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.

Effect of Yashtimadhu Suppository with Cocoa Butter Base in the Pain Management of Parikartika

Parikartika is one of the common ano-rectal anomalies observed in clinical practise. In Ayurveda Samhitas, this condition is mostly mentioned as an Upadrava of Grahani, Atisara, Arshas and as Vamanavirechanavyapat. ‘Kartanavatvedhana’ is described as predominant symptom of Parikartika which means cutting type of pain in the anus. As per the description of signs and symptoms, Parikartika can be co-related to fissure-in ano in modern contexts. Yashtimadhu is the most widely used drug for Parikartika. Acharya Susruta has mentioned the drug Yashtimadhu in Trividhakarma Adhyaya in post-operative pain management and in treatment for Parikartika in Vamana virechana vyapat. Yashtimadhu in the form of Thaila and Ghrita are commonly used. Here, Yashtimadhuchurna was used to prepare a suppository with cocoa butter as its base for the management of Parikartika. Objectives: The objective of the clinical study is to evaluate any difference in the efficacy of Yashtimadhu suppository in treating Parikartika. Materials and Methods: The study was conducted as an open labelled, randomized, single arm clinical study. 40 Patients who are fulfilling the inclusion, exclusion, diagnostic criteria and complying with the informed consent was selected from OPD and IPD of Sri Jayendra Saraswathi Ayurveda College and Hospital [IEC NO- IEC/ SJSACH/ 09/2021, CTRI NO- CTRI/2021/07/035252]. Case proforma was prepared with all the appropriate details of the patient along with history and physical, rectal examination was done to diagnose Parikartika. Relevant laboratory investigations were performed. Yashtimadhu suppository was given twice per day for about 14 days to 40 patients along with 4 follow-ups once a week for one month after the treatment i.e., 7th, 14th, 21st, 28th day. The subjective parameter was pain and objective parameter were ulceration, sphincter tone. Grading was done according to the specified assessment criteria and was statistically analysed. Results: On completion of the study, statistical analysis was done on the data collected. The intervention, Yashtimadhu suppository showed significant difference in the outcome.

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.

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.

Nigeria's cocoa exports: a gravity model approach / Exportações de cacau da Nigéria: uma abordagem do modelo gravitacional

ABSTRACT: What are the major factors affecting Nigeria's cocoa export flows? In answering this question, the authors suggest a commodity-specific gravity model with three different analytical approaches, (the Heckman Sample Selection Model, the Generalised Least Square, and the Poisson Pseudo Maximum Likelihood), based on a period of 24 years of panel data for Nigeria and it's 36 importing partners to estimate the models. The results showed that GDP, exchange rate policy, WTO, EU, and colonial link are positively associated with the Nigerian cocoa export flows. Further, the negative impact of the GDP per capita, landlocked, distance, AU, and ECOWAS are observed. The need for the expansion of exports to the trading partners, especially the EU members (Netherlands, Germany, France, United Kingdom, Belgium, Spain, etc.), Canada, Malaysia, and the USA is particularly highlighted. These results are important for the formulation of future trade policy that could boost up the Nigerian cocoa exports. This would eventually contribute to the diversification of the Nigerian exports and also enhance the country's foreign earnings.

RESUMO: Quais são os principais fatores que afetam os fluxos de exportação de cacau da Nigéria? Ao responder a esta pergunta, os autores sugerem um modelo de gravidade específica de mercadoria com três abordagens analíticas diferentes (o Modelo de Seleção de Amostras de Heckman, o Mínimo Quadrado Generalizado e a Pseudo Máxima Verossimilhança de Poisson), com base em um período de 24 anos de dados em painel para a Nigéria e seus parceiros importadores para estimar os modelos. Os resultados mostram que o PIB, a política cambial, a OMC, a UE e a ligação colonial estão positivamente associados aos fluxos de exportação de cacau da Nigéria. Além disso, é observado o impacto negativo do PIB per capita, sem litoral, distância, UA e CEDEAO. Destaca-se a necessidade de ampliação das exportações para os parceiros comerciais, especialmente os membros da UE (Holanda, Alemanha, França, Reino Unido, Bélgica, Espanha, etc.), Canadá, Malásia e Estados Unidos. Esses resultados são importantes para a formulação de uma política comercial futura que possa impulsionar as exportações de cacau nigeriano. Isso acabaria por contribuir para a diversificação das exportações nigerianas e também aumentar as receitas externas do país.

Isolation, screening and characterization of bacteria associated with cocoa tree roots for different plant growth promotion (PGP) activities

Aims@#Plant growth promoting rhizobacteria (PGPR) is a group of bacteria that colonise plant roots and enhance plant growth by a diverse range of mechanisms. This study aims to determine the capabilities of PGPR isolated from cocoa tree roots and their efficiency in enhancing plant growth under greenhouse conditions.@*Methodology and results@#Eight samples of healthy cocoa tree roots were collected from different locations in Malaysia. Isolated bacteria were screened based on nitrogen fixation, phosphate and potassium solubilization, and catalase activity. The efficiency of purified PGPR was evaluated from pot experiments of cocoa seedlings under greenhouse conditions. Out of 122 isolates, 18 isolates showed several traits of nitrogen fixation, phosphorus and potassium solubilization and were further screened for other plant growth promoting (PGP) traits like catalase and production of indole acetic acid (IAA). Out of all the PGP trait tests, seven isolates showed the most prominent results for in vitro tests and were further tested in vivo for growth promotion of cocoa seedlings under greenhouse conditions. In the presence of bacterial isolates with 2.5 g of inorganic fertilizer, Leclercia adecarboxylata resulted in increases in plant height, leaf number, root length, stem fresh weight and total fresh and dry weight of cocoa seedlings by 15.68%, 17.14%, 9.48%, 5.67%, 11.84% and 25.12%, respectively.@*Conclusion, significance and impact of study@#Based on the result, L. adecarboxylata incorporated with selected carrier material improve cocoa seedling growth and biomass. This formulation also reduces the production cost of inorganic fertilizer and increase the application and development of biofertilizer.

Desenvolvimento de achocolatados em pó com adição de subprodutos de frutas, processados por spray dryer e com modificador reológico / Development of powdered chocolate added with fruit byproducts, processed by spray dryer and rheological modifier

O achocolatado em pó é um dos derivados do cacau com maior inserção econômica e cultural em diversos países. O objetivo deste estudo foi avaliar a adição de ingredientes diferenciados nesse tipo de produto, como modificadores reológicos e fruta, bem como à alteração no tipo de cacau utilizado, ocasionando mudanças sensoriais e nutricionais positivas ao produto. O fruto sugerido neste estudo foi o cupuaçu (Theoboroma grandiflorum), fruto típico da região Norte, que apresenta excelente qualidade nutricional. Foram desenvolvidas 7 formulações de achocolatado por método convencional após simples mistura (padrão, com cacau alcalino, com cacau orgânico, com polpa de cupuaçu, com amido pré-gelatinizado, com amido pré-gelatinizado + polpa de cupuaçu, com goma guar, com goma guar + polpa de cupuaçu) e 4 formulações processadas por spray dryer após a simples mistura (padrão, com polpa de cupuaçu, com amido pré-gelatinizado, com amido + polpa de cupuaçu). Todas as formulações foram avaliadas quanto à composição nutricional, calorimetria exploratória de varredura (DSC), análises físico-químicas, reológica, quantificação dos compostos fenólicos e avaliação da capacidade antioxidante por métodos in vitro. Em seguida foi realizada análise sensorial com as formulações: padrão, com polpa de cupuaçu e com amido + polpa de cupuaçu. O achocolatado padrão apresentou tempo de mistura de 38 min, o que foi utilizado como parâmetro para as demais formulações. Os achocolatados que continham polpa de cupuaçu apresentaram maior teor proteico (14,5 a 16,3 g/100g) quando comparados com o padrão (13,6 g/100g). Todos os achocolatados apresentaram umidade entre 1,2% e 3,7%, e atividade de água entre 0,13 e 0,57, considerados microbiologicamente estáveis, sendo bom para a vida útil do produto. Os achocolatados obtiveram tempo de molhabilidade entre 07:15 min e 15:06 min; solubilidade de 1,56 IR% a 7,44 IR%; tamanho de partícula variando entre 0,216 mm e 0,347 mm (partículas finas). O uso do spray dryer não teve impacto significativo nas características físicas das formulações, assim como a utilização dos diferentes tipos de cacau não afetou a composição nutricional e qualidade física dos achocolatados. Houve aumento (p< 0,05) para o tempo de molhabilidade e solubilidade do achocolatado com cacau orgânico em comparação com o padrão (13:30 e 9:33 min; 2,64 e 1,56 IR%, respectivamente). A transição vítrea variou entre 35,2 a 35,7 mW enquanto o ponto de carbonização ficou entre 237,4 a 243,6 mW, indicando que a adição dos agentes espessantes e/ou do cupuaçu não interferiu (p<0,05) na análise térmica dos achocolatados. Todos os achocolatados diluídos em leite apresentaram-se como pseudoplásticos, com aumento de viscosidade nas menores temperaturas, conforme esperado. O achocolatado com cacau orgânico apresentou o maior teor de compostos fenólicos (8,27 mg AG g-1) enquanto observou-se redução no conteúdo de fenólicos nos produtos processados por spray dryer. Os achocolatados apresentaram capacidade antioxidante entre 31,76 µMETrolox/g e 75,62 µMETrolox/g, pelo método do DPPH. A adição do cupuaçu levou ao aumento da capacidade de sequestro de radicais DPPH quando comprados com o padrão (p<0,05). Não foi observada diferença significativa pelo método FRAP. A avaliação sensorial obteve aceitação situada na região positiva da escala (5 a 7). Os achocolatados formulados apresentam formulações adequadas a sua comercialização, com agregação de valor nutricional e econômico

The chocolate powdered is a cocoa-derived with greater economic and cultural integration in several countries. The objective of this study was to evaluate the addition of different ingredients in this type of product, such as rheological modifiers and fruit, as well as the change in the type of cocoa used, causing positive sensory and nutritional changes to the product. The fruit suggested in this study was cupuassu (Theoboroma grandiflorum), a typical fruit from the northern region, which has excellent nutritional quality. Seven powdered chocolate formulations were developed by conventional method after simple mixing (standard, with alkaline cocoa, with organic cocoa, with cupuassu pulp, pre-gelatinized starch, pre-gelatinized starch + cupuassu pulp, guar gum, with guar gum + cupuassu pulp) and 4 formulations processed by spray dryer after simple mixing (standard, with cupuassu pulp, pre-gelatinized starch, starch + cupuassu pulp). All formulations were evaluated for nutritional composition, differential scanning calorimetry (DSC), physicochemical, rheological analyzes, quantification of phenolic compounds and antioxidant capacity evaluation by in vitro methods. Then, sensory analysis was performed with the formulations: standard, with cupuassu pulp and starch + cupuassu pulp. The standard powdered chocolate had a mixing time of 38 min, which was used as parameter for the other formulations. The powdered chocolate containing cupuassu pulp had higher protein content (14.5 to 16.3 g / 100g) when compared to the standard (13.6 g / 100g). All powdered chocolate presented humidity between 1.2% and 3.7%, and water activity between 0.13 and 0.57, considered microbiologically stable, wich is good for the shelf life of the product. The powdered chocolate obtained wettability time between 07:15 min and 15:06 min; solubility from 1.56 IR% to 7.44 IR%; particle size ranging from 0.216 mm to 0.347 mm (fine particles). The use of the spray dryer had no significant impact on the physical characteristics of the formulations, as well as the use of different types of cocoa did not affect the nutritional composition and physical quality of the powdered chocolate. There was an increase (p <0.05) for the time of wettability and solubility in chocolate powdered formulated with organic cocoa when compared to the standard (9:33 and 13:30 min; IR 2.64 and 1.56%, respectively). The glass transition ranged from 35.2 to 35.7 mW while the carbonization point ranged from 237.4 to 243.6 mW, indicating that the addition of thickening agents and / or cupuassu did not interfere (p <0.05) in the thermal analysis of powdered chocolate. All powdered chocolate when diluted in milk presented as pseudoplastics, with viscosity increase at lower temperatures, as expected. Chocolate powdered with organic cocoa presented the highest content of phenolic compounds (8.27 mg AG g-1) whereas there was a reduction in phenolic content in products processed by spray dryer. The powdered chocolates presented antioxidant capacity between 31.76 µMETrolox / g and 75.62 µMETrolox / g, by the DPPH method. The addition of cupuassu led to increased ability to sequester DPPH radicals when compared to the standard (p <0.05). No significant difference was observed by the FRAP method. Sensory evaluation was accepted in the positive region of the scale (5 to 7). The formulated powdered chocolates have appropriate formulations for marketing, with added nutritional and economic value

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.

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.

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.

Efeitos dos flavonoides do cacau na prevenção e no tratamento de doenças cardiovasculares: uma revisão de literatura / Flavonoids and cardiovascular diseases effects of cocoa flavonoids on prevention and treatment of cardiovascular diseases: A literature review

O objetivo deste estudo foi revisar na literatura científi ca os efeitos dos fl avonoides do cacau na prevenção e no tratamento de doenças cardiovasculares. Foi realizada uma busca sistematizada nas bases de dados PubMed, SciELO e Trip Data Base, incluindo ensaios clínicos que tiveram intervenção com fl avonoides do cacau, em adultos e idosos, realizados entre os anos de 2008 e 2018. Os descritores em saúde utilizados para a pesquisa foram: "fl avonoids", "cocoa", "dark chocolate" e/ou "cardiovascular diseases". Após análise de doze ensaios clínicos, foi observado melhora nos parâmetros antropométricos e de perfi l glicêmico; redução de colesterol total, lipoproteínas de baixa densidade, triglicerídeos, pressão arterial e de pulso; bem como aumento das lipoproteínas de alta densidade. Além disso, houve diminuição de marcadores infl amatórios, de adesão celular, de estresse oxidativo, do índice de amplifi cação, da velocidade da onda de pulso e da endotelina-1; assim como aumento da dilatação fluxo-mediada, do diâmetro da artéria braquial e do fluxo sanguíneo. Os dados encontrados suportam as alegações de que há efeito benéfico do consumo de flavonoides do cacau sobre a prevenção e o tratamento de doenças cardiovasculares

The aim of this study was to review in the scientific literature the effects of cocoa flavonoids on the prevention and treatment of cardiovascular diseases. A systematic search was carried out in the PubMed, Trip Data Base and SciELO database, including clinical trials covering interventions with cocoa flavonoids, in adults and in the elderly, between the years 2008 and 2018. The health descriptors used for the survey were: "flavonoids", "cocoa", "dark chocolate" and/or "cardiovascular diseases". After reviewing 12 clinical trials, we observed improvement in the anthropometric and glycemic profile parameters; reduction in total cholesterol, low density lipoprotein, triglycerides, blood and pulse pressure; as well as increase in high density lipoprotein. Furthermore, there was a decrease in inflammatory, cell adhesion and oxidative stress markers, augmentation index, pulse wave velocity and endothelin-1; there was also an increase of flow-mediated dilation, brachial artery diameter and blood flow. The data found support the claim that there is a beneficial effect in the consumption of cocoa flavonoids on the prevention and treatment of cardiovascular diseases.

Antioxidant and tyrosinase inhibitory activity of face serum containing cocoa pod husk phytosome (Theobroma cacao L.)

High polyphenol content of cocoa pod extract causing it potential to be developed as antioxidant and tyrosinaseinhibitory agent in cosmetic preparations. Phytosome system known could enhance skin penetration of phytoconstituentlike polyphenol-rich extract. The objectives of this research were to formulate phytosome containing cocoa podextract, develop phytosome complex into face serum preparation, and determine antioxidant and tyrosinase inhibitoryactivities of the extract and the formulated serum. The cocoa pod extract was developed into phytosome system bythin-layer method using soy-phosphatidylcholine. The phytosome then develop into face serum formulation usingViscolam MAC 10 as a gelling agent. The antioxidant activity was conducted by 2,2-diphenyl-1-picrylhydrazyl radicalscavenging assay and tyrosinase inhibitory activity was conducted by colorimetric enzymatic assay. The cocoa podextract has very strong antioxidant activity with inhibitory concentration (IC50) of 17.21 ppm. The extract also hastyrosinase inhibitory activity with IC50 of 199.98 ppm. The phytosome complex containing cocoa pod extract andphosphatidylcholine (1:1) has good entrapment efficiency (90.5%) with an average particle size of 672 nm. Theformulated face serum has good physical characteristic and also has antioxidant and tyrosinase inhibitory activitiesthat equal with the marketed product (Hadalabo ultimate whitening milk, Rohto, Indonesia).

Caracterización molecular de los microorganismos presentes durante el proceso fermentativo de los granos de cacao (Theobroma cacao) / Molecular characterization of microorganisms during cocoa-bean fermentation process (Theobroma cacao)

La fermentación de granos de cacao es un proceso espontáneo de post cosecha muy importante para el desarrollo de aroma y sabor a chocolate el cual involucra un sin número de actividades microbianas complejas. En el presente estudio se identifican los microorganismos presentes en granos de cacao antes, durante y después del proceso de fermentación aplicando dos métodos: el análisis de secuenciamiento de ADN y la espectrometría de masas MALDI TOF/TOF. Dentro del grupo de bacterias y levaduras predominantes identificadas por el primer método se encontro a Lactobacillus plantarum (29%), L. brevis (18%), Bacillus cereus (15%), Pediococcus acidilactici (12%), y Pichia kudriavzevii (100%). Asimismo se caracterizó por huella de masas las secuencias peptídicas más importantes de cada cepa identificada. Por otro lado, aplicando el segundo método, se identificaron 57 especies de microorganismos, siendo el 73.7% especies bacterianas y el 26.3% especies de levaduras. Adicionalmente se detectaron secuencias peptídicas de la proteína vicilina responsable del aroma característico de los granos de cacao fermentados y a la proteína albumina de 21KDa.

Cocoa beans fermentation is a spontaneous process of post-harvest very important for the development of chocolate aroma and flavor, which involves a number of complex microbial activities. In this work, we identify the microorganisms present in cocoa beans before, during and after the fermentation process, applying DNA sequencing analysis and MALDI TOF / TOF mass spectrometry. With the first method, the predominant bacteria and yeast identified were Lactobacillus plantarum (29%), L. brevis (18%), Bacillus cereus (15%), Pediococcus acidilactici (12%), and Pichia kudriavzevii (100%). The most important peptide sequences of each identified strain by mass fingerprint were characterized too. By the second method, 51 species of microorganisms being 73.7% bacterial species and 26.3% yeast species were identified. Additionally peptide sequences responsible Vicilin protein characteristic aroma of the fermented cocoa beans and the albumin protein of 21KDa were detected.

Impact of Local Global Warming on Rainfall and Annual Cocoa Water Requirements in the Regions of Lôh-Djiboua and Gôh in West-central Côte d'Ivoire

Aims: To understand the role of the interannual variability of cumulative rainfall and maximum dry sequences in cocoa production in the Centre-Ouest, one of the cocoa basins in Côte d'Ivoire, in order to propose technical routes more adapted to current rainfall conditions. Study Design: Collection, analysis and processing of daily rainfall data collected by the rain gauges at Divo and Gagnoa stations. Location and Duration of Studies: Divo Cocoa Research Station of the National Center for Agricultural Research, between January 2017 and June 2019. Methodology: The rainfall regime of each locality was determined to assess the impact of rainfall changes on the seasonality of rainfall. The interannual variability of rainfall was studied from the reduced centred rainfall indices. The break years in the time series were detected at both stations from the Khrono Stat software. The interannual cumulative rainfall were analysed for each station and compared to the minimum threshold allowed for cocoa trees. The means of the maximum interannual dry sequences and their probabilities of occurrence were determined using the agrometeorological software called Instat + Version 3.37. Results: The rainfall regime in the area studied (west-central Côte d'Ivoire) has not been modified by the post-rupture rainfall recession as is the case in other parts of the country; it remains a bimodal system characterized by two rainy seasons and two dries during the year. The Divo and Gagnoa regions have been facing a general recession in rainfall since 1966 in Gagnoa and 1972 in Divo. However, the locality of Gagnoa has experienced an increase in rainfall since 2000. Most of the rupture detection tests identified rainfall rupture dates identical to those indicated by the interannual variability highlighted by the rainfall indices. In Gagnoa and Divo, the interannual cumulative rainfalls after the years of rainfall break are reduced compared to those before these rainfall accidents. This situation has led to an increase in the maximum interannual dry sequences in the departments studied. Conclusion: Local climate change has created difficult rainfall conditions after years of rainfall break for cocoa trees as their water needs are increasingly reduced, especially in Divo in Lôh-Djiboua where the downward trend in rainfall has been continuous since 1972. In Gagnoa since the beginning of this century, there has been a new wet period that allows rainfall to adequately meet the cocoa tree's water requirements.

Future Impact of Climate Change on the Yield of Cocoa in Ondo State, Nigeria

This study was carried to determine the trend of cocoa yield and climatic variables and assessment of the impact of climate change on the future yield of cocoa in Ondo State, Nigeria. Annual trend statistics for cocoa yield and climatic variables were analyzed for the state using Mann-Kendall test for trend and Sen’s slope estimates. Downscaled data from six Global Circulation Models (GCMs) were used to examine the impact of climate change on the future yield of cocoa in the study area. The results of trends analysis in Ondo State showed that yield decreased monotonically at the rate of 492.18 tonnes/yr (P<0.05). An increased significant trend was established in annual rainfall trend. While Maximum temperature, minimum temperature, and mean temperature all increased at the rate of 0.02/yr (P<0.001). The ensemble of all the GCMs projected a mid-term future decrease of about 9,334 tonnes/yr by 2050 and a long-term future decrease of 13,504 tonnes/yr of cocoa by 2100. The economic implication of these is that, if the projected change in the yield of cocoa as predicted by the ensemble of all the GCMs should hold for the future, it means that Ondo state may experience a loss of about $22,470,018.22 and $32,308,584.32 by the year 2050 and 2100 respectively according to the present price of the commodity in the world market. Measures are to be taken by the government and farmers to find a way of mitigating the impacts of climate change on the future yield of the cocoa study area. This research should be extended to other cocoa producing areas in Nigeria.

Extracción de compuestos solubles de la cascarilla de cacao con CO2 supercrítico. Caso de metilxantinas y grasa / Extraction of soluble compounds from cocoa shell with supercritical CO2 . A methilxantines and fat case

Resumen La extracción supercrítica es una técnica efectiva para separar diversos compuestos desde matrices naturales, por las propiedades que le confiere a los solventes como el CO2en estado supercrítico. El objetivo del presente trabajo fue la extracción supercrítica de teobromina, cafeína y grasa de la cascarilla de cacao con CO2supercrítico. Se propuso un diseño 22 con 5 puntos centrales, en condiciones estáticas de extracción, teniendo como factores de estudio, presión (2 000 psi a 6 000 psi) y temperatura (318 K a 333 K). Se analizaron los residuos en la cámara de extracción para evaluar los porcentajes de remoción y residual de grasa, cafeína y teobromina, respectivamente. Los resultados mostraron un rendimiento entre 1.72 % a 9.57 %, con una razón de remoción para grasa y cafeína de 61.31 % a 94.54 % y 38.52 % a 78.38 %, respectivamente. La teobromina se retuvo en el polvo de la cascarilla con una razón residual mayor a 90 %. Los efectos de las variables de temperatura y presión para la remoción de grasa fueron predichos a través de un polinomio de primer orden con efectos de interacción, no así para cafeína o teobromina. El método de extracción fue eficiente para remover grasa y cafeína, quedando un residuo en la cámara de extracción considerado como rico en compuestos funcionales.

Abstract Supercritical extraction is an effective technique for separating various compounds from natural matrices due to the properties from solvents, such as CO2, in supercritical state. In the present work, the supercritical extraction of theobromine, caffeine and fat from cocoa shell was carried out with supercritical CO2. A 22 design with 5 central points was proposed, with static operation conditions, having pressure (2 000 psi to 6 000 psi) and extraction temperature (318 K to 333 K) as study factors. The residues were analyzed in the extraction chamber to evaluate the removal and residual percentages of fat, caffeine and theobromine, respectively. The results showed a yield between 1.72 % to 9.57 %, with a removal rate for fat and caffeine of 61.31 % to 94.54 % and 38.52 % to 78.38 %, respectively. The theobromine was retained in the shell powder with a residual ratio greater than 90 %. The effects of temperature and pressure for fat were predicted through a first-order polynomial with interaction effects, but not for caffeine or theobromine. The extraction method was efficient to remove fat and caffeine, leaving a residue powder in the extraction chamber considered as high in functional compounds.

Nematode Parasites of Anurans from Three Cocoa Farms in Ondo State, Nigeria

The research was designed to investigate the parasitic fauna of Anurans from cocoa farms in Ondo state. Amphibians are one of the most threatened groups of vertebrates. Many reasons are attributed to the decline of amphibian species such as global warming, habitat destruction and modification, others include: exploitation, pesticideuse, introduced species, ultraviolet-Bradiation (UV-B), pollution, parasites and diseases. A total of 31 frogs from 4 genera, Hemisus, Ptychadena, Ranaand Xenopus and 7 toads from 1 genus, Sclerophrys were examined. 9 frogs were collected from Oluwateru farm at Iwoye Village; 7 frogs were collected from Folorunso farm at Ako-Igbatoro and 15 frogs from Obodulu farm in Idanre. 5 toads were collected from Oluwateru farm at Iwoye Village and 2 toads from Obodulu farm in Idanre. In all cases collection was done between 20:00hrs and 05:00hrs. Collected specimens were transported in sealed but ventilated containers to the laboratory where identification was done to species level. The frogs were anaesthetized until death in absolute chloroform soaked in cotton wool placed inside kill-jar for 3 minutes in the laboratory.Original researchArticle The gastrointestinal tracts were cut open and the contents of the various sections were put into separate Petri dishes containing normal saline. The skin and the bladder were observed directly under a dissecting microscope for the presence of cysts and monogeneans. The parasites were fixed and preserved in 70% alcohol following standard procedure. Parasites recovered from the gastrointestinal tracts of the anurans include Cosmocerca ornata, Deising, 1861,Cosmocerca cummutata, Diesing, 1851 Paracosmocerca mucronata, Kung and Wu, 1945, Ampliceacum africanum, Taylor, 1924, Gendria liberrei Bain and Philipon, 1969 and Chenospirura asturiHsu, 1957Others were Procamallus brevis Kung, 1948 and Camallanus dimitrovi Durette-Desset and Batcharov, 1974. Some of the parasites are zoonotic while a few others are established parasites of African fishes and water Birds raising probable public health concerns from the findings. Further works aimed at unravelling the biodiversity of hosts and parasites in thelush ecosystem of Ondo state, as well as identification of organisms involved in the life cycle are noted.

Diversity of yeasts during fermentation of cocoa from two sites in the Brazilian Amazon

The purpose of this study was to identify the yeasts involved in spontaneous fermentation of cocoa from the Brazilian Amazon region. The fermentation process was carried out experimentally with cocoa seeds from two sites (Medicilândia and Tucumã), State of Pará, northern Brazil, during a six-day period. Totals of 44 yeasts were isolated from Medicilândia and 29 from Tucumã. Molecular identification was carried out by sequencing the D1/D2 region fragment of the rRNA 26S gene, expanded with universal primers for the NL1GC and LS2 eukaryotes. Pichia manshurica and Saccharomyces cerevisiae were identified in Medicilândia and five yeast species (Pichia fermentans, P. kudriavzevii, P. manshurica, S. cerevisiae and Zygosaccharomyces bailii) were identified in Tucumã. The results showed that P. manshurica and S. cerevisiae may have potential for use as starter cultures in future studies to improve the quality of cocoa seeds fermented in the Brazilian Amazon region. (AU)

A proposta deste estudo foi identificar as leveduras envolvidas na fermentação espontânea de cacau da Amazônia brasileira. A fermentação foi realizada em Medicilândia e Tucumã, Pará, Brasil, durante 6 dias. Em total foram obtidos 44 isolados de leveduras de Medicilândia e 29 de Tucumã. A identificação molecular foi realizada por sequenciamento do fragmento da região D1/D2 do gene rRNA 26S, amplificado com primers universais para eucariotos NL1GC e LS2. Em Medicilândia, foram identificadas Pichia manshurica e Saccharomyces cerevisiae. Em Tucumã foram identificadas cinco espécies (Pichia fermentans, P. kudriavzevii, P. manshurica, S. cerevisiae e Zygosaccharomyces bailii). Os resultados sugerem que P. manshurica e S. cerevisiae podem ter potencial para uso como culturas starter em estudos futuros, para melhorar a qualidade das sementes de cacau fermentadas na Amazônia brasileira.(AU)