Microsporogénesis y ultraestructura de los granos de polen en la planta del cacao, Theobroma cacao (Malvaceae)

Introducción: No conocemos estudios sobre la microsporogénesis de la planta de cacao, y poco se sabe sobre la ultraestructura de sus granos de polen. Objetivo: Describir la microsporogénesis y ultraestructura de los granos de polen en T. cacao. Métodos: Procesamos más de 30 flores para cada etapa floral, teñidas con Safranina-Azul Alcian, PAS-Amidoblack y Lacmoid. Para la microscopía de transmisión procesamos las muestras en resina y las teñimos con azul de toluidina. Para microscopía electrónica de barrido, fijamos y deshidratamos en 2.2-dimetoxipropano, secamos hasta un punto crítico y recubrimos con oro. Resultados: Anteras diferenciadas por una masa celular en los extremos distales a los filamentos estaminales. Durante el desarrollo la pared de las anteras presenta varios estratos celulares y al madurar se reducen a la epidermis y al endotecio. Las células madre de microsporas se dividen por meiosis para formar tétradas. El tapete es secretor e intacto hasta que se liberan los granos, para luego degenerar. Los granos de polen son isopolares, esferoidales, pequeños, tricolpados. La ultraestructura presenta una esporodermis semitectada, con ornamentación reticulada, y un retículo heterobrochado con el muri sin ornamentación. La exina se deposita antes que la intina. Los orbículos son individuales, lisos y de tamaño variado. Hay abundante polenkit en el tectum y entre las columelas. La intina es delgada, pero se desarrolla ampliamente en las áreas del colpo, formando una intina interna compacta y una intina externa inusual con una apariencia columelada. Conclusión: La estructura y el desarrollo de las anteras siguen el patrón de las angiospermas. La microsporogénesis simultánea y la deposición centrípeta de la esporodermis se conocen de Malvaceae, pero los caracteres de la intina son nuevos para la familia.

Introduction: We know of no studies on the microsporogenesis of the cocoa plant, and little is known about the ultrastructure of its pollen grains. Objective: To describe microsporogenesis and ultrastructure of pollen grains in T. cacao. Methods: We processed over 30 flowers for each floral stage and stained with Safranin-Alcian Blue, PAS-Amidoblack and Lacmoid. For transmission microscopy we processed samples on resin and stained with toluidine blue. For scanning electron microscopy, we fixed and dehydrated in 2.2-dimethoxypropane, critically dried and coated with gold. Results: Anthers differentiated by a cellular mass at the ends distal to the staminal filaments. During development, the anther wall has several cellular layers reduced, at maturity, to the epidermis and endothecium. Microspore mother cells divide by meiosis to form tetrads. The tapetum is secretory and intact until the grains are released, to later degenerate. Pollen grains are isopolar, spheroidal, small, tricolpate. Ultrastructure has a semi-tectate sporodermis, with reticulate ornamentation, and heterobrochated reticulum with the muri without ornamentation. Exine is deposited before intine. The orbicles are individual, smooth, and varied in size. There is abundant pollenkitt on the tectum and between the columellae. The intine is thin, but develops widely in the colpus areas, forming a compact internal intine and an unusual external intine with a columellated appearance. Conclusion: Anther structure and development follows the angiosperm pattern. Simultaneous microsporogenesis and centripetal deposition of the sporodermis are known from Malvaceae, but intine characters are novel for the family.

Caracterización física y sensorial de 24 genotipos especiales de cacao Theobroma cacao

Colombia, se destaca como país productor de cacao fino y de aroma, por lo cual, resulta de gran importancia explorar las características físicas y sensoriales de la diversidad genética existente. Para el presente estudio, se seleccionaron 22 genotipos de colecciones de trabajo con atributos productivos sobresalientes y dos testigos comerciales, reconocidos por su productividad y calidad sensorial; las muestras evaluadas fueron tomadas, de acuerdo con un muestreo aleatorio simple y se realizó un proceso de poscosecha homogéneo, controlado e independiente por genotipo, para obtener muestras de cacao seco individuales. Se realizaron análisis físicos al grano de cada genotipo y se preparó licor de cacao con cada una de las muestras obtenidas. Posteriormente, se evaluó cada licor por medio del panel de evaluación sensorial entrenado, donde se identificó y cuantificó la intensidad de atributos básicos, atributos especiales y atributos adquiridos, característicos de cada muestra. La información fue analizada mediante estadística descriptiva y análisis de componentes principales, lo que permitió la identificación de tres genotipos sobresalientes por el índice de grano y el porcentaje de cascarilla. El análisis sensorial evidenció que, en diecisiete genotipos, predominaron los atributos especiales, como herbal, floral, frutal, frutos secos y dulce. Estos resultados son un aporte importante para la selección de nuevas variedades de alta productividad, con características sensoriales de interés para la comercialización, que pueden ser evaluadas en diferentes regiones, para aumentar la disponibilidad genética en futuros programas de renovación y siembra de cacao que está en expansión, en los distintos territorios del país.

Colombia stands out as a country that produces fine and aroma cocoa; therefore, it is of great importance to explore the physical and sensory characteristics of the existing genetic diversity. For the present study, 22 genotypes from working collections with outstanding productive attributes and two recognized commercial controls were selected and evaluated for its productivity and sensory quality; the evaluated samples were taken according to a simple random sampling and a homogeneous, controlled, and independent post-harvest process was carried out by genotype, to obtain individual dry cocoa samples. Physical analyzes were carried out on the grain of each genotype and cocoa liquor was prepared with each of the samples obtained. Subsequently, each liquor was evaluated by means of the trained sensory evaluation panel, where the intensity of basic attributes, special attributes and acquired attributes characteristic of each sample was identified and quantified. The information was analyzed through descriptive statistics and principal component analysis, which allowed the identification of three outstanding genotypes by the grain index, and the husk percentage. The sensory analysis showed that, in seventeen genotypes, special attributes predominated such as: herbal, floral, fruity, dried fruit and sweet. These results are an important contribution for the selection of new high productivity varieties with sensory characteristics of interest for commercialization, which can be evaluated in different regions, to increase genetic availability in future renewal and planting programs of cocoa that is expanding in the different territories of the country.

Genetic variability of cocoa from the amazon basin: implications for genetic conservation

The cocoa and chocolate production chain involves US$60 billion annually and three million farmers around the world, in an area exceeding nine million hectares. The use of wild germplasm will enable to generate new disease- and pest-resistant cultivars and ability to adapt to changing environments. Here we evaluated 145 cocoa accessions, originated from nine Amazonian basins, based on eight fruit traits. Univariate anova showed significant differences (p<0.05) for all traits. For seven traits, the variance component within basins was higher (81.5%, on average). Therefore, it is recommended that the collection of wild accessions prioritize a larger number of plants from a few populations of the most divergent basins. The multivariate analyses revealed a greater divergence between the Ji-Paraná-RO and Solimões/Amazonas-PA basins (27.69) and a greater similarity between Alien clones-PER and Solimões/Amazonas-AM (0.66) in relation to their populations. They also revealed that the accessions differentiation occurred according to the river basin system. These results allowed elucidate the genetic structure and distribution of cacao populations. In addition, strengthen the importance of collecting and conserving germplasm to preserve genetic resources.

Obtención y caracterización fisicoquímica de un sustrato nutritivo hecho a partir de cáscaras de cacao para la producción de plántulas de hortalizas / Obtaining and physicochemical characterization of a nutritive substrate made from cacao shells for the production of vegetable seedlings

En el beneficiado del cacao se producen una gran cantidad de residuos, las mazorcas generalmente son desechadas dentro de los mismos cultivos y genera problemáticas como la proliferación de insectos y microorganismos patógenos. De estos desechos las cáscaras son las de mayor relevancia. La Escuela de Ingeniería Química de ITCA-FEPADE evaluó añadirle valor a este subproducto, incorporando desechos de cáscara de cacao en la formulación de un sustrato para el cultivo de plántulas de hortalizas. El objetivo de esta investigación fue obtener un sustrato orgánico que sirva de soporte material y nutritivo a partir de cáscaras de cacao criollo. La biomasa vegetal se caracterizó teniendo en cuenta parámetros como el porcentaje de humedad, pH, porcentaje de cenizas, contenido de potasio, nitrógeno y fósforo. Se ejecutaron pruebas comparativas de formulación del sustrato, siembra, cultivo y crecimiento de las plántulas de hortalizas de tomate y pepino, obteniendo como resultado la fórmula óptima de un sustrato y abono orgánico y el mejor medio de desarrollo. De los resultados obtenidos se concluye que, a partir de un adecuado procesamiento y aprovechamiento de las cáscaras de cacao, se puede transformar este desecho en un producto biomaterial alternativo que genera una opción de bioprospección agroindustrial. Los valores obtenidos en la caracterización fisicoquímica de las cáscaras de las mazorcas, dependen de condiciones como el tipo de suelo, variables agrometeorológicas, calidad de agua, abono y especie de la planta de cacao. El escenario de siembra condiciona el desarrollo óptimo de las plántulas; los factores como requerimiento de agua, distribución de nutrientes y estabilidad de las plántulas, se ven afectados por la relación del espacio de germinación. Como resultado de la caracterización fisicoquímica, se obtuvo: pH de 5.7, cenizas 18.83%, humedad 73.56%, celulosa 21.39%, lignina 39.81%, nitrógeno total 0.02%, fósforo total 0.02% y ausencia de potasio.

In the cacao beneficiation, a large amount of waste is produced, generally, the cacao pods are discarded within the same crops and generate problems such as the proliferation of insects and pathogenic microorganisms. The shell are the most relevant this waste. In Escuela de Ingeniería Química of ITCA-FEPADE the incorporation of cacao shell waste in the formulation of a substrate for growing vegetables was evaluated because the giving benefit to these by-products. The objective of this research was to obtain a substrate that serves as material and nutritional support for vegetable seedlings, from creole cacao shells. The vegetal biomass was characterized taking into account parameters such as moisture percentage, pH, ash percentage, potassium, nitrogen and phosphorus content. Subsequently, comparative tests of formulation, planting and growth of vegetable tomato and cucumber seedlings were carry out, obtaining as result, the optimal formula for the substrate and the best development environment. It concludes that, through an adequate processing and use of the cacao shell, it is possible to transform a waste into an alternative biomaterial product that generates an agro industrial bioprospecting option. The values obtained in the physicochemical characterization of the shells of cacao pods depend on conditions such as soil type, agrometeorological variables, water quality, fertilizer and cacao plant species. The planting environment determines the optimal development of the seedlings, factors such as water requirement, nutrient distribution and seedling stability seems affected because relation of the germination space. The results of the characterization were a pH of 5.7, ashes 18.83%, humidity 73.56%, cellulose 21.39%, lignin 39.81%, total nitrogen 0.02%, total phosphorus 0.02%, and total phosphorus 0.02% and an absence of potassium.

Utilización de los recursos genéticos en un país que es centro de origen: el caso del cacao en el Perú

La producción peruana de cacao representa aproximadamente el 2% de la producción mundial; con una contribución muy reconocida por la calidad y diversidad de sus sabores y aromas. De milenario origen amazónico y probada utilización de hace por lo menos 5200 años, este cultivo recién ha sido visible para el desarrollo agrario en las dos últimas décadas, cuando se le ha utilizado con relativo éxito en la sustitución de cultivos ilegales de coca ¿Y qué tanto está contribuyendo la academia a la cadena de valor del cacao? Con carácter exploratorio, se registró y analizó una muestra de 44 tesis que tenían como objeto de estudio el cacao, obtenido por sus autores en los centros de producción y parcelas de productores. La mayor parte de estas tesis (41) estuvieron destinadas a obtener títulos profesionales, dos para optar el grado de magister y uno para obtener el grado de doctorado. Estas tesis se realizaron en universidades de 13 departamentos del Perú, contando Lima con el mayor número de tesis (12). Once tesis abarcaron actividades de investigación del germoplasma para la transformación y/o la innovación del cacao, confiriéndole así valor agregado con potencial uso comercial. Estas investigaciones estuvieron relacionadas con cuatro sectores productivos con un claro potencial comercial en el mercado global de recursos genéticos. En ningún caso, las investigaciones se realizaron bajo la seguridad jurídica del sistema de acceso a recursos genéticos y participación justa y equitativa de beneficios del Protocolo de Nagoya, del Convenio sobre la Diversidad Biológica.

The Peruvian production of cocoa represents, approximately, 2% of the world production, with a very recognized contribution because of the quality and diversity of its flavors and aromas. With a millenary Amazonian origin and proved utilization by at least 5200 years ago, this crop just became visible for the agrarian development in the last two decades, when it was used with relative success for the substitution of illegal coca crops. And how much is the academy contributing to the value chain of cocoa? A sample of 44 theses which cocoa obtained from production centers and farmers plots was their object of study, has been registered and analyzed in an exploratory way. 41 theses were to get professional titles, two for magister degree and one for doctorate degree. These theses were done in universities of 13 Department of Peru, where Lima reached the highest number (12). Eleven theses addressed germplasm research activities aimed to cocoa transformation and/or innovation, thus conferring added value with potential commercial use. These researches were related to four productive sectors with a clear potential in the global market of genetic resources. In any case, the studies were done under the legal certainty of the system of access to genetic resources and fair and equitable benefit-sharing of the Nagoya Protocol to the Convention on Biological Diversity.

Obtención y caracterización físico química de un sustrato nutritivo hecho a partir de los desechos del cacao para la producción de plántulas de hortalizas: en asocio con empresa Chukwa Chocolates / Obtaining and physicochemical characterization of a nutritive substrate made from cacao waste for the production of vegetable seedlings: in association with Chukwa Chocolates

En el beneficiado del cacao se producen una gran cantidad de residuos, las mazorcas generalmente son desechadas dentro de los mismos cultivos y genera problemáticas como la proliferación de insectos y microorganismos patógenos. De estos desechos las cáscaras son las de mayor relevancia. La Escuela de Ingeniería Química de ITCA-FEPADE evaluó añadirle valor a este subproducto, incorporando desechos de cáscara de cacao en la formulación de un sustrato para el cultivo de plántulas de hortalizas. El objetivo de esta investigación fue obtener un sustrato orgánico que sirva de soporte material y nutritivo a partir de cáscaras de cacao criollo. La biomasa vegetal se caracterizó teniendo en cuenta parámetros como el porcentaje de humedad, pH, porcentaje de cenizas, contenido de potasio, nitrógeno y fósforo. Se ejecutaron pruebas comparativas de formulación del sustrato, siembra, cultivo y crecimiento de las plántulas de hortalizas de tomate y pepino, obteniendo como resultado la fórmula óptima de un sustrato y abono orgánico y el mejor medio de desarrollo. De los resultados obtenidos se concluye que, a partir de un adecuado procesamiento y aprovechamiento de las cáscaras de cacao, se puede transformar este desecho en un producto biomaterial alternativo que genera una opción de bioprospección agroindustrial.

In the cacao beneficiation, a large amount of waste is produced, generally, the cacao pods are discarded within the same crops and generate problems such as the proliferation of insects and pathogenic microorganisms. The shell are the most relevant this waste. In Escuela de Ingeniería Química of ITCA-FEPADE the incorporation of cacao shell waste in the formulation of a substrate for growing vegetables was evaluated because the giving benefit to these by-products. The objective of this research was to obtain a substrate that serves as material and nutritional support for vegetable seedlings, from creole cacao shells. The vegetal biomass was characterized taking into account parameters such as moisture percentage, pH, ash percentage, potassium, nitrogen and phosphorus content. Subsequently, comparative tests of formulation, planting and growth of vegetable tomato and cucumber seedlings were carry out, obtaining as result, the optimal formula for the substrate and the best development environment. It concludes that, through an adequate processing and use of the cacao shell, it is possible to transform a waste into an alternative biomaterial product that generates an agro industrial bioprospecting option.

Effect of organic fertilization and pruning on cacao yield in Cundinamarca, Colombia / Efecto de la fertilización orgánica y la poda sobre la producción de cacao en Cundinamarca, Colombia

ABSTRACT Cacao has high social importance in Colombia but requires cultivation techniques that improve its productivity, especially in the production of organic cacao. Pruning and organic fertilizers were assessed as treatments to increase the bean yield in an old cacao crop in Cundinamarca. The positive effects of organic fertilizer application and pruning on the pod morphometry and bean yield were established. The application of organic fertilizers with pruning, in general, increased the length and circumference of cacao pods. A negative correlation (R=-0.618) was found among the relative chlorophyll contents in leaves at the end of the harvest period and with the number of pods per tree. A positive correlation (R=0.748) was obtained between the fresh weight of pods and the dry weight of beans. No correlation was established between the relative chlorophyll contents in leaves and the bean dry weight. Pruning plus organic fertilization increased the total dry weight of beans per tree at almost 15 %, which makes pruning with organic fertilizer application the best tool for increasing yield in organic cacao production in old plantations in Cundinamarca.

RESUMEN El cultivo de cacao tiene una alta importancia social en Colombia, pero requiere la implementación de técnicas de cultivo que mejoren su productividad, principalmente, en la producción de cacao orgánico. La poda y los fertilizantes orgánicos fueron evaluados, como los tratamientos para incrementar el rendimiento de cacao, en una plantación antigua en el departamento Cundinamarca. La aplicación de fertilizantes orgánicos y la poda tuvieron efectos positivos sobre la morfometría del fruto y el rendimiento del cacao. La aplicación de fertilizantes orgánicos con poda, en general, aumentó la longitud y el diámetro de frutos de cacao. Se encontró una correlación negativa (R=-0,618) entre el contenido relativo de clorofila en hojas al final del período de cosecha y el número de frutos por árbol. Se obtuvo una correlación positiva (R=0,748) entre el peso fresco de frutos y el peso seco de semillas. No se estableció correlación entre el contenido relativo de clorofila en hojas y el peso seco de semillas. La poda más fertilización orgánica aumentó el peso seco total de semillas por árbol, en casi un 15 %, lo que convierte la poda con aplicación de fertilizante orgánicos en una herramienta para aumentar el rendimiento de cacao orgánico en antiguas plantaciones, en el departamento Cundinamarca.

Proximal chemical evaluation, antioxidant content and fatty acids in fermented and dried cocoa beans, roasted cocoa beans and cocoa pulp bar (Theobroma cacao L. Criollo cultivar)

ABSTRACT The objective of this work was to evaluate the proximal chemical composition, antioxidant activity assays with total phenol, 2,2-Diphenyl-1-Picrylhydrazyl, 2'2-azino-bis(3-ethylbenzothiazolin-6-sulfonic acid) diammonium salt, iron reducing antioxidant power, and the fatty acid composition of fermented and dried cocoa beans, roasted cocoa beans and cocoa pulp bar. The fermented and dried cocoa beans were purchased, roasted (115 °C for 5 min) and homogenized. The mixture was brought to its melting point (58 °C for 5 min). In the cocoa pulp bar there was an increase in lipids (53.0%). The roasted affected the concentration of ashes (3.38-2.98%). Total phenol (1.8, 1.5 and 1.3 mg AEG kg-1), 2,2-Diphenyl-1-Picrylhydrazyl (65.8, 64.4 and 65.0 μmol ET kg-1), 2'2-azino-bis(3-ethylbenzothiazolin-6-sulfonic acid) diammonium salt (3.60, 3.00 and 2.30 μmol EAC kg-1) and iron reducing antioxidant power (0.09, 0.07 and 0.06 mg Fe2+ kg-1) presented significant differences (p<0.05) in the fermented and dried cocoa beans, roasted cocoa beans and cocoa pulp bar respectively, however, the changes were minimal. The cocoa pulp bar had a higher concentration of unsaturated fatty acid (89.2%), where oleic (48.6%) was predominated. Therefore, the cocoa pulp bar is a food with an important antioxidant content, moreover, it has a higher concentration of unsaturated fatty acid, which is desirable in the diet.

RESUMEN El objetivo de este trabajo fue evaluar la composición química proximal, el contenido de antioxidantes con los ensayos de fenoles totales, 2,2-Difenil-1-Picrilhidrazil, Ácido 2'2-azino-bis (3-etilbenzotiazolin-6-sulfónico), sal de diamonio y poder antioxidante reductor del hierro, y la composición de ácidos grasos en los granos de cacao fermentados y secos, granos de cacao tostados y barra de pulpa de cacao. Se compraron los granos de cacao fermentado y seco y se tostaron (115 °C durante 5 min), y se homogeneizaron. La mezcla se llevó a su punto de fusión (58 °C durante 5 min). En la barra de pulpa de cacao hubo un aumento de lípidos (53,0%). El tostado afectó la concentración de cenizas (3,38 - 2,98%). En los resultados de fenoles totales (1,80, 1,50 y 1,30 mg AEG kg-1), 2,2-Difenil-1-Picrilhidrazil (65,8, 64,4 y 65,0 μmol ET kg-1), Ácido 2'2-azino-bis (3-etilbenzotiazolin-6-sulfónico), sal de diamonio (3,6, 3,0 y 2,3 μmol EAC kg-1) y poder antioxidante reductor del hierro (0.09, 0.07 y 0.06 mg Fe2+ kg-1) presentaron diferencias significativas (p<0.05) en los granos de cacao fermentados y secos, granos de cacao tostados y barra de pulpa de cacao respectivamente, sin embargo, los cambios fueron mínimos. La barra de pulpa de cacao presentó una mayor concentración de ácidos grasos insaturados (89%), donde predominó el oleico (48,6%). Por lo tanto, la barra de pulpa de cacao es un alimento con importante contenido en antioxidantes, además, tiene una mayor concentración de ácidos grasos insaturados, lo que es deseable en la dieta.

Micromorfología y ultraestructura de las anteras y los granos de polen en diez genotipos élite de Theobroma cacao (Malvaceae) / Micromorphology and ultrastructure of anthers and pollen grains in ten elite genotypes of Theobroma cacao (Malvaceae)

Resumen Introducción: A pesar de que T. cacao es una especie importante a nivel mundial por la producción de cacao, es poco lo que se conoce sobre la micromorfología y estructura de las anteras y los granos de polen. Objetivos: Describir y analizar la estructura y micromorfología de las anteras y los granos de polen de 10 genotipos élite de esta importante especie tropical. Métodos: Se tomaron más de 30 anteras de flores en antesis de los 10 genotipos élite de T. cacao del banco de germoplasma ex situ del Centro de Investigaciones Suiza-Agrosavia (Rionegro, Santander-Colombia). El material se procesó de acuerdo con los protocolos estándar para embeber y seccionar en parafina. Las secciones obtenidas (3 μm) se tiñeron con azul de Safranina-Alcian para discriminar estructuras con paredes primarias y secundarias y polifenoles totales. Además, se usó la técnica PAS-Amidoblack para diferenciar entre polisacáridos estructurales y de reserva, así como proteínas. Para la determinación de esporopolenina y polifenoles se usó la tinción azul de toluidina y finalmente para descripciones adicionales se aplicó la tinción azul alcián-PAS-hematoxilina. Las observaciones se realizaron mediante microscopio fotónico y microscopio de epifluorescencia. Para la observación con microscopía electrónica de barrido (MEB), las anteras con los granos de polen se fijaron y deshidrataron en 2.2 dimetoxipropano, luego se desecaron hasta un punto crítico y finalmente se recubrieron con oro. Resultados: Las anteras son bitecas y están sostenidas por un largo filamento formado por un estrato epidérmico, tejido parenquimatoso y un haz vascular. La dehiscencia ocurre longitudinalmente a través del estomio. La pared de la antera madura está formada por una capa epidérmica monoestratificada, una capa de células endoteliales con engrosamientos fibrilares lignificados y se pueden apreciar restos celulares del tapete y abundantes orbículas recubriendo la cavidad de los microesporangios. Los tejidos epidérmicos y parenquimatosos de las anteras almacenan polifenoles. Las orbículas son generalmente esféricas, psiladas y exhiben las mismas reacciones de tinción y fluorescencia que la exina de los granos de polen. Los granos de polen son mónades, isopolares, pequeños (16-19 µm) con amb circular, esferoidales, tricolpados con colpos medianos o cortos (5-10 µm) con membrana ornamentada, semitectatos, reticulados, heterobrochados, las paredes del retículo ornamentadas o no, con microgránulos de diferente tamaño o escabrados. Los análisis estadísticos mostraron que existen diferencias significativas en el tamaño de los granos de polen (P ˂ 0.05). Se observa que los granos de polen más pequeños son los del genotipo TCS 19 (16.890 µm) y se diferencian del resto de genotipos, y entre estos no se observan diferencias significativas. Solo dos genotipos (SCC 19 y SCA 6) presentaron polenkit y solo uno tiene paredes perforadas (SCA 6). Conclusiones: La estructura y micromorfología de las anteras de T. cacao son similares a las descritas para otras Malvaceae. Así mismo, los granos de polen mostraron variaciones de tamaño, ornamentación de las paredes y del lumen del retículo y presencia de polenkit. Sin embargo, no se observó relación entre las variaciones de los caracteres micromorfológicos analizados en los granos de polen y los modelos de compatibilidad polínica reportados para estos genotipos.

Abstract Introduction: Despite the fact that T. cacao is an important species worldwide for cocoa production, little is known about the micromorphology and structure of anthers and pollen grains. Objectives: To describe and analyze the structure and micromorphology of the anthers and pollen grains of 10 elite genotypes of this important tropical species. Methods: More than 30 anthers of flowers in anthesis were taken of the 10 elite genotypes of T. cacao from the ex situ germplasm bank of the Suiza-Agrosavia Research Center (Rionegro, Santander-Colombia). The anthers with the pollen grains were fixated and processed according to the standard protocols for embedding and sectioning in paraffin. Sections obtained (3 μm thick) were stained with Safranin-Alcian blue to discriminate structures with primary and secondary walls and total polyphenols. Additionally, the samples were also stained with the PAS-Amidoblack technique was used to differentiate between structural and reserve polysaccharides as well as proteins. Toluidine blue staining was used for the determination of sporopollenin and polyphenols and finally Alcian blue-PAS-Hematoxylin staining was applied for additional descriptions. Observations were made using photonic microscopy and epifluorescence microscopy. For observation with scanning electron microscopy (SEM) the anthers with the pollen grains were fixed and dehydrated in 2.2 Dimethoxypropane, then desiccated to critical point and finally coated with gold. Results: The anthers are dithecal and supported by a long filament made up of an epidermal stratum, parenchymal tissue, and a vascular bundle. The dehiscence occurs longitudinally through the stomium. The anther wall is made up of a monostratified epidermal layer, followed by a layer of endothecial cells with lignified fibrillar thickenings, cellular remnants of tapetum and abundant orbicules can be seen covering the cavity of the microsporangia. The epidermal and parenchymal tissues of the anthers are abundant in polyphenols. Orbicules are generally spherical, psilated, and these exhibit the same staining and fluorescence reactions as exine from pollen grains. The pollen grains are monades, isopolar, small (16-19 µm) with circular amb, spheroidal, tricolpate with medium or short colpi (5-10 µm) with sculptured membrane, semitectate, reticulated, heterobrochate, sculptured or non- sculptured walls, with microgranules of different size or scabrate. The statistical analyzes showed that there are significant differences in the size of the pollen grains (P ˂ 0.05). It is observed that the smallest pollen grains are those of the TCS 19 genotype (16.890 µm) and are different from the other genotypes, and among these there are no significant differences. Only two genotypes (SCC 19 and SCA 6) showed pollenkit and only one has perforated walls (SCA 6). Conclusions: The structure and micromorphology of the anthers of T. cacao are similar to those described for other Malvaceae. Likewise, the pollen grains showed variations in size, ornamentation of the sporoderm and the lumen of the reticulum and the presence of pollenkitt. However, no relationship was observed between the micromorphological characters analyzed in the pollen grains and the pollen compatibility models reported for these genotypes.

AISLAMIENTO E IDENTIFICACIÓN DE Lactobacillus spp. (LACTOBACILLACEAE) RESISTENTES A Cd(II) Y As(III) RECUPERADOS DE FERMENTO DE CACAO / Isolation and identification of Lactobacillus spp. (Lactobacillaceae) resistant to Cd(II) and As(III) from fermented cocoa

RESUMEN El objetivo de este estudio fue aislar e identificar a partir de cacao fermentado en Caldas Colombia, bacterias con potencial de aplicación en procesos biotecnológicos, como la detoxificación de cadmio (Cd(II)) y arsénico (As(III)) en el organismo humano. En total se recuperaron 36 aislados de los cuales se recuperaron 11 en presencia de 1,0 mg/L de Cd(II) y 25 en presencia de 0,1 mg/L de As(III). Su identificación molecular determinó que la mayoría de los aislados son del género Lactobacillus. Los ensayos de crecimiento en presencia de diferentes concentraciones de los elementos evaluados permitió determinar que gran parte de los aislamientos presentan resistencia a mayores concentraciones de As(III) (300 mg/L) que de Cd(II) (10 mg/L). En ensayos de tolerancia a la acidez (pH 2,5) se encontró que la cepa tipo Lactobacillus plantarum JCM 1055, junto con los aislamientos nativos L. plantarum A19, A26 y C16, mostraron la mayor tolerancia, por lo que se seleccionaron para evaluar su tolerancia a condiciones de salinidad. Las bacterias evaluadas mostraron crecimiento en concentraciones de hasta 4 g/L de sales biliares. Se concluye que los L. plantarum evaluados en este trabajo tienen un gran potencial para futuros ensayos en los que se busque demostrar la disminución de la bioaccesibilidad de Cd(II) y As(III) en condiciones in vitro del sistema digestivo humano debido a su resistencia a altas concentraciones de estos elementos y su tolerancia a condiciones de acidez y salinidad. Esto, junto con el reconocido potencial probiótico que tienen estos microorganismos, permitirá a futuro su uso en procesos biológicos de mitigación de Cd(II) y As(III).

ABSTRACT The aim of this study was to isolate and identify from fermented cocoa in Caldas Colombia, bacteria with potential application in biotechnological processes such as detoxification of cadmium (Cd(II)) and arsenic (As(III)) in the human organism. In total, 36 isolates were obtained, from which 11 were recovered in the presence of 1.0 mg/L of Cd(II) and 25 in presence of 0.1 mg/L of As(III). Molecular identification showed most isolates belong to the genera Lactobacillus. Minimum inhibitory concentration assays, in presence of different concentrations of the elements, allowed to determine that the majority of isolates have resistance to higher concentration of As(III) (300 mg/L) than Cd(II) (10 mg/L). Acidity tolerance assays at pH 2.5 showed that type strain Lactobacillus plantarum JCM 1055, and native isolates L. plantarum A19, A26, and C16, presented the highest tolerance, thus they were selected to evaluate their tolerance to salinity conditions. The evaluated bacteria could grow in bile salts up to 4 g/L. It is concluded that the evaluated L. plantarum have great potential to be used in assays in which bioaccessibility of Cd(II) and As(III) is diminished under in vitro conditions of the human digestive system, due to its resistance to high concentrations of the elements and tolerance to acidic and high bile salt conditions. These facts, together with the recognized probiotic potential of these microorganisms, may allow their future use in biological processes to mitigate Cd(II) and As(III).

Decrease of the DXR-induced genotoxicity and nongenotoxic effects of Theobroma cacao revealed by micronucleus assay / Diminuição da genotoxicidade induzida por DXR e efeitos não genotóxicos de Theobroma cacao revelados pelo ensaio de micronúcleo

This study evaluated the genotoxicity of lyophilized glycolic extract of Theobroma cacao Linné seeds (TCL), using the micronucleus assay in bone marrow of mice. The interaction between TCL and doxorubicin (DXR) was also analyzed. Experimental groups were evaluated 24-48 h after treatment with N-Nitroso-N-ethylurea (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0.5-2 g/kg), and TCL (2 g/kg) in combination with DXR (antigenotoxic assays). Analysis of micronucleated polychromatic erythrocytes (MNPCEs) showed no significant differences between all the treatment doses of TCL and NaCl control. Mice experimentally treated with DXR and NEU significantly induced MNPCEs. However, a significant reduction of MNPCEs was also observed when TCL was administered in combination with the chemotherapeutic agent DXR. The analysis of the PCE/NCE ratio revealed no significant differences between the NaCl control, all doses of TCL, and DXR. However, there were significant differences in the PCE/NCE ratio between positive NEU control and all other treatments. The PCE/NCE ratio observed after treatment with TCL and DXR showed significant differences and intermediate values to controls (NaCl and NEU). This study suggests absence of genotoxicity and cytotoxicity of TCL, regardless of dose, sex, and time. TCL reduced genotoxic effects induced by DXR, suggesting potential antigenotoxic effects.

Este estudo avaliou a genotoxicidade do extrato glicólico liofilizado de sementes de Theobroma cacao Linné (TCL), usando o ensaio do micronúcleo em medula óssea de camundongos. A interação entre TCL e doxorrubicina (DXR) foi também analisada. Grupos experimentais foram avaliados 24-48 h após tratamento com N-Nitroso-N-etilureia (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0,5-2 g/kg), e TCL (2 g/kg) em combinação com DXR (ensaio antigenotóxico). As análises de eritrócitos policromáticos micronucleados (EPCMNs) não mostraram diferenças significantes entre todas as doses de tratamento do TCL e o controle NaCl. Camundongos experimentalmente tratados com DXR e NEU induziram significativamente EPCMNs. Contudo, uma redução significante de EPCMNs foi também observada quando TCL foi administrada em combinação com o agente quimioterapêutico DXR. As análises da relação EPC/ENC (eritrócito policromático/eritrócito normocromático) revelaram ausência de diferenças significantes entre o controle NaCl, todas as doses de TCL e DXR. Contudo, houve diferenças significantes na relação EPC/ENC entre o controle positivo NEU e todos os outros tratamento. A relação ECP/ENC observada após o tratamento com TCL e DXR mostrou diferenças significantes e valores intermediários aos controles (NaCl e NEU). Este estudo sugere ausência de genotoxicidade e citotoxicidade de TCL, independentemente da dose, sexo e tempo. TCL reduziu os efeitos genotóxicos induzidos por DXR, sugerindo potencial efeitos antigenotóxicos.

Pneuma: receta colonial eficaz en el tratamiento de la disnea / Pneuma: colonial prescription effective in treatment of dyspnea

Objetivo: el archivo histórico Cipriano Rodríguez Santa María de la Universidad de La Sabana cuenta con múltiples recetas médicas empleadas por médicos y boticarios en el Nuevo Reino de Granada. Una de ellas prescribe el cacao para disminuir síntomas generados por afecciones del sistema respiratorio. Objetivos: describir el análisis de una receta médica empleada entre los siglos XVIII y XIX para el tratamiento del ahogo y describir la propiedad fitoterapéutica del cacao conocida gracias a la evidencia científica actual. Materiales y métodos: búsqueda documental en el Archivo Histórico Cipriano Rodríguez Santamaría de la Biblioteca Octavio Arizmendi Posada de la Universidad de La Sabana. Transcripción y análisis del documento denominado "Ahogo" y revisión de la literatura científica actual, así como de los textos originales, sin límite de tiempo. Resultados: la receta médica describe las características del cacao (Theobroma cacao) como agente fitoterapéutico con propiedades antiinflamatorias, analgésicas, antioxidantes e inmunomoduladoras aportadas por componentes como los ácidos oleico, esteárico, palmítico y flavonoides entre otros. La teobromina ejerce efectos broncodilatadores y antitusivosConclusiones: el legado histórico colonial conservado en los archivos locales permite comprender racionalmente las propiedades de los agentes herbarios empleados para enfermedades y síntomas asociados. Existe evidencia científica que respalda el uso del cacao para disminuir la sintomatología asociada con el ahogo. Pese a ello, la escasa o nula descripción de la posología y los efectos secundarios, dificulta analizar la eficacia de esta tradición. En consecuencia, no se puede establecer su eficacia científicamente.

Objetive: The Cipriano Rodríguez Santamaria Historical Archive at Universidad de La Sabana includes multiple medical prescriptions used by physicians and apothecaries in the New Kingdom of Granada. Among them, cocoa was prescribed to relieve symptoms generated by respiratory diseases. Objectives: to describe the analysis of a medical prescription used between the 18th and 19th centuries to treat shortness of breath ("ahogo") and to describe the phytotherapeutic properties of cocoa through current scientific evidence. Materials and Methods: a documentary search in the Cipriano Rodríguez Santamaria Historical Archive: Octavio Arizmendi Posada Library, Universidad de La Sabana. Transcription and analysis of the document named "ahogo" and review of the current scientific literature, as well as, of the original texts, with no time limit. Results: the medical prescription describes the characteristics of cocoa (Theobroma cacao) as a phytotherapeutic agent featuring anti-inflammatory, analgesic, antioxidant and immunomodulatory properties provided by components such as oleic, stearic, palmitic and flavonoid acids, among others. Theobromine exerts a bronchodilator and antitussive effect. Conclusions: the colonial historical legacy preserved in local archives allows a rational understanding of the properties of herbal agents as treatment for diseases and their symptoms. There is scientific evidence supporting the use of cocoa to reduce the symptoms associated with dyspnea. However, little or no description of dosage and side effects makes it difficult to analyze the efficacy of this tradition. Consequently, its efficacy cannot be scientifically established.

Heavy metals content in soils and cocoa tissues in Magdalena department Colombia: emphasis in cadmium / Contenido de metales pesados en suelos y tejidos de cacao en el departamento del Magdalena, Colombia: énfasis en cadmio / Teor de metais pesados em solos e tecidos de cacau no departamento de Magdalena, Colômbia: ênfase no cádmio

ABSTRACT Cocoa (Theobroma cacao L.) is a species originating from the humid tropics. Over 70,000 km2 worldwide is dedicated to growing cocoa, and approximately 30% of its global production is concentrated in the equatorial regions of Central and South America, the Antilles and tropical Asia. The demand for cocoa is growing significantly but the presence of cadmium (Cd) in the species is a potential problem limiting its commercialization. The present study determined the Cd, Ni, Pb and Cr content in two cocoa-producing regions in northern Colombia. Foliage (leaf, bean and shell) and soil were analyzed. The mean Ni, Pb and Cr content in the plant tissue did not exceed the maximum permissible limits set by the United States Environmental Protection Agency (EPA). The mean Cd content in the beans was 0.51 mg kg-1 in region 1 and 0.66 mg kg-1 in region 2, which are considered risky per reports from the European Union and restrict the product's export. These results serve as a reference for future research on cocoa.

RESUMEN El cacao (Theobroma cacao L.) es una especie originaria de los trópicos húmedos. Más de 70,000 km2 en todo el mundo están dedicados a su cultivo y aproximadamente el 30% de su producción mundial se concentra en las regiones ecuatoriales de América Central y del Sur las Antillas y Asia tropical. La demanda de cacao está creciendo significativamente, pero la presencia de cadmio (Cd) en la especie es un problema potencial que limita su comercialización. El presente estudio determinó el contenido de Cd, Ni, Pb y Cr en dos regiones productoras de cacao en el norte de Colombia. Se analizaron el tejido (hoja, frijol y cáscara) y el suelo. El contenido medio de Ni, Pb y Cr en el tejido vegetal no excedió los límites máximos permisibles establecidos por la Agencia de Protección Ambiental (EPA) de los Estados Unidos. El contenido medio de Cd en los frijoles fue de 0,51 mg kg-1 en la región 1 y 0,66 mg kg-1 en la región 2, que se consideran riesgosos según los informes de la Unión Europea y restringen la exportación del producto. Estos resultados sirven como referencia para futuras investigaciones sobre el cacao.

RESUMO O cacau (Theobroma cacao L) é uma espécie originária dos trópicos úmidos. Mais de 70.000 km2 em todo o mundo são dedicados à sua cultura e aproximadamente 30% de sua produção global está concentrada nas regiões equatoriais da América Central e do Sul, Antilhas e Ásia tropical. A demanda por cacau está crescendo significativamente, mais a presença de cádmio (Cd) nas espécies é um problema potencial que limita sua comercialização. O presente estudo determinou o teor de Cd, Ni, Pb e Cr em duas regiões produtoras de cacau no norte da Colômbia. A folhagem (folha, feijão e casca) e o solo foram analisados. O teor médio de Ni, Pb e Cr no tecido vegetal não excedeu os límites máximos permitidos estabelecidos pela Agência de Proteção Ambiental dos Estados Unidos (EPA). O conteúdo médio de Cd nos grãos foi de 0,51 mg kg-1 na região 1 e 0,66 mg kg-1 na região 2, que são considerados de risco por relatórios da União Europeia e restringem a exportação do produto. Esses resultados servem como referência para futuras pesquisas sobre cacau.

Shade and Irrigation Effects on Growth, Flowering, Pod Yields and Cacao Tree Survival Following 5 Years of Continuous Dry Season Irrigation

The effects of plantain shade and dry season irrigation on the growth, field survival, flowering and pod production of cacao was investigated. Treatments were a 2 by 2 factorial combinations of shade regimes (Unshaded/open sun and shaded) and irrigation intervals (5-day and 10-day intervals) arranged in a split-plot design. There was an unirrigated but shaded control. The shade regimes constituted the main plot while irrigation intervals were the sub-plot treatments. The growth, dry season survival, flowering and pod/bean yield characters of cacao were enhanced in the unshaded (open sun) compared with the shaded plants. The open sun treatment combined with 5-day irrigation produced the largest canopy development, flowering and pod production compared with shading-irrigation combinations. The shade-irrigation ameliorated microclimate and enhanced growth and development, flowering and uniform fruiting/pod production and total bean yield and reduced dry season mortality (whole tree death, branch and twig dieback). For the non-irrigated but shaded cacao, about 30% dry season mortality (branch and twig dieback) were obtained. Air temperatures within the cacao field were highest for open sun cacao followed by moderate and dense shade respectively. Flowers were more profuse for unshaded (open sun) cocoa compared with the shaded while the yield and yield components of cacao for each harvest dates and total pod and bean yields were significantly different between the unshaded and shaded cacao regimes. Trees that were irrigated at 5-day intervals produced significantly (P < 0.05) higher LAI, branching, flowers and pods compared with those irrigated at 10-day intervals. The 5-day irrigation interval significantly increased percentage of trees bearing flowers and pods, and produced larger number, and heavier pods and beans compared with the 10-day interval. The drip irrigation strategy adopted ameliorated dry season terminal drought (hydrothermal stresses) in cacao. This is a veritable tool to scale up growth, survival, establishment and flower/pod production.

RP-HPLC-DAD determination of free amino acids in cocoa samples during fermentation and roasting / Determinación de aminoácidos libres en muestras de cacao durante la fermentación y el tostado por cromatografía líquida con detección DAD / Determinação de aminoácidos livres em amostras de cacau durante a fermentação e torrefação por cromatografia líquida com detecção DAD

Abstract Amino acids (AA) composition in cocoa beans can predict the synthesis of compounds which affect cocoa flavor. Thus, their determination is of great interest for the community implied in the commercialization and production of cocoa. In consequence, in this work, the analysis of AA produced during cocoa beans fermentation and roasting was carried out. A high-performance liquid chromatographic method with DAD detection at 254 nm was optimized and validated for their selective determination in six varieties of cocoa beans with different genotypes, all of them grown in Venezuela. AA were extracted by defatted milled cocoa powder ultrasonication using purified water at 70 °C. Then, they were derivatized with phenyl isothiocyanate, and their derivatives were separated, using a reversed-phase column with gradient elution, achieving a satisfactory resolution among the peaks (greater than 1.0) in less than 29 min. 110 cocoa samples were analyzed. Results showed a significant content of free AA, ranging from 3.87 to 5.97 g/kg in absence of fermentation with a predominance of acidic AA. Moreover, there is a progressive increase in the AA content while fermentation process occurs, with a predominance of hydrophobic AA such as alanine, valine, isoleucine, leucine, phenylalanine, and tyrosine. On the other hand, all cocoa types showed a partial degradation of free AA during the roasting step, especially the hydrophobic ones.

Resumen La determinación de aminoácidos (AA) en granos de cacao es de gran interés ya que estos son considerados como unos de los precursores de su sabor y aroma. Por esta razón, el presente trabajo tuvo como objetivo optimizar y validar un método por cromatografía líquida con detección DAD a 254 nm para la determinación selectiva de AA durante la fermentación y tostado en seis variedades de granos de cacao con diferentes genotipos, todos estos cultivados en Venezuela. Los AA se extrajeron del polvo de cacao molido y desgrasado con agua pura a 70 ºC, utilizando la técnica de ultrasonido. Luego, se derivatizaron con fenilotiocianato para separar sus derivados con buena resolución en menos de 29 min en una columna de fase reversa, utilizando gradiente de elución. Se analizaron 110 muestras de cacao. Los resultados mostraron un contenido significativo de AA libres, entre 3,87 y 5,97 g/kg, en ausencia de fermentación con predominio de AA ácidos, y un aumento progresivo en el contenido de AA, mientras ocurre el proceso de fermentación, con un predominio de AA hidrófobos como alanina, valina, isoleucina, leucina, fenilalanina y tirosina. Además, todos los tipos de cacao mostraron una degradación parcial de AA libres durante la etapa de tostado, especialmente los AA hidrófobos.

Resumo A determinação dos aminoácidos (AA) nos grãos de cacau é importante, pois são considerados um dos precursores de seu sabor e aroma. Neste trabalho, um método foi otimizado e validado por cromatografia líquida com detecção DAD a 254 nm para a determinação seletiva de AA durante a fermentação e torrefação em seis variedades de grãos de cacau com diferentes genótipos, todos cultivados na Venezuela. Os AAs foram extraídos do pó de cacau moído e desengordurados com água pura a 70 ºC usando a técnica de ultrassom. Em seguida, foram derivatizados com feniltiocianato, e os derivados foram separados com boa resolução em menos de 29 minutos em uma coluna de fase invertida usando eluição em gradiente. Foram analisadas 110 amostras de cacau. Os resultados mostraram um conteúdo significativo de AA livre entre 3,87 e 5,97 g/kg na ausência de fermentação com predominância de AA ácidos e um aumento progressivo no conteúdo de AA enquanto o processo de fermentação ocorre com predominância de AA hidrófobos como alanina, valina, isoleucina, leucina, fenilalanina e tirosina. Além disso, todos os tipos de cacau apresentaram uma degradação parcial do AA livre durante a fase de torrefação, principalmente o AA hidrofóbico.

Fatty acid profile and solid fat content of Peruvian cacao for optimal production of trade chocolate / Composicion de acidos grasos y contenido de solidos grasos de cacao peruano para la produccion de chocolate óptimo

Using technical procedures, the fatty acid (FA) profile and solid fat content (SFC) of the Peruvian cultivar cacao beans CCN 51 and ICS 6 and the "optimal chocolate", obtained from the mixture of the first two, were determined to assess their quality. These cacao beans were found to have important nutritional values. The FA profile of the cacao beans were similar (p>0.05); however, in the FA profile, the 'optimal chocolate' had significant differences (p≤0.05) in terms of palmitic, arachidic and linolenic acid. The n6:n3 ratio for "optimal chocolate" was 12.0 ± 1.7. Cacao beans had the same SFC, and SFC was highly temperature dependent, as determined using a mathematical model for chocolate. The SFC of chocolate refers to hard cacao butter content at temperatures between 20 and 25°C, and solid fat was heat resistant from 25 to 30°C, which is considered valuable in trade chocolate production. The quality-related properties of these lipid fractions imparted nutritional and physical aspects to the optimal dark chocolate for human consumption.

La composición de ácidos grasos (CAG) y el contenido de sólidos grasos (CSG), de la fracción lipídica de los cultivares peruanos de cacao CCN 51 e ICS 6 así como del "chocolate óptimo", obtenido de las mezclas de las primeras dos, fueron determinados por técnicas analíticas para conocer su calidad. Estas variedades tuvieron valores nutricionales importantes. La CAG de los granos de cacao fueron similares, sin embargo la CAG del "Chocolate óptimo" tuvo diferencias significativas (p<0,05) para los acidos grasos palmitico, araquidico y linoleico. El ratio n6:n3 fue de 12,0 ± 1,7. El CSG de los granos de cacao fueron los mismos y tuvo una fuerte dependencia con la temperatura, también se definió un modelo matematico para el chocolate. El CSG le confiere al chocolate una consistencia dura a temperaturas de 20 a 25°C y resistentes al calor de 25 a 30°C, siendo tales propiedades una ventaja en la comercialización de chocolates. La calidad de estas fracciones lipidicas tuvieron aspectos nutricionales y fisicos en el chocolate oscuro para consumo humano.

Anti-depressant activities of Theobroma cacao Extract on Reserpine-induced Depression in Female Wistar Rats.

Background: Depression is one of the most commontypes of neurological disorder, which is a markedpattern of disturbances in emotional behavior, memoryand hedonic processing. Aim and Objectives: Toinvestigate the role of ethanolic extract of Theobromacacao seed on the prefrontal cortex of female Wistarrats following reserpine-induced depression. Materialand Methods: Thirty-six (36) female Wistar rats wereused for this study. They were divided into six (A - F)groups (n = 6). Group A- control, Group B - 0.2 mg/kgreserpine, Group C - 10 mg/kg fluoxetine, Group D -500 mg/kg Theobroma cacao seed extract, Group E -0.2 mg/kg reserpine + 500mg/kg Theobroma cacaoseed extract, Group F - 0.2 mg/kg reserpine + 10mg/kgfluoxetine. Animals were euthanized via cervicaldislocation after the last day of administration and theprefrontal cortex and hippocampus were excised andfixed in 10% formalin solution for routine histologicalprocessing while the part used for biochemical assaywere homogenized in phosphate buffer beforecentrifugation. Results: Morphological alteration andreduced population of prefrontal cortex andhippocampus neurons, reduced protein synthesis, poorbehavioral patterns, reduced neurotransmission andinduction of oxidative stress in reserpine exposedanimal. Conclusion: Theobroma cacao seed extractwas able to mitigate these aberrations.

First record of the cacao plume moth Michaelophorus nubilus (Felder & Rogenhofer) (Lepidoptera: Pterophoridae) in Mexico

Abstract We report field infestation under natural conditions by the cacao plum moth, Michaelophorus nubilus (Felder & Rogenhofer) on leaves of cacao (Theobroma cacao L.; Malvaceae) for the first time in Southern Chiapas, Mexico. The larvae of M. nubilus damages new "flushing" or light green leaves from buds, from either the main or lateral branches of cacao plants.

Optimization of ultrasound assist extraction of polyphenols in cocoa beans / Optimización del proceso de extracción por ultrasonido de polifenoles en granos de cacao

Background: The determination of polyphenols in cocoa beans allows the establishing of antioxidant properties of great benefit to this product. Objectives: Optimization of the ultrasound-assisted extraction method to determine the content of polyphenols in cocoa beans. Methods: Two experiments design stages were carried out to optimize the ultrasound-assisted extraction process of polyphenols from the cocoa bean. In the first experiment design stage, the adequate extraction solvent was determined; for this purpose, five types of solvents were evaluated through a completely random design unrestricted (CRD). In the second experiment stage, a central composite design 22 + star point (with two central points) was used, which was evaluated using the response surface methodology to determine the influence of the temperature, time, and solute / solvent ratio. Results: The experiment found that acetone: water: acetic acid (70: 29.5: 0.5) mixturee, leads to a greater amount of total extracted phenols measured for the Folin-Ciocalteu method. Analysis of variance (ANOVA) found that six significant effects that influence the response variable (total phenols extracted). The main effects were of the three factors and three of their interactions. Conclusions: After the optimizing said factors, an optimal point was found: 39.3 ° C of temperature, 74.5 minutes, and 22.8 mL of solvent per gram of cocoa sample.

Antecedentes: La determinación de polifenoles en granos de Cacao permite establecer propiedades antioxidantes de gran valor agregado a este producto. Objetivo: Optimización del método de extracción asistida con ultrasonido para determinar el contenido de polifenoles en granos de Cacao. Métodos: Se realizaron dos etapas del diseño experimental para la optimización del proceso de extracción de polifenoles del grano de cacao por ultrasonido. En la primera etapa se determinó el solvente de extracción más adecuado evaluando cinco tipos de solventes, mediante un diseño completamente al azar sin restricciones (DCA). En la segunda etapa experimental, se usó un diseño central compuesto 22 + estrella (con dos puntos centrales), que se evaluó mediante la metodología de superficie de respuesta para determinar la influencia de los factores Temperatura, tiempo y relación soluto/solvente. Resultados: Se determinó que la mezcla acetona: agua: ácido acético (70:29.5:0.5) extrae la mayor cantidad de fenoles totales cuantificados por el método de Folin-Ciocalteu. Mediante la aplicación de un análisis ANOVA, se encontró que seis efectos significativos influyen sobre la variable de respuesta (fenoles totales), estos incluyen los efectos principales de los tres factores y tres de sus interacciones. Conclusiones: Después de la optimización de dichos factores se encontró un punto óptimo que corresponde a 39.3 °C de temperatura, 74.5 minutos y 22.8 mL de solvente por gramo de muestra de cacao.

Association Between Chocolate Consumption and Severity of First Infarction

Abstract Background: Cardiovascular diseases, such as acute myocardial infarction, are the main causes of death in the world. The flavonoids present in chocolate can have benefits for people who have risk factors to the development of cardiovascular diseases and have a coadjuvant effect on known therapies. Objective: To analyze the association between chocolate consumption, severity of coronary lesions, risk factors and severity of the first infarction in patients attended at the Cardiology Institute of Santa Catarina and other hospitals in the State of Santa Catarina. Methods: Subanalysis of the Catarina Heart Study cohort, evaluated 350 patients with first myocardial infarction. We evaluated clinical, echocardiographic, hemodynamic laboratorial variables. We used chi square test to evaluate qualitative variables, t student test in the case of parametric variables and U Mann Whitney test in non-parametric variables. We considered significant p < 0,05. Results: Lower prevalence of hypertension (43.2% % vs. 62.3% p = 0.003), diabetes mellitus (13.5% vs. 25.7%, p = 0.027) and smoking (24.3% vs. 37.7%, p = 0.032) among those who consume chocolate. Higher use of alcohol (40.5% vs. 26.4%, p = 0.018) and drugs (9.5% vs. 3.3%, p = 0.023) among those who consumed chocolate. Among the patients who consumed chocolate, there was a negative correlation between amount consumed and Syntax (r = -0.296, p = 0.019). Conclusion: There was association between chocolate consumption and lower prevalence of hypertension, diabetes and smoking. There was no association between amount of chocolate consumed and post-infarction ventricular function and TIMI frame count. Higher prevalence of alcohol and drug use among those who consume chocolate. Negative correlation between Syntax and the amount of chocolate consumed.