Abstract Galanthamine is an Amaryllidaceae-derived acetylcholinesterase inhibitor used to treat memory impairment in Alzheimer's disease and vascular dementia. There is evidence that galanthamine, in addition to its effects on acetylcholinesterase, may enhance or inhibit brain nicotinic acetylcholine receptors, which could increase or decrease the therapeutic efficacy of galanthamine, respectively. Here, we evaluated the effects of galanthamine and two others Amaryllidaceae acetylcholinesterase inhibitors (haemanthamine and tazettine) analyzed by gas chromatography–mass spectrometry and identified by comparing their mass fragmentation patterns with literature and database NIST vs.2.0 on the agonist responses of brain nicotinic acetylcholine receptors α7, α3β4, (α4)2(β2)3 and (α4)3(β2)2. Using nicotinic acetylcholine receptors expressed heterologously in Xenopus oocytes, in conjunction with two-electrode voltage clamping, we found that galanthamine inhibits the function of nicotinic acetylcholine receptors assayed through a mix competitive and non-competitevely. Nicotinic acetylcholine receptor α7 were significantly more sensitive to inhibition (17 ± 0.6 µM) than the heteromeric receptor, α3β4 (90 ± 3.4 µM). Neither haemanthamine nor tazettine were more potent than galanthamine.

Abstract Flavonoid biosynthesis may be affected by plant domestication, with flavonoid production being reduced in proportion to the degree of domestication. In this context, kaempferol (3,4´,5,7-tetrahydroxyflavone) has been identified in the leaves of wild and cultivated Ugni molinae, a berry endemic to Chile. The biosynthetic pathway of kaempferol production begins with naringenin (4´,5,7-trihydroxyflavanone), which is converted to dihydrokaempferol (3,4´,5,7-tetrahydroxyflavanone), catalyzed by flavanone 3ß-hydroxylase (FHT) and then converted to kaempferol by a bifunctional enzyme called flavonol synthase (FLS). Therefore, our study aims to evaluate how FLS activity is affected in murtilla plants that are subjected to the domestication process. Kaempferol was quantified from methanolic extracts of leaf samples collected from both cultivated and wild U. molinae plants using high-performance liquid chromatography, and enzyme extraction was performed to determine FLS activity. The results showed that kaempferol concentration in wild plants from the Soloyo (0.14 μg g-1), Mehuín (0.18 μg g-1) and Queule (0.25 μg g-1) sampling areas was higher than in their cultivated counterparts. Our data are consistent with the FLS activity detected in samples obtained from Manzanal Alto (134.79 pKatal, Soloyo (96.48 pKatal), and Mehuín (119.97 pKatal). These samples also exhibited higher enzymatic activity than their cultivated counterparts. Together, these data suggest that FLS activity is negatively affected by the domestication process.

Resumen La biosíntesis de flavonoides puede verse afectada por la domesticación vegetal, reduciéndose la producción en relación al grado de domesticación. En este contexto, ha sido identificado el kaempferol (3,4´, 5,7-tetrahidroxiflavona) en hojas de Ugni molinae silvestre y cultivada. La ruta biosintética del kaempferol comienza con naringenina (4´, 5,7-trihidroxiflavanona), que se convierte en dihidrokaempferol (3,4´, 5,7-tetrahidroxiflavanona), catalizada por la flavanona ٣ß-hidroxilasa (FHT). Finalmente, se convierte en kaempferol por una enzima bifuncional llamada flavonol sintasa (FLS). Por lo tanto, nuestro estudio tiene como objetivo evaluar cómo se ve afectada la actividad de FLS en plantas de murtilla que están sujetas a un proceso de domesticación. El kaempferol fue cuantificado a partir de los extractos metanólicos de hojas recogidas en plantas cultivadas y silvestres de U. molinae mediante cromatografía líquida de alta resolución. Además, se realizó una extracción de enzimas para determinar la actividad de FLS. Los resultados mostraron que la concentración de kaempferol en las áreas de muestreo silvestre de Soloyo (0.14 μg g-1), Mehuín (0.18 μg g-1) y Queule (0.25 μg g-1) fue mayor que en sus contrapartes cultivadas. Estos datos fueron consistentes con la actividad de FLS determinada en muestras obtenidas de Manzanal Alto (134,79 pKatal), Soloyo (96,48 pKatal) y Mehuín (119,97 pKatal). Estos también exhibieron una mayor actividad enzimática que sus contrapartes cultivadas. Por lo tanto, nuestros datos sugieren que la actividad de FLS se ve reducida por la domesticación vegetal.

Abstract: In laboratory by dual choice test, stem and root alkaloid extracts of Berberis microphylla showed antifeedant activity against third-instar Plutella xylostella larvae. Stem extracts showed significantly lower leaf consumptions at concentrations of 500 and 1000 mg/L, as illustrated by antifeedant index of 72 and 97% respectively, whereas, root extracts were significantly active at 100, 500 and 1000 mg/L with antifeedant index of 68, 85 and 99% respectively. In addition, the pure berberine identified as the major alkaloid in the plant extracts showed significant lower food consumption by larvae only at concentration of 1000 mg/L, but the pure palmatine not affect feeding at any concentration. The results suggest that alkaloids extract of B. microphylla have promising antifeedant activity and could by incorporated in an integrated pest management (IPM) programs for P. xylostella.

Resumen: En laboratorio, mediante prueba de doble elección los extractos de alcaloides obtenidos de tallos y raíces de Berberis microphylla mostraron actividad antialimentaria frente a larvas de tercer estadio de Plutella xylostella. Los extractos de tallos mostraron consumos foliares significativamente más bajos en concentraciones de 500 y 1000 mg/L con un índice antialimentario de 72 y 97% respectivamente, mientras que los extractos de raíces fueron significativamente más activos a 100, 500 y 1000 mg/L con un índice antialimentario de 68, 85 y 99% respectivamente. Además, berberina identificada como el principal alcaloide de los extractos tuvo significativamente un menor consumo de alimento por parte de las larvas mostrando su efecto solo a la concentración de 1000 mg/L. Por otra parte, palmatina no tuvo efecto en la conducta de alimentación de las larvas a ninguna de las concentración evaluadas. Los resultados sugieren que los extractos de alcaloides obtenidos de B. microphylla tienen una prometedora actividad antialimentaria y podrían ser incorporados en un programa de manejo integrado de plagas (MIP) para P. xylostella.

ABSTRACT Acetylcholinesterase is an important target for control of neurodegenerative diseases causing cholinergic signaling deficit. Traditionally, galanthamine has been used as an Amaryllidaceae-derived acetylcholinesterase inhibitor, although new Amaryllidaceae plants could serve as source for better acetylcholinesterase inhibitors. Therefore, the objective of this study was to characterize the alkaloid composition from bulbs of Rhodolirium andicola (Poepp.) Traub, a native Chilean Amaryllidaceae specie, and assess their inhibitory activity on acetylcholinesterase by in vitro and in silico methodologies. Alkaloidal extracts from R. andicola exhibited an inhibitory activity with IC50 values between 11.25 ± 0.04 and 57.78 ± 1.92 µg/ml that included isolated alkaloid, galanthamine (2.3 ± 0.18 µg/ml), Additionally, 12 alkaloids were detected using gas chromatography–mass spectrometry and identified by comparing their mass fragmentation patterns with literature and database NIST vs.2.0. To better understand the bioactivity of isolated compounds and alkaloidal extracts against acetylcholinesterase, a molecular docking approach was performed. Results suggested that alkaloids such as lycoramine, norpluvine diacetate and 6α-deoxy-tazettine expand the list of potential acetylcholinesterase inhibitors to not only galanthamine. The role of R. andicola as a source for acetylcholinesterase inhibitors is further discussed in this study.

In the last decade, there has been increasing interest in investigating the impact of flavonoids on insects, specifically for pest control. In this study, we investigated the impact of isoflavonoids upon the feeding behavior of the clover root borer, Hylastinus obscurus Marsham (Coleoptera: Curculionidae), which is one of the most serious global pests associated with red clover, Trifolium pratense L. Four aglycones isoflavonoids: genistein (1), formononetin (2), daidzein (3) and biochanin A (4) were isolated and identified by HPLC, from roots of two Chilean red clover cultivars. The first two compounds, formononetin (2) and genistein (), showed high feeding deterrent activity when they were evaluated in artificial diets. This antifeedant effect of isoflavones on feeding behavior of H. obscurus suggests that they are responsible for a decreasedin insect weight gain as compared with the control. This information could be useful respectively, to farmers and researcher to produce and create plants resistant to curculionid.

ABSTRACT Previous researches have established that the swim speed of some microcrustaceans is influenced by chemical compounds emitted by conspecifics. We examined the hypothesis that cuticular compounds present on the body surface of A. franciscana, the most widespread member of Artemia genus, play a role in the swim speed of conspecific males. The movements (swim) of one male confronted to a sponge soaked with female or male cuticular extract, were recorded during 30 minutes and the swim speed was determined using a behavioral tracking software (Ethovision 3.1, Noldus Technologies). As a control, the movements of one male confronted to a sponge soaked with salty water or with a mixture of the solvents used in the extraction (chloroform-methanol), was recorded. The results showed that cuticular compounds from either female or male increase 1.5 (ca.) times the swim speed of males in comparison with the controls treatments salty water and the solvents. There was no a significant difference between the controls (salty water and chloroform-methanol). Chemical characterization was developed by sterification of the cuticular extracts and analyses by GC-FID and GC-MS. Four saturated fatty acid (myristic acid, palmitic acid, estearic acid, arachidic acid) and five insaturated fatty acids (oleic acid, linoleic acid, linoleic acid 3n3, cis-11-eicosanoic acid, euric acid) were identified. Myristic acid was found in female cuticular extract, but not in male cuticular extract. Results suggest that chemical compounds present in the cuticular surface of A. franciscana females could have an important role in the intra-specific recognition in this specie.

RESUMEN Investigaciones anteriores han establecido que la velocidad de nado de algunos microcrustáceos se encuentra influenciada por compuestos químicos emitidos por conespecíficos. Nosotros examinamos la hipótesis de que los compuestos cuticulares presentes sobre la superficie corporal de A. franciscana, el miembro con mayor distribución mundial del genero Artemia, juega un rol en la velocidad de nado de machos conespecíficos. Los movimientos nado de un macho confrontado a una esponja empapada con extracto cuticular de una hembra o de un macho, fueron grabados durante 30 minutos y la velocidad de nado fue determinada usando un software de rastreo conductual (Ethovision 3.1, Noldus Technologies). Como control, fueron grabados los movimientos de un macho confrontado a una esponja empapada con agua de mar artificial o con una mezcla de los solventes usados en la extracción (cloroformo-metanol). Los resultados mostraron que los compuestos cuticulares tanto de hembras como de machos incrementan 1,5 veces (aproximadamente) la velocidad de nado de los machos, en comparación con los experimentos control donde se uso agua de mar artificial y los solventes de la extracción. No hubo diferencias significativas entre los controles (agua de mar artificial y cloroformo-metanol). La caracterización química fue desarrollada mediante esterificación de los extractos cuticulares y analizados por CG-DIL y CG-EM. Cuatro ácidos grasos saturados (ácido mirístico, ácido palmítico, ácido esteárico, ácido araquídico) y cinco ácidos grasos insaturados (ácido oleico, ácido linoleico, ácido linoleico 3n3, ácido cis-11-eicosanoico, ácido eurico) fueron identificados. El ácido mirístico fue encontrado en los extractos cuticulares de hembras pero no en los extractos cuticulares de machos. Los resultados sugieren que los compuestos químicos presentes en la superficie cuticular de las hembras de A. franciscana podrían tener un importante rol en el reconocimiento intraespecífico de la especie.

The contents of long-chain fatty acids in non-polar root extracts from 1.6 year-old red clover plants of two cultivars (Superqueli INIA and Quiñequeli INIA) and one experimental line (Syn II Pre III), their relationships with H. obscurus populations, and the stimulant or deterrent effects of the root extracts and of individual fatty acids on the weight gain of H. obscurus were investigated. There was an inverse relationship between the insect population size and the palmitic acid content. The Superqueli INIA cultivar contained the highest palmitic acid content and the lowest insect load. When solutions of either palmitic or oleic acids were individually incorporated into an artificial diet, palmitic acid caused a decrease in weight gain, whereas the effect of oleic acid was similar to that of the phagostimulant saccharose. Hence, palmitic acid might be applied directly in H. obscurus control programs to prevent or reduce feeding and oviposition behaviors or might be used as a tool for selecting resistant cultivars.

El contenido de ácidos grasos de cadena larga en extractos apolares de raíces de trébol rosado de 1,6 años de edad provenientes de dos cultivares (Superqueli INIA y Quiñequeli INIA) y una línea experimental (Syn II Pre III), su relación con poblaciones de H. obscurus, y el efecto estimulante o disuasivo de extractos de raíces y ácidos grasos individuales sobre la conducta de alimentación de H. obscurus fueron investigados. Se determinó una relación inversa en el cultivar Superqueli INIA entre la población de insectos y el contenido de ácido palmítico. Cuando soluciones de ácido palmítico y oleíco fueron incorporadas individualmente en una dieta artificial, solo el ácido palmítico mostró un efecto disuasivo en la alimentación, mientras que el ácido oleico mostro un efecto similar al fagoestimulante sacarosa (control positivo). Por lo tanto, el ácido palmítico podría ser utilizado en programas de control de H. obscurus aplicado directamente para prevenir o reducir la conducta de alimentación y oviposición, o convertirse en una herramienta para la selección de cultivares resistentes a este insecto.

Background Enteric red mouth disease and Saprolegniasis, which are caused by the bacteria Yersinia ruckeri and the oomycete Saprolegnia parasitica, respectively, are important illnesses that affect salmonid farming. Sanitary problems in farms are addressed by the prevention of disease outbreaks or by the treatment of diseases with chemicals. Environmental and governmental restrictions, toxicity and high treatment costs limit the use of drugs. Marine organisms, such as algae, sponges and corals, have developed an antimicrobial defense strategy based on the production of bioactive metabolites. Among these organisms, seaweeds offer a particularly rich source of potential new drugs. Hence, many pharmacologically active substances have been isolated from seaweeds. In the Ceramium genus, Ceramium rubrum has been emphasized by several authors for its antimicrobial properties. Based on this background, the present study focused on the antimicrobial activity of a lipophilic extract of C. rubrum on Y. ruckeri and S. parasitica. Results The alga, collected from the Pacific coast of Chile, underwent an ethanol extraction, and the concentrated extract was partitioned between water and dichloromethane. From the dichloromethane extract, fatty acids, fatty acid esters, one hydrocarbon and phytol were identified by Gas Chromatography-Mass Spectrometry (GC/MS) analysis. The antimicrobial study showed that the whole extract was more active than the individual components, which suggests a strong synergistic effect among the components. Conclusions These results may constitute a basis for promising future applied research that could investigate the use of C. rubrum seaweed as a source of antimicrobial compounds against fish pathogens.

L. Parra, A. Mutis, F. Ortega, and A. Quiroz. 2013. Field response of Hylastinus obscurus Marsham (Coleoptera: Curculionidae) to ,E-2-hexenal and limonene, two host-derived semiochemicals. Cien. Inv. Agr. 40(3): 637-642. Red clover (Trifolium pratense L.), a valuable legume in Chile, is important mainly for seed production and as forage for animal production. Although it is considered a perennial, the productivity of this legume declines during the second year of growth primarily because of infestation by the red clover root borer, Hylastinus obscurus (Coleoptera: Curculionidae). Currently, there is no efficient method for controlling this insect, although the use of semiochemicals may provide a new alternative to reduce the damage caused by this curculionid. In this context, E-2-hexenal and limonene, which were previously determined to be an attractant and a repellent, respectively, for H. obscurus, were released from dispensers established in two cultivars (Quiñequeli INIA and Superqueli INIA) and one experimental line (Syn II Pre III) of red clover at the Centro Regional de Investigación Carillanca (INIA-Carillanca) (Vilcún, Chile). H. obscurus was found in significantly higher numbers on plants from areas treated with E-2-hexenal than from areas treated with limonene or in areas that were left untreated as control areas. However, this attractiveness was influenced by the red clover source such that the effect of E-2-hexenal was observed in Superqueli INIA and Syn II Pre III but not in Quiñequeli INIA. Unlike previous studies, limonene had no effect on H. obscurus. Our results provide the first evidence under field conditions for the potential use of E-2-hexenal as an attractant for the red clover root borer H. obscurus.

L. Parra, A. Mutis, F. Ortega y A. Quiroz. 2013. Respuesta de Hylastinus obscurus Marsham (Coleoptera: Curculionidae) a ,E-2-hexenal y limoneno, dos semioquímicos derivados de su hospedero, en condiciones de campo. Cien. Inv. Agr. 40(3): 637-642. El trébol rosado es una valiosa leguminosa, cuya importancia en Chile radica principalmente en la producción de semillas y su uso como alimento forrajero. A pesar de ser una planta perenne, su producción declina durante el segundo año de establecida, debido principalmente a la infestación del barrenador de la raíz del trébol Hylastinus obscurus (Coleoptera: Curculionidae). Actualmente no existe un método de control eficiente contra este insecto, por lo tanto, el uso de semioquímicos surge como una nueva alternativa para disminuir los daños ocasionados por este curculiónido. En este contexto, E-2-hexenal y limoneno, determinados previamente como atrayente y repelente a H. obscurus, fueron dispuestos en dispensadores de liberación lenta establecidas en dos cultivares y una línea experimental de trébol rosado en Centro de Investigación INIA Carillanca (Vilcún, Chile). Las evaluaciones indican que H. obscurus fue significativamente encontrado en mayor proporción en plantas sometidas al tratamiento con E-2-hexenal, comparadas con limoneno y el control. Sin embargo, este efecto atrayente fue influenciado por los cultivares y línea experimental de trébol rosado, de tal manera que el efecto de E-2-hexenal fue observado en Superqueli INIA y Syn II Pre III, pero no en Quiñequeli INIA. En contraste a previos estudios, limoneno no tuvo efecto sobre H. obscurus. Nuestros resultados permiten establecer la primera evidencia bajo condiciones de campo del potencial uso, especialmente E-2-hexenal, como un atrayente para H. obscurus.

Background: The horn fly, Haematobia irritans, is an obligate bloodsucking ectoparasite of pastured cattle and is a major pest of livestock production in North and South America and Europe. In this study, we investigated the potential to use cattle pastures, infected with non-toxic, "friendly" fungal-endophyte-infected (E+) tall fescue, Festuca arundinacea Schreb., as a strategy for reducing horn fly loads in cattle, and to evaluate the possible bioinsecticide effect on horn fly larvae. Results: When cattle grazed in E+ tall fescue, a decrease in fly-load was observed, compared with other pastures (endophyte-free (E-) pastures). The infestation of horn fly load decreased according to an increase in the percentage of endophyte present in the different pastures (0 to 100%). Moreover, two groups of animals with significant differences in the fly-load (high and low fly-load) in the same herd were observed (P < 0.05). Additionally, it was possible to determine a bioinsecticide effect of cattle dung, upon horn fly larvae (80%), from animals fed E+ tall fescue. Conclusions: These results constitute the first report on the potential for exploiting pasture management for controlling 1) horn fly-loads on cattle and 2) the normal development of horn fly larvae. In conclusion, this information provides preliminary understanding of the role of cattle pasture diet management for controlling horn fliesas part of an integrated pest management strategy for this major pest of farmed livestock.