou a busca por estratégias de controle de nematoides gastrintestinais, incluindo óleos essenciais livres ou em nanoemulsão. Este estudo avaliou a eficácia anti-helmíntica do óleo essencial de Cymbopogon citratus (OECc) e da nanoemulsão do óleo essencial de C. citratus (nOECc). Análises físico-químicas foram realizadas. O efeito in vitro foi determinado no teste de eclosão de ovos (TEO) sobre Haemonchus contortus e o efeito in vivo foi avaliado em ovinos infectados com nematoides gastrintestinais. Os animais foram tratados com OECc (500 mg/kg) ou nOECc (450 mg/kg) para determinação do número de ovos por grama de fezes (OPG) e carga parasitária. O principal constituinte do OECc foi citral. O conteúdo de OECc na nanoemulsão foi 20% e o tamanho médio de partícula foi 248 nm. No TEO, OECc e nOECc (1,25 mg/mL) inibiram 98,4 e 97,1% da eclosão larvar, respectivamente. Três animais tratados com o OECc morreram, enquanto um animal do grupo tratado com a nOECc morreu. O OPG e a carga parasitária total dos grupos tratados não diferiram do controle negativo (p>0.05). A nOECc apresentou eficácia somente sobre H. contortus (p 0.05). Como conclusão, o nanoencapsulamento reduziu a toxicidade e aumentou a eficácia sobre H. contortus.(AU)
Assuntos
Animais , Óleos Voláteis/efeitos adversos , Óleos Voláteis/análise , Anti-Helmínticos/efeitos adversos , Anti-Helmínticos/análise , Cymbopogon , Nematoides , OvinosRESUMO
This study aimed to evaluate the anthelmintic activity of Eucalyptus citriodora essential oil and citronellal on sheep gastrointestinal nematodes. Essential oil composition was determined by gas chromatography mass spectrometry. The substances were evaluated in vitro using adult worm motility test (AWMT) and transmission electron microscopy (TEM). The acute toxicity test in mice and the fecal egg count reduction test (FECRT) in sheep were performed. Citronellal was confirmed as the essential oil major constituent (63.9%). According to the AWMT, 2 mg/mL of essential oil and citronellal completely inhibited Haemonchus contortus motility at 6 h post exposure. H. contortus exposed to essential oil and citronellal exhibited internal ultrastructural modifications. The lethal dose 50 values in mice were 5,000 and 2,609 mg/kg for essential oil and citronellal, respectively. E. citriodora essential oil reduced sheep epg at 14 days post treatment by 69.5% (P 0.05). No significant differences were observed in epg between the citronellal and negative control groups (P>0.05). The interaction between citronellal and other constituents in the essential oil may be relevant for its in vivo anthelmintic activity. Thus, E. citriodora essential oil and citronellal pharmacokinetic studies may help elucidate the anthelmintic activity of these compounds.(AU)
Este trabalho objetivou avaliar a atividade anti-helmíntica do óleo essencial de Eucalyptus citriodora e citronelal sobre nematoides gastrintestinais de ovinos. A composição do óleo essencial foi determinada por cromatografia gasosa acoplada à espectrometria de massas. As substâncias foram avaliadas in vitro utilizando-se teste de motilidade de vermes adultos (AWMT) e microscopia eletrônica de transmissão (TEM). Teste de toxicidade aguda em camundongos e teste de redução da contagem de ovos fecais (FECRT) em ovinos foram realizados. Citronelal foi confirmado como componente majoritário do óleo essencial (63,9%). No AWMT, 2 mg/mL de óleo essencial e citronelal inibiram completamente a motilidade de H. contortus 6 h pós-exposição. H. contortus expostos ao óleo essencial e citronelal exibiram modificações ultraestruturais internas. Os valores da dose letal 50 em camundongos foram 5.000 e 2.609 mg/kg para óleo essencial e citronelal, respectivamente. Óleo essencial de E. citriodora reduziu opg de ovinos 14 dias pós-tratamento em 69,5% (P 0,05). Não houve diferença significativa de opg entre grupo controle negativo e citronelal (P>0,05). A interação entre citronelal e outros constituintes do óleo essencial pode ser relevante na atividade anti-helmíntica in vivo. Portanto, avaliação farmacocinética do óleo essencial de E. citriodora e citronelal pode auxiliar a elucidar a atividade anti-helmíntica desses compostos.(AU)
Assuntos
Animais , Ovinos/parasitologia , Anti-Helmínticos/administração & dosagem , Anti-Helmínticos/análise , Gastroenteropatias/parasitologia , Óleos Voláteis , Eucalyptus/parasitologiaRESUMO
Background: Gastrointestinal nematodes are one of the major health and economic problem of sheep and goats in the world. The control of these nematodes is carried out conventionally with synthetic anthelminths, which favored the selection of gastrointestinal nematode (GIN) populations multiresistant to anthelmintics. The emergence of anthelmintic resistance has stimulated the search for new alternatives to control small ruminant GIN, standing out the use of plants and their bioactives compounds, such as essential oils (EO). The objective of this review was to present the main characteristics and anthelmintic activity of EO, their isolated compounds and drug delivery systems in the control of GIN.Review: Essential oils are a complex blend of bioactive compounds with volatile, lipophilic, usually odoriferous and liquid substances. EO are composed of terpenes, terpenoids, aromatic and aliphatic constituents. EO has various pharmacological activities of interest in preventive veterinary medicine such as antibacterials, antifungals, anticoccicids, insecticides and anthelmintics. In vitro and in vivo tests are used to validate the anthelmintic activity of EO on GIN. In vitro tests are low cost screening tests that allow the evaluation of the anthelmintic activity of a large amount of bioactive compounds on eggs, first (L1) and third stage larvae (L3), and adult nematodes. The antiparasitic effect of EO is related to its main compound or to the interaction of the compounds. These bioactive compounds penetrate the cuticle of the nematodes by transcuticular diffusion, altering the mechanisms of locomotion, besides causing cuticular lesions. Following in vitro evaluation, the acute and sub-chronic toxicity test should be performed to assess the toxicity of the bioactive compounds and to define the dose to be used in in vivo tests.[...]