Antihelminthic action of the Anethum graveolens essential oil on Haemonchus contortus eggs and larvae / Ação anti-helmítica do óleo essencial de Anethum graveolens sobre ovos e larvas de Haemonchus contortus

Abstract Gastrointestinal nematodes are responsible for great economic losses in sheep raising, and their control has long been carried out almost exclusively by the administration of anthelmintics, which have led to serious resistance problems. In the search for alternative control measures, phytotherapic research is highlighted. The aim of this study was to evaluate the action of Anethum graveolens (dill) essential oil on different stages of Haemonchus contortus life cycle, as well its cytotoxicity MDBK (Madin-Darby bovine kidney) cells. H. contortus larvae and eggs were obtained from infected sheep feces, and essential oil extracted from plant seeds through the Clevenger apparatus. 9.4, 4.7, 2.35, 1.17. 0.58 and 0.29 mg/mL concentrations were evaluated. The Egg Hatch Inhibition (HI), Larval Development Inhibition (LDI) and Larval Migration Inhibition (LMI) techniques were used. Thybendazole 0.025 mg/mL in HI and Levamisole 0.02 mg/mL in the LDI and LMI tests were used as positive controls, while distilled water and a Tween 80 solution were used as positive negative controls. The inhibition results obtained for the highest oil concentration were: HI 100%, LDI 98.58% and LMI 63.7%, differing (�� <0.05) from negative controls. Main A. graveolens oil components present in 95.93% of the total oil were Dihydrocarvone (39.1%), Carvone (22.24%), D-Limonene (16.84%), Apiol (10.49%) and Trans-dihydrocarvone (7.26%). Minimum A. graveolens essential oil concentrations required to inhibit 50% (IC50) of egg hatching, larval development and larval migration were 0.006 mg/mL, 2.536 mg/mL and 3.963 mg/mL, respectively. Cell viability in MDBK (Madin-Darby bovine kidney) cells, when incubated with A. graveolens essential oil, was 86% for the highest (9.4 mg/mL) and 99% for the lowest concentration (0.29 mg/mL). A. graveolens essential oil, according to the results obtained in this study, is a promising alternative in sheep gastrointestinal nematode control.

Resumo Os nematoides gastrintestinais são responsáveis ​​por grandes perdas econômicas na ovinocultura, e seu controle tem sido realizado quase exclusivamente pela administração de anti-helmínticos, que levaram a sérios problemas de resistência. Na busca de medidas alternativas de controle, destaca-se a pesquisa fitoterápica. O objetivo deste trabalho foi avaliar a ação do óleo essencial de Anethum graveolens (endro) em diferentes estágios de Haemonchus contortus, bem como testar a viabilidade celular para o óleo. Larvas e ovos de H. contortus foram obtidos de fezes de ovinos infectados e óleo essencial extraído de sementes de plantas através do aparelho de Clevenger. As concentrações avaliadas foram 9,4, 4,7, 2,35, 1,17, 0,58 e 0,29 mg/mL. Verificou-se a Inibição de eclosão dos ovos (IE), Inibição de Desenvolvimento Larval (IDL) e Inibição de Migração Larval (IML). Tiabendazol 0,025 mg/mL em IE e Levamisole 0.02 mg/mL nos testes IDL e IML foram usados ​​como controles positivos, enquanto água destilada e uma solução Tween 80 foram usados como controles negativos. Os resultados de inibição obtidos para a maior concentração de óleo foram: IE 100%, IDL 98,58% e IML 63,7%, diferindo (�� <0,05) dos controles negativos. Os principais componentes presentes em 95,93% do óleo total de A. graveolens foram Di-hidrocarvona (39,1%), Carvona (22,24%), D-Limoneno (16,84%), Apiol (10,49%) e Trans-di-hidrocarvona (7,26%). As concentrações mínimas de óleo essencial de A. graveolens necessárias para inibir 50% (IC50) de eclosão dos ovos, desenvolvimento larval e migração larval foram de 0,006 mg/mL, 2,536 mg/mL e 3,963 mg/mL, respectivamente. A viabilidade celular nas células MDBK (rim bovino Madin-Darby), quando incubadas com o óleo essencial de A. graveolens, foi de 86% para a maior (9,4 mg/mL) e 99% para a menor concentração (0,29 mg/mL). O óleo essencial de A. graveolens mostrou ser uma alternativa promissora no controle de nematoides gastrintestinais de ovinos.

Antihelminthic action of the Anethum graveolens essential oil on Haemonchus contortus eggs and larvae.

Gastrointestinal nematodes are responsible for great economic losses in sheep raising, and their control has long been carried out almost exclusively by the administration of anthelmintics, which have led to serious resistance problems. In the search for alternative control measures, phytotherapic research is highlighted. The aim of this study was to evaluate the action of Anethum graveolens (dill) essential oil on different stages of Haemonchus contortus life cycle, as well its cytotoxicity MDBK (Madin-Darby bovine kidney) cells. H. contortus larvae and eggs were obtained from infected sheep feces, and essential oil extracted from plant seeds through the Clevenger apparatus. 9.4, 4.7, 2.35, 1.17. 0.58 and 0.29 mg/mL concentrations were evaluated. The Egg Hatch Inhibition (HI), Larval Development Inhibition (LDI) and Larval Migration Inhibition (LMI) techniques were used. Thybendazole 0.025 mg/mL in HI and Levamisole 0.02 mg/mL in the LDI and LMI tests were used as positive controls, while distilled water and a Tween 80 solution were used as positive negative controls. The inhibition results obtained for the highest oil concentration were: HI 100%, LDI 98.58% and LMI 63.7%, differing (𝑝 <0.05) from negative controls. Main A. graveolens oil components present in 95.93% of the total oil were Dihydrocarvone (39.1%), Carvone (22.24%), D-Limonene (16.84%), Apiol (10.49%) and Trans-dihydrocarvone (7.26%). Minimum A. graveolens essential oil concentrations required to inhibit 50% (IC50) of egg hatching, larval development and larval migration were 0.006 mg/mL, 2.536 mg/mL and 3.963 mg/mL, respectively. Cell viability in MDBK (Madin-Darby bovine kidney) cells, when incubated with A. graveolens essential oil, was 86% for the highest (9.4 mg/mL) and 99% for the lowest concentration (0.29 mg/mL). A. graveolens essential oil, according to the results obtained in this study, is a promising alternative in sheep gastrointestinal nematode control.

Saccharomyces cerevisiae (YT001) supplementation for the control of Haemonchus contortus and modulation of the immune response of sheep.

Studies aiming at the development and evaluation of alternative methods to minimise losses caused by the gastrointestinal nematode Haemonchus contortus are extremely important. Such research is essential, given the high morbidity rates among sheep and the significant mortality rates of lambs, allied to the low efficacy of commercial products for the control of this parasite. The purpose of this study was to evaluate the effect of the Saccharomyces cerevisiae (YT001 - YEASTECH) on the control of H. contortus and its modulation of the immune response in experimentally infected sheep. Eighteen sheep were divided into two groups. Group 1, the control group, comprised animals infected with H. contortus and supplemented with distilled water, while Group 2, the treated group, consisted of animals infected and supplemented with S. cerevisiae (400 million cfu/day of suspension for 49 days). The following parasitological parameters were evaluated: number of eggs per gram of faeces, number of infective larvae (L3) recovered per faecal culture, and parasitic load of the abomasum. The following immunological parameters were quantified: immunoglobulin (Ig)A in the mucous secretions and serum IgG; cytokines interleukin (IL)-4, IL-5 and IL-10; number of eosinophils in the abomasal mucosa and groups of cells positive for the markers: MHCII, CD4+CD25+, CD5+CD8+, WC4, CD5+CD4+, CD8+CD11b+ and CD5+WC1 by whole blood flow cytometry. The results revealed a significant decrease (P<0.05) in the number of larvae and significantly higher serum IgG levels (P<0.05) in the group supplemented with S. cerevisiae. The supplemented animals showed significantly larger numbers of eosinophils (P<0.05), as well as more cells positive for MHCII, CD4+CD25+, CD5+CD8+ than the control animals. This study confirmed the beneficial action of S. cerevisiae on the host immune response to H. contortus, as evidenced mainly by the smaller number of L3 recovered from the faeces of sheep supplemented with S. cerevisiae.

Antihelminthic action of the Anethum graveolens essential oil on Haemonchus contortus eggs and larvae

Abstract Gastrointestinal nematodes are responsible for great economic losses in sheep raising, and their control has long been carried out almost exclusively by the administration of anthelmintics, which have led to serious resistance problems. In the search for alternative control measures, phytotherapic research is highlighted. The aim of this study was to evaluate the action of Anethum graveolens (dill) essential oil on different stages of Haemonchus contortus life cycle, as well its cytotoxicity MDBK (Madin-Darby bovine kidney) cells. H. contortus larvae and eggs were obtained from infected sheep feces, and essential oil extracted from plant seeds through the Clevenger apparatus. 9.4, 4.7, 2.35, 1.17. 0.58 and 0.29 mg/mL concentrations were evaluated. The Egg Hatch Inhibition (HI), Larval Development Inhibition (LDI) and Larval Migration Inhibition (LMI) techniques were used. Thybendazole 0.025 mg/mL in HI and Levamisole 0.02 mg/mL in the LDI and LMI tests were used as positive controls, while distilled water and a Tween 80 solution were used as positive negative controls. The inhibition results obtained for the highest oil concentration were: HI 100%, LDI 98.58% and LMI 63.7%, differing ( 0.05) from negative controls. Main A. graveolens oil components present in 95.93% of the total oil were Dihydrocarvone (39.1%), Carvone (22.24%), D-Limonene (16.84%), Apiol (10.49%) and Trans-dihydrocarvone (7.26%). Minimum A. graveolens essential oil concentrations required to inhibit 50% (IC50) of egg hatching, larval development and larval migration were 0.006 mg/mL, 2.536 mg/mL and 3.963 mg/mL, respectively. Cell viability in MDBK (Madin-Darby bovine kidney) cells, when incubated with A. graveolens essential oil, was 86% for the highest (9.4 mg/mL) and 99% for the lowest concentration (0.29 mg/mL). A. graveolens essential oil, according to the results obtained in this study, is a promising alternative in sheep gastrointestinal nematode control.

Resumo Os nematoides gastrintestinais são responsáveis por grandes perdas econômicas na ovinocultura, e seu controle tem sido realizado quase exclusivamente pela administração de anti-helmínticos, que levaram a sérios problemas de resistência. Na busca de medidas alternativas de controle, destaca-se a pesquisa fitoterápica. O objetivo deste trabalho foi avaliar a ação do óleo essencial de Anethum graveolens (endro) em diferentes estágios de Haemonchus contortus, bem como testar a viabilidade celular para o óleo. Larvas e ovos de H. contortus foram obtidos de fezes de ovinos infectados e óleo essencial extraído de sementes de plantas através do aparelho de Clevenger. As concentrações avaliadas foram 9,4, 4,7, 2,35, 1,17, 0,58 e 0,29 mg/mL. Verificou-se a Inibição de eclosão dos ovos (IE), Inibição de Desenvolvimento Larval (IDL) e Inibição de Migração Larval (IML). Tiabendazol 0,025 mg/mL em IE e Levamisole 0.02 mg/mL nos testes IDL e IML foram usados como controles positivos, enquanto água destilada e uma solução Tween 80 foram usados como controles negativos. Os resultados de inibição obtidos para a maior concentração de óleo foram: IE 100%, IDL 98,58% e IML 63,7%, diferindo ( 0,05) dos controles negativos. Os principais componentes presentes em 95,93% do óleo total de A. graveolens foram Di-hidrocarvona (39,1%), Carvona (22,24%), D-Limoneno (16,84%), Apiol (10,49%) e Trans-di-hidrocarvona (7,26%). As concentrações mínimas de óleo essencial de A. graveolens necessárias para inibir 50% (IC50) de eclosão dos ovos, desenvolvimento larval e migração larval foram de 0,006 mg/mL, 2,536 mg/mL e 3,963 mg/mL, respectivamente. A viabilidade celular nas células MDBK (rim bovino Madin-Darby), quando incubadas com o óleo essencial de A. graveolens, foi de 86% para a maior (9,4 mg/mL) e 99% para a menor concentração (0,29 mg/mL). O óleo essencial de A. graveolens mostrou ser uma alternativa promissora no controle de nematoides gastrintestinais de ovinos.

Bacillus spp. toxicity against Haemonchus contortus larvae in sheep fecal cultures.

The gastrointestinal nematode Haemonchus contortus is a major productivity constraint in sheep. In this study, the nematicidal effects of Bacillus circulans, Bacillus cereus, Bacillus thuringiensis var. israelensis, Bt. var. osvaldocruzi, Bt. var. morrisoni, and Bt. var. kurstaki were assessed in free-living larval stages of H. contortus. A spore-crystal suspension containing approximately 2×10(8)UFC/mL of each strain was added to sheep feces that were naturally infected with H. contortus eggs, and the presence of larvae was then evaluated. We observed a significant (p>0.05) reduction in larval development when using B. circulans, B. thuringiensis var. israelensis, Bt. var. osvaldocruzi and Bt. var. kurstaki, and these effects were proportional with the amount of bacteria added to the feces. However, no effect was observed when Bt. var. morrisoni or B. cereus was added. These observations suggest that these bacteria might be effective as nematicides and may allow for the development of integrated biological control of zooparasitic nematodes.