Infection of perennial ryegrass (Lolium perenne) by an endophyte fungus (Neotyphodium lolii) decreases the abundance and diversity of predators and parasitoids

ABSTRACT Perennial ryegrass is one of the most important food sources in animal production. However, several pests affect this crop, and one of the primary control strategies is the symbiotic relationships between ryegrass endophyte fungi. This fungus produces alkaloids that exhibit toxic activity against arthropods. Furthermore, the effect of fungi may extend to higher trophic levels, including predators (spiders and/or insects), decreasing their abundance and diversity. Given the importance of spiders and insects as predators, whether the symbiotic interaction between perennial ryegrass and endophyte fungus reduces the abundance and diversity of predators pose an important question. To address this question, natural enemies in perennial ryegrass were collected and analyzed over a year, and the percentage of endophyte fungus was evaluated by the presence of hyphae from two ryegrass cultivars, Jumbo (E-) and Alto AR1 (E+). We observed an 80% endophyte infection rate for (E+) and 0% for (E-). Moreover, 222 individual spiders corresponding to 10 families were identified in both perennial ryegrasses, including 209 individuals for (E-) and 13 for (E+). The most abundant spider family was Lycosidae, representing 71.17% of the total spiders. In addition, 65 insects were collected, corresponding to 6 families, with Carabidae being the most abundant. Furthermore, the Simpson index indicated the dominance of the family Lycosidae. Overall, spider and insect abundance and diversity were reduced in (E+), suggesting a negative effect of the endophyte on predator populations.

Volatiles induction in response to mechanical damage is reduced by domestication in murtilla / La inducción de volátiles en respuesta al daño mecánico es reducida por la domesticación en murtilla

Volatiles compounds are involved in defensive induction against insects, playing an important role in insect-plant interaction being induced by response to mechanical damage. However, they could decrease according to the domestication degree in cultivated plants. Currently, it has been established that secondary metabolites are reduced due to the domestication process in murtilla. Hence, the follow question emerges: Are volatile organic compounds induced by mechanical damage reduced in cultivated murtilla plants in relation to wild plants? Two cultivated ecotypes and their respective wild counterparts were sampled. Volatiles compounds were obtained using Porapak-Q columns and analyzed by gas chromatography. Results showed that compounds as 2-hexanone, α-pinene, 2-thujene, 3-thujene and 1,8- cineole were more abundant in wild plants exposed to a mechanical damage than cultivated plants. Hence, these compounds have been associated to induced defense, these results suggest that domestication reduced the induction of defensive volatiles in cultivated murtilla in response to mechanical damage.

Los compuestos volátiles están implicados en la defensa inducida contra insectos, desempeñando un papel importante en esta interacción. Sin embargo, estos compuestos podrían disminuir según el grado de domesticación. Actualmente, se ha reportado que algunos metabolitos secundarios son reducidos en plantas de murtilla domesticadas. Por lo tanto, surge la siguiente pregunta de investigación: ¿Los compuestos orgánicos volátiles inducidos por el daño mecánico son reducidos en plantas cultivadas de murtilla en comparación con plantas silvestres? Para dos ecotipos cultivados y sus respectivas contrapartes silvestres, los compuestos volátiles fueron capturados usando columnas de Porapak-Q y las muestras analizadas por cromatografía gaseosa. Los resultados mostraron que compuestos tales como 2- hexanona, α-pineno, 2-tujeno, 3-tujeno y 1,8-cineol fueron más abundantes en plantas silvestres expuestas a daño mecánico que en cultivadas. Debido a que estos compuestos se han asociado a defensa inducida, estos resultados sugieren que la domesticación reduce la inducción de volátiles en plantas cultivadas sometidas a daño mecánico.

Differential formononetin content in cultivars and experimental lines of red clover (Trifolium pratense L. ) plants affect the feeding behaviour of Hylastinus obscurus (Coleoptera: Curculionidae) / Contenido diferencial de formononetina en cultivares y líneas experimentales de plantas de trébol rosado (Trifolium pratense L. ) afecta la conducta alimenticia de Hylastinus obscurus (Coleoptera: Curculionidae)

Red clover (Trifolium pratense L.) is a perennial plant widely used as a forage resource for several animals. This plant is the exclusive host of Hylastinus obscurus (Marsham) which causes irreparable damages to the root system affecting their persistence. It has been reported that the presence of the isoflavonoid formononetin in roots of red clover could act as an antifeedant on H. obscurus. There are not studies related to the formononetin content in red clover roots to the antifeedant effect elicited by experimental lines and cultivar of red clover. Six red clover genotypes were investigated in both formononetin content and their respective antifeedant action. The results showed to Sabtoron High and Superqueli-INIA with both the highest formononetin content in red clover roots and antifeedant effect, allowing to suggest that this secondary metabolites could be used as a chemical factor for red clover plants. Moreover, a rapid methodology for searching red clover genotypes with high formononetin content is reported.

El trébol rosado (Trifolium pratense L.) es una planta perenne ampliamente utilizada como fuente de forraje de variados animales. Esta planta es el exclusivo hospedero de Hylastinus obscurus (Marsham) el cual causa irreparables daños al sistema radical afectando seriamente su persistencia. Se ha reportado que la presencia del isoflavonoide formononetina en raíces del trébol rosado podría actuar como antialimentario sobre H. obscurus. Actualmente no existen estudios que relacionen el contenido de formononetina en raíces de trébol rosado con el efecto antialimentario elicitado por líneas experimentales y cultivares de trébol rosado. Seis genotipos de esta leguminosa fueron evaluados en cuanto a su contenido de formononetina y actividad antialimentaria. Los resultados mostraron que los cultivares Sabtoron High y Superqueli-INIA presentaron altos niveles de formononetina en sus raíces y efecto antialimentario sobre H. obscurus, lo que permite sugerir que este metabolito secundario podría ser usado como factor químico para incrementar la persistencia de plantas de trébol rosado. Además, se informa una metodología rápida para la búsqueda de genotipos con altos contenidos de formononetina.

Rhodolirium andicola: a new renewable source of alkaloids with acetylcholinesterase inhibitory activity, a study from nature to molecular docking

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.

El adipocito en el laboratorio: historia, perspectivas y reporte de experiencia / The adipocyte in the laboratory: history, perspectives and report of experience

Introducción: el tejido adiposo humano está compuesto por diferentes tipos celulares, y ha sido objeto de múltiples estudios en los últimos años debido a su acción en diversas funciones. Métodos: se realizó una búsqueda bilbiográfica en PubMed, Scielo, Science Direct and Google Scholars y se reporta la experiencia de los autores. Resultados: los primeros modelos de estudio fueron con tejido de roedores, y permitieron la comprensión del metabolismo de carbohidratos y ácidos grasos y facilitaron el estudio de la biología del adipocito, junto a estudios histológicos y el aislamiento de adipocitos maduros. Posteriormente se realizaron estudios con células precursoras (preadipocitos) que propiciaron el aislamiento de la línea celular 3T3-1L murina. En Latinoamérica, se han realizado diversos estudios con líneas celulares y con células madre mesenquimales precursoras de adipocitos para estudiar el efecto de hormonas y otras sustancias y para genotipificación. En Colombia se han realizado estudios con adipocitos 3T3-L1 para determinar los efectos de medicamentos y sustancias en estas células. En el laboratorio de Fisiología Celular de la Universidad Tecnológica de Pereira el proceso de obtención de muestras ha evidenciado dificultades por tratarse de tejido humano, pero el protocolo de aislamiento y cultivo pudo ser estandarizado a lo largo de seis años de experimentación; se aislaron preadipocitos y adipocitos maduros que permitieron estudiar los efectos de hormonas, realizar caracterización electrofisiológica y estudiar la fisiología del calcio. Conclusiones: este es un campo de investigación muy relevante debido a la implicación de este tipo celular en funciones metabólicas sistémicas y su relación con patologías de alta prevalencia como la obesidad y el síndrome metabólico.

Introduction: The human adipose tissue is composed of different cell types. It has been subject of several studies in the past years due to its role on diverse functions. Methods: A search in the PubMed, Scielo, Science direct and Google Scholars databases was performed and the authors experience was reported. Results: The first model used was rodent fat, which allowed a better comprehension on carbohydrate and fatty acid metabolism and enhanced research in adipocyte cell biology in addition with histological studies and mature adipocyte isolation. Afterwards, learning about precursor cell (pre-adipocytes) promoted the isolation of murine 3T3-L1 cell line. In Latin America research has been conducted using cell lines and adipocyte precursor mesenchymal stem cells to describe effects of hormones and perform DNA sequencing. In Colombia, studies in 3T3-L1 cell line aimed to stablish the effects of different compounds on these cells. In the Cell Physiology Laboratory of the Universidad Tecnológica de Pereira, sample collection process has shown difficulties because the source was human tissue; nevertheless isolation and cell culture protocols were standardized throughout the last six years of experimentation. Pre-adipocytes and mature adipocytes were isolated to study the effects of hormones, perform electrophysiological characterization and study calcium physiology. Conclusions: This is a relevant research field since these cells have important systemic metabolic functions and they have a clear relationship with high-prevalence pathologies such as obesity and the metabolic syndrome.

A rapid preparative-TLC/GC-MS methodology for discriminating between two filmy ferns (Hymenophyllaceae) native from the temperate rain forest of Southern Chile based on their soluble carbohydrates / Una metodología de CCF-preparativa/CG-EM rápida para discriminar entre dos helechos película(Hymenophyllaceae) nativos del bosque templado del sur de Chile basada en sus hidratos de carbono

Filmy ferns are a conspicuous epiphytic component of the temperate rain forest of southern Chile and the more abundant genus is Hymenophyllum represented by 19 species, they are a primitive fern family sharing their limited water lost control (poikilohydricity) with lower non-vascular plants, such as bryophytes and lichens. Because, carbohydrate accumulation is a desiccation tolerance mechanism proposed for desiccation-tolerant vascularized plants, the aim of this investigation was to propose a methodology for soluble carbohydrate analysis in native ferns. Preparative- TLC showed to be a simple and rapid technique for separating Hymenophyllum sugars. GC-MS analysis allowed identifying glucitol (2.0 percent), sucrose (2.0 percent), alpha or beta-melibiose (5.0 percent) and salidroside (4.4 percent) in H. cruentum carbohydrate profile from H. dentatum showed significant differences presenting only two sugars sucrose (1.4 percent) and alpha or beta-melibiose (3.2 percent). Furthermore, salidroside, an antioxidant compound, was identified only in H. cruentum. Our results suggest that H. cruentum, has higher preventive mechanisms than H. dentatum; that would allow delay the effects of desiccation (prevent a rapid dehydration) and thus survive to short periods of drought present in the study area. Due to the presence of salidroside in H. cruentum this species should be more resistant to oxidative stress that H. dentatum.

Los helechos película se pueden encontrar en el bosque templado lluvioso del sur de Chile, siendo el género más abundante Hymenophyllum representado con 19 especies. Son una familia de helechos primitivos que comparten la limitación de control de pérdida de agua (poiquilohidrícos) con plantas no vasculares, tales como briófitas y líquenes. Como el contenido de hidratos de carbono ha sido asociado a la protección de la planta durante la desecación, su análisis es una herramienta importante para dilucidar el mecanismo asociado a este fenómeno. El objetivo de esta investigación fue proponer una metodología fácil y rápida para el análisis de hidratos de carbono en helechos nativos. La CCF-preparativa permitió la separación de hidratos de carbono presentes en plantas del género Hymenophyllum. El análisis por GC-MS logró identificar glucitol (2,0 por ciento), sacarosa (2,0 por ciento), alfa o Beta-melibiosa (5.0 por ciento) y salidrosido (4,4 por ciento) en H. cruentum. El perfil de hidratos de carbono de H. dentatum mostró diferencias significativas, mostrando sólo dos compuestos sacarosa (1,4 por ciento) y alfa o beta- melibiosa (3.2 por ciento). Además, salidrosido, un compuesto antioxidante, sólo fue identificado en H. cruentum. Nuestros resultados sugieren que ante la menor tolerancia a la desecación observada previamente en H. cruentum, esta especie presenta mecanismos que le permitirían retardar los efectos de la desecación (prevención de una rápida pérdida de agua) y así sobrevivir a los periodos cortos de sequía presentes en la zona de estudio. Debido a La presencia de salidrosido en H. cruentum, esta especie debería ser más resistente al estrés oxidativo que H. dentatum.

Identification of isoquinoline alkaloids from Berberis microphylla by HPLC ESI-MS/MS / Identificación de alcaloides isoquinolínicos en Berberis microphylla G. Forst mediante CLAE IES-MS/MS

Berberis microphylla (G. Forst) is a native plant growing in Patagonia. In recent years Patagonia Berberis are becoming important due to their interesting biological properties related to their alkaloids content. The aim of this study was determine the distribution and proportion of isoquinoline alkaloids in leaves, stems and roots of B. microphylla collected in two different climatic zones from Chilean Patagonia. Using by HPLC ESI-MS/MS isocorydine, jatrorrhizine, palmatine, reticuline, scoulerine, tetrahydroberberine and thalifendine were detected for the first time in this specie, and the presence of allocryptopine, berberine, calafatine and protopine, previously isolated in B. microphylla was corroborated. The alkaloids profile showed differences of compounds in samples collected in two climatic zones, where more compounds were detected in plants from Lago Deseado than Cerro Sombrero. Furthermore, a greater number of alkaloids were found in stem and root extracts and berberine and thalifendine were detected in higher proportion in these structures.

Berberis microphylla (G. Forst) es un arbusto nativo que crece en la Patagonia. Actualmente, esta planta ha sido foco de estudio dada las propiedades biológicas que presenta, atribuidas principalmente al contenido de alcaloides. El objetivo de este estudio fue determinar la distribución y proporción de alcaloides isoquinolínicos en hojas, tallos y raíces de B. microphylla colectadas en dos zonas climáticas de la Patagonia chilena. Mediante CLAE IES-MS/MS se informa por primera vez la presencia de isocoridina jatrorrizina, palmatina, reticulina, escoulerina, tetrahidroberberina y talifendina en esta especie y se confirma la presencia de allocriptopina, berberina, calafatina y protopina, identificados previamente en B. microphylla. El perfil de alcaloides mostró diferencias en la presencia de compuestos en las muestras colectas en las dos zonas climáticas, observándose un mayor número de compuestos en plantas provenientes de Lago Deseado. Además, un mayor número de compuestos se identificó en extractos de tallos y raíces donde berberina y talifendina fueron detectados en mayor proporción.

Novel antimicrobial activity of a dichloromethane extract obtained from red seaweed Ceramium rubrum (Hudson) (Rhodophyta: Florideophyceae) against Yersinia ruckeri and Saprolegnia parasitica, agents that cause diseases in salmonids

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.

Chemical composition and antibacterial activity of Laureliopsis philippiana (Looser) essential oil / Composición química y actividad antibacteriana del aceite esencial de Laureliopsis philippiana (Looser)

Laureliopsis philippiana (Looser) is native evergreen specie from Chile and Argentina used in traditional medicine. In this study, chemical composition as well as its in vitro antibacterial activity against Escherichia coli, Enterobacter aerogenes, Staphylococcus epidermidis and Staphylococcus aureus of essential oil from leaves of this species was determinated. Chemical analysis by GC-MS resulted in the identification of 19 compounds representing 98.8 percent onoterpenes; linalool (32.3 percent) and eucalyptol (37.4 percent) were the main constituents. To evaluate the antibacterial activity disc diffusion method and broth dilution method were used. The essential oil exhibited inhibitory activity against Gram (-) and Gram (+) bacteria, whereas similar activity to essential oil was showed for linalool against E. aerogenes and S. epidermidis whereas linalool alone, achieves an inhibitory effect against E. aerogenes and S. epidermidis comparable to the essential oil.

Laureliopsis philippiana (Looser) es una especie siempre verde nativa de Chile y Argentina usado en medicina tradicional. En este estudio se determinó tanto la composición química del aceite esencial obtenido a partir de hojas de esta especie, así como su actividad antibacterial in vitro contra Escherichia coli, Enterobacter aerogenes, Staphylococcus epidermidis and Staphylococcus aureus. El análisis químico por GC-MS permitió la identificación de 19 compuestos, representando el 98,8 por ciento de la composición del aceite. Monoterpenos oxigenados, eucaliptol y linalol fueron los mayores constituyentes del aceite con un 37,4 por ciento y 32,3 por ciento respectivamente. Para evaluar la actividad antibacteriana se utilizaron los métodos de difusión en agar y dilución en caldo. El aceite esencial muestra actividad inhibitoria contra las bacterias Gram (-) y Gram (+) evaluadas, mientras que linalol por si solo logra un efecto inhibitorio comparable con el aceite esencial contra E. aerogenes y S. epidermidis mientras que el linalol por si solo, logra un efecto inhibitorio contra E. aerogenes y S. epidermidis comparable al del aceite esencial.

Differences in the fly-load of Haematobia irritans (Diptera: Muscidae) on cattle is modified by endophyte infection of pastures

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.

Asociación entre prácticas en salud, somatización y algunos factores psicosociales en usuarios de clínicas de ASSBASALUD ESE: Manizales (Colombia), 2007 / Association between health practices, somatization and some psychosocial factors in clinics of ASSBASALUD ESE: Manizales (Colombia), 2007

Introducción: La evidencia investigativa sugiere que la población con una patología médica crónica presenta mayor riesgo de desarrollar un cuadro de somatización, definida como una serie de síntomas corporales sin causa orgánica demostrable, y asociada al estrés y cuadros depresivos, ansiosos, que generalmente pasan desapercibidos en la consulta. Este tipo de pacientes terminan convirtiéndose en consultantes crónicos o en abusadores de servicios de salud, lo que genera congestión en la prestación del servicio e incremento en los costos de salud. Metodología: Estudio de corte transversal, con 188 pacientes que asistieron a consultaexterna en los diferentes centros urbanos de Atención en Seguridad Social, Bienestar y Salud, Empresa Social del Estado (ASSBASALUD, ESE) del área urbana de Manizales (Colombia). Resultados: La edad promedio fue 51,3 años, 62,5 por ciento presentaban somatización, 61,9 por ciento ansiedad subclínica, 51,6 por ciento no presentaban depresión, estrés bajo 73,6 por ciento y buena función familiar 42,9 por ciento . Se encontró dependencia significativa entre somatización y ansiedad (p=0,000), depresión (p=0,005) estrés (p=0,000) funcionalidad familiar (p=0,007), entre otras. También se encontró dependencia significativa entre las variables prácticas en salud y somatización (p=0.002), Ansiedad (p=0,014), depresión (p=0.008) a nivel p<0,1 con funcionalidad familiar (p=0,093). Conclusiones: se encontró una alta comorbilidad de somatización en los pacientes con algún grado de depresión, ansiedad y estrés, al igual que deficientes prácticas de salud en este tipo de pacientes.