Resumen Haemonchus contortus es un nematodo hematófago con alta tasa de reproducción, considerado como el principal problema en pequeños rumiantes en pastoreo. Por tanto, se buscan alternativas de tratamiento con base en el uso de los extractos vegetales. El objetivo de este estudio fue evaluar la actividad ovicida de Artemisia cina contra el parásito Haemonchus contortus y caracterizar químicamente el extracto con mayor actividad biológica a través de cromatografía de gases acoplado a espectrometría de masas (CG-MS). La obtención de los extractos a evaluar, se realizó con la técnica de maceración utilizando metanol, acetato de etilo y n-hexano. Los extractos se llevaron a sequedad total y se desafiaron frente a los huevos de H. contortus utilizando la técnica de inhibición de la eclosión de huevos descrita por la Asociación Mundial para el Avance de la Parasitología Veterinaria (WAAVP, por sus siglas en inglés). El extracto metanólico (EM) mostró el 100 % de actividad ovicida a una concentración de 4.25 mg/ml, siendo el más activo a una baja concentración, por tanto se caracterizó con CG-MS. EL EM posee principalmente ácidos grasos y terpenos; de ellos, el ácido hexadecanoico y el 2-[4-metil-6-(2,6,6-trimetilciclohex-1-enil) exa-1,3,5-trienil][ciclohex]-1-en-carboxialdehido. Los compuestos caracterizados han mostrado actividad antihelmíntica previamente reportada, por lo que la actividad ovicida puede estar asociada a estos. En conclusión, el extracto metanólico de A. cina tuvo una mayor actividad ovicida a bajas concentraciones, esto se debe probablemente a la presencia de ácidos grasos y terpenos.

Abstract Haemonchus contortus is a hematophagous nematode with a high reproduction rate, considered to be the main problem in grazing small ruminants. Therefore, treatment alternatives based on the use of plant extracts are sought. This study aimed to evaluate the ovicidal activity of Artemisia cina against the parasite Haemonchus contortus and to chemically characterize the extract with the highest biological activity through gas chromatography coupled with mass spectrometry (GC-MS). The extracts to be evaluated were obtained through the maceration technique using methanol, ethyl acetate, and n-hexane. The extracts were taken to total dryness and challenged against H. contortus eggs using the egg-hatching inhibition technique described by the World Association for the Advancement of Veterinary Parasitology (WAAVP). The methanolic extract (ME) showed 100 % ovicidal activity at a concentration of 4.25 mg/ml, being the most active at a low concentration; therefore, it was characterized by GC-MS. ME mainly contains fatty acids and terpenes; among them are hexadecanoic acid and 2-[4-methyl-6-(2,6,6-trimethylcyclohex-1-enyl)hexa-1,3,5-trienyl][cyclohex]-1-en-carboxyaldehyde. The characterized compounds have shown previously reported anthelmintic activity so that ovicidal activity may be associated with them. In conclusion, the methanolic extract of A. cina had a higher ovicidal activity at low concentrations; this is probably due to the presence of fatty acids and terpenes.

Abstract The aim of this study was to assess the in vitro ovicidal activity of an ethyl acetate extract from Arceuthobium vaginatum (EtOAc-E) and their subfractions (AvR5-AvR14) against Haemonchus contortus using the egg hatching inhibition (EHI) test. The EtOAc-E and subfractions were tested at 0.12-2.00 and at 0.015-2.0 mg/mL, respectively. Distilled water and methanol (2%) were used as negative controls and Thiabendazole (0.10 mg/mL) as a positive control. Treatments with a dependent effect on concentration were subjected to regression analysis to determine the effective concentrations (EC50 and EC90). The major secondary compounds present in the extract and subfractions were identified by high performance liquid chromatography (HPLC). The EtOAc-E and AvR9 exhibited the best ovicidal effect recording 97.5 and 100% of EHI at 0.25 mg/mL, respectively. The EtOAc-E and AvR9 displayed an EC50= 0.12 and 0.08 mg/mL, respectively. The HPLC analysis in the EtOAc-E and bioactive fractions indicated the presence of a polyphenol, glycosylated flavanones, quercetin glucoside, cinnamates, coumarin, cinnamic acid derivative, ferulic acid, coumarate, naringenin, protocatechuic acid and naringin. Results demonstrated that A. vaginatum extract and fraction is able to inhibit the egg hatch process of H. contortus and could be a viable option for the control of small ruminant haemonchosis.

Resumo O objetivo deste estudo foi avaliar a atividade ovicida in vitro de um extrato de acetato de etila de Arceuthobium vaginatum (EtOAc-E) e suas subfrações (AvR5-AvR14) contra Haemonchus contortus, utilizando-se o teste de inibição da eclosão de ovos (EHI). O extrato foi testado em 0.12-2.00 mg/mL, e as subfrações foram testadas em 0.15-2.0 mg/mL. Água destilada e metanol (2%) foram utilizados como controles negativos e Tiabendazol (0.10 mg/mL) como controle positivo. Os tratamentos com efeito concentração dependente foram submetidos à análise de regressão para determinação das concentrações efetivas (CE50 e CE90). Os principais compostos secundários, presentes no extrato e nas subfrações, foram identificados por cromatografia líquida de alta eficiência (HPLC). EtOAc-E e subfração AvR9 exibiram o melhor efeito ovicida, registrando-se 97.5 e 100% do EHI a 0.25 mg/mL, respectivamente. O EtOAc-E e AvR9 apresentaram CE50= 0,12 e 0,08 mg/mL, respectivamente. A análise por HPLC nas frações EtOAc-E e bioativas indicou a presença de um polifenol, flavanonas glicosiladas, glicosídeo de quercetina, cinamatos, cumarina, derivado de ácido cinâmico, ácido ferúlico, cumarato, naringenina, ácido protocatecuico e naringina. Os resultados demonstraram que extrato e frações de A. vaginatum são capazes de inibir o processo de eclosão dos ovos de H. contortus, podendo ser uma opção viável para o controle da hemoncose em pequenos ruminantes.

Abstract This study aimed to evaluate the effect of supplementing the maintenance diet of Boer goat kids with Acacia cochliacantha leaves. The endpoints evaluated were Haemonchus contortus fecal egg count (FEC) and water and dry matter intake. Two experimental treatments were evaluated on ten recently weaned goat kids (16.850 ± 1.630 kg of initial live weight and three months of age) experimentally infested with H. contortus larvae (L3) (350 larvae per live weight kilogram). Treatment 1 (T1) served as the control and consisted of infested animals without diet supplementation with A. cochliacantha leaves. Treatment 2 (T2) consisted of infested animals fed diets supplemented with 5% of A. cochliacantha leaves. Animals were grouped from highest to lowest based on their FEC. The two animals groups with the highest values were randomly assigned to T1 or T2; this was repeated until completing five repetitions per treatment. The evaluated variables were: FEC (per gram of feces), water intake, and dry matter intake (DMI). The results show that goat kids fed diets with 5% of A. cochliacantha leaves have lower (P<0.05) FEC than the control. There were no significant differences in water intake and DMI (g d-1) between treatments. This study demonstrates the anthelmintic activity of diets supplemented with A. cochliacantha leaves in goat kids. Thus, this arboreal legume could represent a viable option for the comprehensive management of the nematodiasis of growing Boer goat kids.

Resumen: El objetivo del presente trabajo fue evaluar el efecto de la inclusión de hojas de Acacia cochliacantha en una dieta de mantenimiento para cabritos de la raza Boer en el conteo fecal de huevos (CFH) de Haemonchus contortus, consumo de agua y materia seca. Diez cabritos recién destetados (16.850 ± 1.630 kg de peso vivo inicial y 3 meses de edad), fueron experimentalmente infestados con larvas (L3) de H. contortus (350 larvas por kilogramo de peso vivo), para probar dos tratamientos: T1: testigo (animales infectados con larvas L3 de H. contortus, sin inclusión de hojas de A. cochliacantha) y T2: animales infectados con larvas L3 de H. contortus, con inclusión de 5% hojas de A. cochliacantha en la dieta. Antes de asignar los tratamientos a los animales, se realizó un CFH por gramos de heces en cada uno de ellos y de esta manera agruparlos de mayor a menor en relación a la cantidad de huevos presentes en las heces, en los dos animales con valores más altos se asignaron aleatoriamente T1 o T2 y así sucesivamente hasta completar cinco repeticiones verdaderas para cada tratamiento. Las variables medidas fueron: CFH (por gramo de heces), consumo de agua y consumo de materia seca (CMS). Los resultados encontrados demostraron reducción (P<0.05) en el CFH de H. contortus en los cabritos donde se incluyó el 5% de hojas de A. cochliacantha en su dieta. Para el consumo de agua y CMS (g día-1) no existieron diferencias entre tratamientos (P> 0.05). Se concluye que la adición de hojas de A. cochliacantha en dietas para cabritos tienen actividad antihelmíntica, por lo que esta leguminosa arbórea podría representar una opción en el manejo integral de la nematodiasis de cabritos Boer en crecimiento.

Abstract The ovicidal effect of hydro-alcoholic extracts from mature fruits and dried leaves of the Caesalpinia coriaria tree, against Haemonchus contortus and H. placei was evaluated by the egg hatching inhibition test (EHIT). Secondary compounds within the extracts were identified by high performance liquid chromatography. Five extract concentrations were assessed in the inhibition tests (25.0, 12.5, 6.2, 3.1 and 1.5 mg/mL), using distilled water and Thiabendazole (0.5 mg/mL) as negative and positive controls, respectively. The results were analysed under a 2 × 2 × 6 completely randomized factorial design, and mean differences were established by the Tukey’s post hoc test. The 50% (EC50) and 90% (EC90) effective concentrations for each extract were determined using a Probit analysis. A concentration-dependent ovicidal effect of hydro-alcoholic extracts from both leaves and fruits was observed, reaching a 100% efficacy against both nematode species with the 25.0 mg/mL concentration (p < 0.05). The EC50 for the fruit and leaves extracts were established as 1.63 and 3.91 mg/mL, and as 3.98 and 11.68 mg/mL, against H. contortus and H. placei, respectively. The most important secondary compounds identified in extracts were free-condensed tannins, gallic acid and methylgallate. In conclusion, hydro-alcoholic extracts from C. coriaria mature fruits and dried leaves inhibit in vitro egg hatching of H. contortus and H. placei.

ABSTRACT Moussonia deppeana (Schltdl. & Cham.) Klotzsch ex Hanst., Gesneriaceae, known as tlachichinole, is a Mexican medicinal plant used for treatment of chronic inflammation-related diseases such as arthritis. In this paper, the main metabolite verbascoside was quantified in ethanolic extract; anti-arthritic and antioxidant activities were also evaluated in Complete Freund's Adjuvant induced arthritis in mice, with complete hematological evaluation, and oxidative stress measure in edema and ganglionic tissues on day 28. In popliteal ganglion, CD4+ lymphocytes and tumor necrosis factor alpha concentration were measured in addition to histological analysis. Ethanolic extract contained 79.2 mg of verbascoside/g extract, and this extract at 450 mg/kg generated an inhibition of 24% over paw edema development and increased body weight gain on final day. For hematological parameters, same dose decreased total leukocytes and lymphocytes, as well as decreased oxidation rate over biomolecules in edema and ganglionic tissues, and increased antioxidant enzyme activity. In ganglionic tissue, CD4+ lymphocytes and tumor necrosis factor alpha level showed no differences at any tested dose compared to complete Freund's adjuvant untreated group. Histological analysis of popliteal ganglion revealed moderate reduction of follicular hyperplasia, leukocyte infiltration and lipid inclusions at 450 mg/kg dose. Ethanolic extract of M. deppeana possesses anti-edematous activity associated to a moderate reduction in follicular hyperplasia, with immune-modulatory and antioxidant effects during experimental arthritis in mice.

Abstract Bioactive plants might represent an alternative for Cooperia punctata control in grazing cattle. The objectives of this study were (1) to assess the ovicidal activity of extracts from 4 plant species against C. punctata, (2) to determine the role of the polyphenols in the plants’ anthelmintic (AH) activity, and (3) to evaluate the best plant extraction procedure when searching for ovicidal activity. The egg hatch assay was used with different extraction procedures, aqueous (AQ), acetone:water (AW) and acetonic (AC), to evaluate the ovicidal activity of Leucaena leucocephala, Gliricidia sepium, Guazuma ulmifolia and Cratylia argentea. Eggs of C. punctata were exposed in quadruplicate to 0.6, 1.2, 2.4, 4.8 and 9.6 mg mL-1 of each plant extract. The roles of the polyphenols were assessed using polyethylene glycol (PEG). The 12 plant extracts inhibited egg hatching in a dose-dependent manner. Best-fit LC50 values were 1.03 ± 0.17 and 7.90 ± 1.19 mg mL-1 for G. sepium-AC and L. leucocephala-AQ, respectively. Differences in AH activity were found among the extraction procedures (P < 0.05). At the highest concentration, L. leucocephala-AQ inhibited more than 50% of C. punctata hatching. The G. sepium-AC extract fully inhibited egg hatching. The addition of polyethylene glycol revealed the role of the polyphenols in the bioactivity of most plant extracts; however, for G. sepium-AC, the polyphenols were not the main bioactive compounds. Overall, acetone:water extraction represented the best extraction procedure to obtain both ovicidal activity and higher yield. The inhibition rates suggested that L. leucocephala and G. sepium should be evaluated as a means of reducing larval density in pastures.

Resumen El uso de plantas bioactivas representa una alternativa para el control de Cooperia punctata en el ganado de pastoreo. Los objetivos de este estudio fueron: (1) evaluar la actividad ovicida de extractos de cuatro especies de plantas contra C. punctata, (2) determinar la participación de los compuestos polifenólicos (CP) en la actividad antihelmíntica (AH) y (3) evaluar el mejor método de extracción para alcanzar una actividad ovicida. El bioensayo de inhibición de la eclosión de huevos (IEH) fue utilizado para evaluar el efecto ovicida de Leucaena leucocephala, Gliricidia sepium, Guazuma ulmifolia y Cratylia argentea; con diferentes métodos de extracción: acuoso (AQ), acetona:agua (AW) y acetónico (AC). Los huevos de C. punctata fueron incubados por cuadruplicado a 0.6, 1.2, 2.4, 4.8 y 9.6 mg mL-1 de cada uno de los extractos de plantas. La participación de los CP fue determinada mediante el uso de polietilenglicol (PEG). Los 12 extractos de plantas inhibieron la eclosión de huevos de modo dosis-dependiente, y las mejores concentraciones letales medias (CL50) fueron: 1.03 ± 0.17 para G. sepium-AC, y 7.90 ± 1.19 mg mL-1 para L. leucocephala-AQ. Se observaron diferencias en la actividad AH según el método de extracción (P < 0.05). A mayor concentración, L. leucocephala-AQ inhibió más del 50 % de la eclosión de C. punctata, entretanto, el extracto G. sepium-AC inhibió la eclosión en su totalidad. La adición de PEG demostró la participación de los CP en la actividad AH de la mayoría de los extractos; no obstante, con G. sepium-AC, los CP no fueron los principales compuestos bioactivos. En general, el mejor método de extracción fue acetona:agua, pues permitió obtener una buena actividad ovicida, con un mayor rendimiento del extracto. Las tasas de inhibición sugieren que L. leucocephala y G. sepium podrían ser evaluadas con la finalidad de reducir la densidad larvaria en pastos.

Two new triterpene saponins whose aglycons are based on the oleanane skeleton (1-2), were isolated from the roots of Viguiera hypargyrea, together with two known triterpene saponins (3 and 4) as well as the triterpenes friedelin, friedelan 3β-ol and oleanolic acid. The structures of the new compounds were established mainly by 2D NMR techniques of their peracetylated derivatives as 3-O-[α-L-rhamnopyranosyl (1 → 3)-β-D-xylopyranosyl (1 → 4)]-β-D-glucopyranosyl-oleanolic acid-28-O-β-D-glucopyranoside and 3-O-[α-L-rhamnopyranosyl (1 → 3)-β-D-xylopyranosyl (1 → 4)]-β-D-glucopyranosyl oleanolic acid respectively.

De las raíces de Viguiera hypargyrea se aislaron dos nuevas saponinas triterpénicas (1-2), cuyas agliconas corresponden al esqueleto del oleanano, junto con dos saponinas triterpénicas conocidas (3 y 4), así como los triterpenos friedelina, friedelan-3β-ol y ácido oleanólico. Las estructuras de los compuestos novedosos fueron establecidas principalmente por medio de técnicas de RMN 2D de sus derivados peracetilados como 3-O-[α-L-rhamnopiranosil (1 → 3)-β-D-xilopiranosil (1 → 4)]-β-D-glucopiranosil-ácido oleanólico-28-O-β-D-glucopiranósido y ácido 3-O-[α-L-ramnopiranosil (1 → 3)-β-D-xilopiranosil (1 → 4)]-β-D-glucopiranosil oleanólico respectivamente.