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Bacillus thuringiensis is a worldwide known bacterium for its capacity to control insect pests thanks to the action of its parasporal crystal. The objective of this paper deals with the history, in some cases unknown, of the study of Bacillus thuringiensis that led it to be a crucial biological alternative in controlling pest insects. How the mode of action for killing insects was understood, as well as the field tests that were carried out to evaluate its effectiveness and to develop the first commercial products, are reflected in this review that presents and discusses the scientific successes and failures that marked the course of B. thuringiensis.

Lactic acid bacteria (LAB) are currently of great importance given their increasing use in the improvement of human and animal health and nutrition. They exhibit complex nutritional requirements, which is the reason why their production costs are high. Research efforts are being made aimed at evaluating different substrates for their production as well as the production of valuable metabolites from them. The purpose of this paper is to expose the main research and development trends for LAB production for industrial purposes with emphasis on the culture media required for their growth. The web of Science databases as well as the Google Patent Search tool were used in order to gather and analyze the scientific and technical information published in the last twelve years relating to LAB and their culture media. The use of milk, industrial cheese whey, cane molasses, hydrolyzed starches, lignocellulosic materials, organic food waste and bovine blood plasma, among others, have been proposed for Lactobacillus cultivation with the purpose of reducing costs and increasing performance in their production. Research groups and centers have the responsibility of intensifying their efforts to offer highly efficient technological alternatives to the industry that allow the production and application of LAB as a growth factor for the food sector. Also, research in prebiotic ingredients or additives derived from LAB that allow the enhancement of the benefits to the consumer must be continued. In this regard, it is necessary to increase the international visibility of Colombian scientific production in this area.

The articular cartilage is prone to suffer lesions of different etiology, being the articular cartilage lesions of the knee the most common. Although most conventional treatments reduce symptoms they lead to the production of fibrocartilage, which has different characteristics than the hyaline cartilage of the joint. There are few therapeutic approaches that promote the replacement of damaged tissue by functional hyaline cartilage. Among them are the so-called advanced therapies, which use cells and tissue engineering products to promote cartilage regeneration. Most of them are based on scaffolds made of different biomaterials, which seeded or not with endogenous or exogenous cells, can be used as cartilage artificial replacement to improve joint function. This paper reviews some therapeutic approaches focused on the regeneration of articular cartilage of the knee and the biomaterials used to develop scaffolds for cell therapy and tissue engineering of cartilage.

Freemartin syndrome is a intersexuality condition developed in many of the female calfs born from heterosexual multiple calvings (male female). This originates in the fetal development between 30 and 40 days of gestation with transplacental exchange of cells through vascular anastomosis, occurring 60XX/XY chimerism in various tissues, and consequent sterility. In this paper was took blood samples from 106 female calves born from a multiple heterosexual calving. DNA was extracted from leucocytes and searched the SRY gene amplification associated with the Y chromosome by PCR and reading in agarose gel. 90 of 106 samples (84.9%) was amplified the SRY gene, verifying the 60XX/XY chimerism, and 16 samples (15.1%) that did not amplify the gene. Cytogenetic analysis using lymphocytes cell cultures showed the presence of Y chromosome in samples positive for SRY and absence of chimerism in SRY negative samples. The postmortem analysis of female reproductive tracts of SRY positive calves, shows anatomical abnormalities such as clitoris hypertrophia and cervical atresia. Histological examination of gonads of these animals confirmed the presence of ovotestis. This study confirms the usefulness of molecular biology techniques as diagnostic tools of the syndrome, for the use of replacement females in cattle.

Among the most common human diseases with immune system compromise are autoimmune diseases, cancer, and the acquired immunodeficiency syndrome (AIDS). Many of these diseases still have no treatment or their therapies have undesirable side effects. This has aroused a great interest in the search for new natural products with therapeutic potential and scientifically proven effects, showing minimal side effects. Formal clinical and pharmacological investigation in various medicinal fungi of the genus Ganoderma (Ganodermataceae) has shown immunomodulatory effects and tumor growth inhibition in mammals, attributable to the presence of immunomodulatory proteins and other secondary metabolites. To date, six fungal immunomodulatory proteins (FIPs) have been reported in Ganoderma. This paper seeks to advance in the discovery of immunomodulatory proteins present in Ganoderma australe, through mycelium transcriptome 454 Roche® pyrosequencing (RNA-seq) and bioinformatics analyses. The results suggest the presence of gene sequences related to an immunomodulatory protein which has been reported in another fungal species Taiwanofungus camphoratus. The candidate gene sequences found in G. australe exhibit high identity values in their amino acid composition and predicted protein secondary structure with the protein reported for Tai. camphoratus. According to present knowledge about the action mechanisms of these proteins, it is possible to suggest that this is a promising molecule for the treatment and prevention of diseases associated with certain immune deficiencies, cancer, and other diseases with compromised immune systems. Future studies are proposed in order to determine its immunomodulatory potential using in vitro and in vivo assays.

Metabolic Flows Balance in Saccharomyces cerevisiae based on Intracellular Compartmentalization Titulo corto: Metabolic Flows Balance in Saccharomyces cerevisiae Resumen: Una de las técnicas más utilizadas para la predicción de producción de bioproductos y distribución intracelular de flujos de microorganismos es el Análisis de Balance de Flujos - FBA por sus siglas en inglés. El FBA requiere de una función objetivo que represente el objetivo biológico del microorganismo estudiado. En este trabajo se propone un nuevo tipo de funciones objetivo basada en la combinación de objetivos de compartimentos físicos presentes en el microorganismo estudiado. Este tipo de funciones objetivo son examinadas junto con un modelo estequiométrico extraído de la reconstrucción iMM904 del microorganismo S. cerevisiae. Su desempeño se compara con la función objetivo más usada en la literatura, la maximización de biomasa, en condiciones experimentales anaeróbicas en cultivos continuos y aeróbicas en cultivos tipo lote. La función objetivo propuesta en este trabajo mejora las predicciones de crecimiento en un 10% y las predicciones de producción de etanol en un 75% respecto a las obtenidas por la función objetivo de maximización de biomasa, en condiciones anaeróbicas. En condiciones aeróbicas tipo lote la función objetivo propuesta mejora en un 98% las predicciones de crecimiento y en un 70% las predicciones de etanol con respecto a la función objetivo de biomasa. Palabras clave: Análisis de Balance de Flujos, FBA, iMM904, S. cerevisiae, Función Objetivo basada en Compartimentos. Abstract: Flux Balance Analysis - FBA - is one of the most used techniques in prediction of microorganism bioproducts. It requires an objective function that represents a biological objective for the microorganism studied. This paper presents a new kind of objective functions based on individual physical compartment objectives present in the microorganism studied. These kind of functions were tested with a stoichiometric model, extracted from iMM904 reconstruction of S. cerevisiae and its performance is compared with the objective function mainly used in literature, growth maximization, in anaerobic and aerobic batch conditions. The proposed objective function, in the present work, outperforms growth predictions in 10% and ethanol predictions in 75% as compared with the results obtained by maximization of growth objective function, in anaerobic conditions. In aerobic batch conditions, the suggested objective function outperform 98% in growth predictions and 70% in ethanol predictions in comparison with growth maximization. Key words: Flux Balance Analysis, FBA, iMM904 reconstruction, Compartment based Objective Function.

Título en ingles: Evaluation of oxygen transfer in a conventional bioreactor with external aerator Resumen: En el cultivo de microorganismos aerobios se ha observado que los biorreactores convencionales conducen a inconvenientes asociados a la limitación de oxígeno. Para dar solución a esta problemática se diseñó, construyó y evaluó un prototipo de biorreactor con aireador externo. Este sistema permitió incrementar la velocidad de transferencia del oxígeno desde la fase gaseosa a la fase líquida, superando la limitación de oxígeno. Sin embargo, esta nueva configuración carece de modelos empíricos o teóricos para predecir la transferencia de oxígeno a través del cálculo del coeficiente global de transferencia de oxígeno (KLa). En ese sentido, en este trabajo se propuso el uso de análisis dimensional (Teorema Pi de Buckingham), como herramienta para evaluar el efecto sobre el KLa de las diferentes variables involucradas en el proceso, como las de operación, las propiedades de los fluidos y dimensiones geométricas. Luego de desarrollar el análisis propuesto, se encontró una correlación de 0.89 entre los valores generados por el modelo y los datos experimentales disponibles. Se encontró que las variables con mayor incidencia sobre el coeficiente KLa son el flujo de medio de cultivo que circula por el aireador externo denominado flujo de recirculación, la presión en el aireador externo y la velocidad de agitación. Palabras clave: aireador externo, coeficiente volumétrico de transferencia de oxígeno (KLa), Teorema Pi de Buckingham. Abstract: In aerobic microorganism culture, it has been observed that conventional bioreactors lead to disadvantages associated with oxygen limitation. To solve this problem it was designed, built and tested a bioreactor prototype with external aerator. This system allowed increasing the speed of oxygen transfer from gas phase to liquid phase, overcoming the limitation of oxygen. However, this new configuration doesn´t have theoretical or empirical models to predict the oxygen transfer by calculating the global volumetric coefficient of transfer of oxygen (KLa). In that way, this paper proposed the use of dimensional analysis (Buckingham´s Pi Theorem), as a tool to evaluate the effect of various operating variables, fluid properties and geometrical dimensions involved in the process over KLa. After developing the proposed analysis, we found a correlation of 0.89 between the values generated by the model and the available experimental data. It was found that the main variables which affect the coefficient KLa are: the medium flow flowing through the external aerator called recirculation flow, the pressure in the external aerator, and the stirring speed. Key words: external aerator, oxygen transfer coefficient (KLa), Buckingham´s Pi Theorem.

Título en ingles: Incursion of Biotechnology in education: Tendencies and implications Cuando Aristóteles quería hablar de los átomos y el vacío, reproducía partes de la historia del atomismo y se enzarzaba en una discusión con Demócrito, que hacía ya tiempo que descansaba en paz. Cuando el matemático griego quería resolver un problema, la manera natural de proceder era empezar dando una relación de la historia del tema en cuestión, cosa que se consideraba parte integral del problema. Helge Kragh Resumen: Este artículo presenta elementos tenidos en cuenta desde la década de los ochenta, especialmente por países como Reino Unido y Estados Unidos de América, para incluir la educación en biotecnología en las escuelas y así responder a las exigencias sociales y de mercado. Se describen eventos que orientaron la constitución de la educación biotecnológica formal y sirvieron como base para su incorporación en otros contextos. Se hace un acercamiento a los resultados alcanzados hasta este momento, y a las actividades y propuestas establecidas para la educación en biotecnología. Se abordaron algunas de las primeras publicaciones que permitieron la aproximación a los conceptos y contextos que tuvieron lugar para que la biotecnología fuera parte de la educación en varios países del mundo. Se destaca la importancia de configurar la didáctica de la biotecnología y sus implicaciones en la formación de profesores, así como en sus prácticas. Palabras clave: educación en biotecnología, historia, didáctica, formación de profesores. Abstract: This paper presents elements that were taken into account from the 80´s decade, especially in countries such as the United Kingdom and United Stated of America, to include Biotechnology education in the schools and in this way respond to the social and the market demands. Events that directed the formal biotechnological education setting up were described, which were used as a basis for its incorporation in other contexts. An approach to the reached results so far as well as the activities and established proposals for Biotechnological education is made. Some of the first publications that allowed the approach to the concepts and contexts that took place so that Biotechnology can be part of education in various countries around the world were tackled. The importance of configuring the didactics of biotechnology and its implications in Teachers´ training as well as in their practices is highlighted. Key words: biotechnology education, history, didactics, teachers´ training.

Título en ingles: Endospore formation in Clostridium and its interaction with solventogenesis Título corto: Esporulación en Clostridium y su interacción con solventogénesis ResumenLa solventogénesis y la esporulación son mecanismos de las células de Clostridium para resistir ambientes hostiles. Este segundo proceso ha sido estudiado utilizando como modelo lo que sucede con Bacillus, aunque se reconocen diferencias marcadas entre los dos géneros especialmente en el inicio, específicamente en los eventos por medio de los que se da la fosforilación del regulador maestro Spo0A. En la actualidad se ha avalado la teoría que afirma que tres histidin quinasas huérfanas, diferentes a las proteínas Spo0B (fosfotransferasa) y Spo0F en Bacillus, son las encargadas de fosforilar en Clostridium de forma directa a Spo0A, que posteriormente activa la transcripción de diferentes factores sigma relacionados, de forma similar en los dos géneros bacterianos. La proteína Spo0A, perteneciente a la familia de reguladores de respuesta, se comporta como una entidad regulatoria global que tiene injerencia sobre los procesos de formación de esporas y solventes, modulando genes necesarios para que se produzcan acetona y butanol, además de genes de esporulación. El entendimiento de este proceso ha llevado a los investigadores a emplear diferentes técnicas moleculares que permitan incrementar la producción de solventes, así como eliminar la propiedad de las células de producir endosporas. Por tal razón, este escrito presenta un resumen de estas dos redes de expresión de genes, conectadas por el regulador maestro Spo0A.Palabras clave: esporulación, solventogénesis, fosforilación, Clostridium, Spo0A.AbstractSolventogenesis and sporulation are mechanisms used by Clostridium cells to resist hostile environments. Sporulation has been studied using as a model what happens with Bacillus, but marked differences were recognized, particularly in the events that led the phosphorylation of the master controller Spo0A. Currently, a theory that claims that three orphan histidine kinases, different from Spo0B (phosphotransferase) and spo0F proteins in Bacillus, phosphorilate directly Spo0A in Clostridium activating transcription of different sigma factors which are similar in the two bacterial genera, has been supported. Spo0A protein, which belongs to the family of response regulators, behaves as an entity that has global regulatory interference on the processes of spore formation and solvents, modulating genes necessary to produce acetone and butanol. The understanding of this process has led researchers to employ different molecular techniques that increase the production of solvents, and remove the property of the cells to produce endospores. Reason why, this paper presents an updated summary of these two gene expression networks, connected by the master regulator spo0A.Key words: sporulation, solventogenesis, phosphorylation, Clostridium, Spo0A

Título en ingles: Characterization of Aspergillus ficuum hydrolytic enzymes produced in solid state fermentation of cold-pressed canola cake Resumen: En el presente trabajo, se describe la caracterización respecto al pH y temperatura de extractos semipurificados mediante ultrafiltración escalonada de las enzimas fitasa, celulasa y xilanasa de Aspergillus ficuum cepa DSM 932 producidos en fermentación en estado sólido (en adelante SSF) y usando torta de canola como sustrato. La fitasa presentó un pH y una temperatura óptima de 5.0 y 60°C respectivamente, en tanto que la celulasa presentó un pH óptimo de 7.0 con una temperatura óptima de 60°C y el extracto de xilanasa un pH óptimo de 5.4 y una temperatura óptima de 45°C. Palabras clave: caracterización enzimas; fitasa; xilanasa; celulasa. Abstract: This paper, describes the characterization respect to pH and temperature of semi-purified extracts by ultrafiltration step of the enzymes phytase, cellulase and xylanase produced by Aspergillus ficuum DSM 932 strain, in solid state fermentation (SSF) using cold-pressed canola cake as substrate. Phytase showed the optimal pH and temperature 6.0 and 60°C, respectively, while cellulase showed a pH optimal of 7.0 with an optimal temperature of 60°C and xylanase extract an optimal pH of 5.4 and an optimal temperature of 45°C

Título en ingles: Prediction of genome scale of Saccharomyces cerevisiae by flux balance analysis Resumen: El microorganismo Saccharomyces cerevisiae cuenta con gran número de modelos biológicos conocidos como reconstrucciones, las cuales pueden ser a escala genómica. De estas reconstrucciones a escala genómica provienen los modelos matemáticos, también llamados modelos estequiométricos. Una de las técnicas más usadas para estudiar estos modelos es el Análisis de Balance de Flujos (FBA). El proposito del FBA es predecir el crecimiento del microorganismo bajo estudio, y la producción y consumo de componentes como el etanol, CO2 glicerol, sucinato, acetato y piruvato. Para determinar si las predicciones obtenidas mediante FBA son únicas se utiliza la técnica de Análisis de Variabilidad Flujos (FVA). El presente trabajo muestra los resultados de aplicar el FBA a la reconstrucción reciente del microorganismo S. cerevisiae, la denominada iMM904 y los compara con un conjunto de datos experimentales presente en la literatura. Este trabajo también estudia la existencia de múltiples predicciones FBA utilizando la técnica FVA. Los resultados ilustran que es posible predecir el crecimiento del microorganimo S. cerevisiae, con errores entre el 11% y 28%; la producción de CO2, con errores entre el 0.3% y 4.5% y la producción de etanol, con errores entre el 11% y 13%. Palabras clave: analisis de balance de flujos, reconstrucción a escala genómica, iMM904, S. cerevisiae. Abstract: Several biological models, named reconstructions, are used for the study of the S. cerevisiae microorganism. The reconstructions can be genomic scaled. Mathematical models are generated from the reconstructions and they are called stoichiometric models. The flux balance analysis (FBA) is one of the tools used for the analysis of these models. The FBA attempts to predict the evolution of the microorganism and the consumption and production of components like glucose, ethanol, glycerol, succinate, acetate and pyruvate. A Flux variability analysis (FVA) is used to determine the uniqueness of the FBA predictions. This paper shows the results of applying FBA to the iMM904 reconstruction of S. cerevisiae and compares them with experimental data from literature. The results in this paper show that it is possible to predict the evolution with errors between 11% and 28% ; the production of CO2 with errors between 0.3% and 4.5%; and the production of ethanol with errors between 11% and 13%, using FBA for the iMM904 model. Keywords: flux balance analysis, genome scale reconstructions, iMM904, S. cerevisiae

Evaluation of Curcuma longa L. plants obtained by tissue culture in organoponic conditions. Resumen: La cúrcuma es una planta utilizada como condimento y colorante que posee múltiples propiedades medicinales. La evaluación de la estabilidad genética y la respuesta en condiciones de producción constituye una etapa muy importante en los sistemas de propagación de plantas por cultivo de tejidos. En este trabajo se evaluó la respuesta morfoagronómica de plantas de cúrcuma obtenidas por cultivo de tejido y por rizomas en condiciones de organopónico. Los resultados muestran una alta supervivencia (100%), para las plantas provenientes del cultivo in vitro, así como para las obtenidas a partir de los rizomas. Se logró un mayor crecimiento de las plantas propagadas por el cultivo de tejidos durante todo el ciclo de cultivo, sin cambios en las características morfológicas evaluadas. Los rendimientos en las plantas provenientes del cultivo de tejidos fueron superiores en comparación con las plantas propagadas por el método tradicional. Palabras clave: cúrcuma; Biotecnología; micropropagación; cultivo in vitro; evaluación morfológica; evaluación agronómica. Abstract: Turmeric is a plant uses like condiment and coloring. It has multiples medicinal properties. The genetic stability and response in production conditions is an important stage in plant propagation system by tissue culture. In this paper was evaluated the morphoagronomic response of turmeric plants obtained by tissue culture in organoponic conditions. Results show a high survival (100%) for a plant from in vitro culture as well as for the plant obtained by rizomes . A bigger growth of the plants obtained by tissue culture was observed during the whole cultured cycle without changes in the morphologycal characteristic evaluated. The yields of the plants obtained by tissue culture were higher that the plants planted by the traditional method. Keys words: turmeric; Biothecnology; micropropagation; field evaluation; morphologycal evaluation; agronomic evaluation.

Título en inglés: Culture of Chlorella vulgaris using soy waste and applying a magnetic field Cultivo de C. vulgaris con soja y CMChlorella ha ocupado la atención de los biotecnólogos al ser una importante fuente de biomasa para la producción de metabolitos de interés químico farmacéutico e industrial; sin embargo, el manejo de cultivos a gran escala sigue siendo un proceso que necesita economizarse, a partir de alternativas viables. Este trabajo presenta un estudio exploratorio en el que se evaluó la viabilidad del uso del residual de la línea de ablandamiento del grano de soya, como medio de cultivo para la microalga Chlorella vulgaris, obteniéndose excelentes valores de densidad celular máxima (Kmáx: 360 x 106 cél.mL-1). Posteriormente se evalúan los efectos de la aplicación de un campo magnético (CM) de 0.03T, obteniéndose densidades celulares máximas en un menor tiempo de cultivo, lo que puede constituir un fundamento esencial para una nueva metodología de cultivo. La calidad de la biomasa de C. vulgaris se evalúa desde el punto de vista bioquímico, lo que permite determinar la concentración de proteínas, carbohidratos y lípidos en la fase exponencial, cuya acumulación se ve favorecida dependiendo del protocolo de aplicación del CM, mientras que la concentración de lípidos es máxima si el CM es aplicado en la fase estacionaria, variando desde 1.70 ±0.02 hasta 3.48 ±0.03 pg cél; lo que constituiría una ventaja para el manejo de cultivos comerciales de esta microalga que depende de su destino. Palabras clave: imán de neodimio, pigmentos, nitrógeno, biomasa, microalga. ABSTRACT Chlorella has occupied the attention of biotechnologists to be an important source of biomass for the production of metabolites with a pharmaceutical, industrial and chemical interest; however, managing large-scale cultures, remains a process that needs to find alternatives become cheaper and viable. This paper presents an exploratory study that evaluated the feasibility of using the residual of soybeans softening, as a culture medium for the microalgae Chlorella vulgaris, yielding excellent values ​​for maximum cell density (Kmax: 360 x 106 cells.mL-1). Subsequently assessed the effects of applying a magnetic field (MF) of 0.03T, obtaining maximum cell densities in a shorter time, which may constitute an essential base for a new culture methodology. The quality of the C. vulgaris biomass was evaluated from a biochemical point of view, determining protein concentration, carbohydrates and lipids concentration in the exponential phase of growth, whose accumulation is favored, depending on the MF application protocol, while the lipid concentration is maximum if the MF is applied during the stationary phase, with values from 1.70 ±0.02 to 3.48 ±0.03 pg cell. It would be an advantage for the management of commercial cultures of Chlorella depending on the biomass final use. Keywords: neodymium magnet, pigments, nitrogen, biomass, microalgae.

Seed proteins play a very important role in germination. Seeds from a single taxon have a highly stable composition of storage protein. This stability is useful in agronomic, ecological, physiological, taxonomic, molecular and phyto-pathological studies. This paper describes the electrophoretic characterisation in acrylamide gels of seed protein from five pine species (Pinus cembroides, P ayacahuite, P durangensis, P engelmanii, P cooperi and P maximartinezii) from México to determine the importance of their protein profile as biochemical markers in taxonomy. The results suggest that pine reserve protein electrophoretic profiles have chemical attributes having taxonomic importance at subgenus and species level.

In this paper the volumetric oxygen transfer (kLa) and overall heat transfer (Ua) coefficients were determined in flasks which were subjected to rotary shaker action, also provided with a temperature control chamber. Likewise, it was determined the effect over such coefficients of some parameters like surrounding temperature, shaking speed, closure type, liquid volume, capacity and baffles presence or absence in each flask, to determine the optimal work conditions in the rotary shaker. The used liquid in these experiments was distilled water. The kLa and Ua coefficients were also determined in the alcoholic fermentation of glucose by Saccharomyces cerevisiae yeast, in order to establish comparison standards. The maximum and minimum values of referred coefficients to distilled water and the operation conditions were: kLa of 6.2 x 10-3 s-1, working at 25 ºC, 100 rpm, 50 ml of liquid volume, erlenmeyer of 500 ml with baffles and plastic closure. kLa of 4.107 x 10-3 s-1, working at 45 ºC, 60 rpm, 150 ml of liquid volume, erlenmeyer of 250 ml without baffles and cotton closure. Ua of 31.9963 J/min °C, working at 45 ºC, 150 rpm, 150 ml of liquid volume, erlenmeyer of 500 ml with baffles and cotton closure. Ua of 6.0179 J/min °C, working at 35 ºC, 60 rpm, 50 ml of liquid volume, erlenmeyer of 250 ml without baffles and plastic closure. The kLa and Ua values in the alcoholic fermentation and the operation conditions were: kLa of 2.6 x 10-4 s-1 and Ua of 12.8907 J/min °C, working at 35 °C, 150 rpm, 150 ml of liquid volume, Erlenmeyer of 250 ml, with baffles and cotton closure.

In vitro germination of Vanilla planifolia and V. pompona hybridsResumen El cultivo de la vainilla se lleva a cabo ampliamente por medio de propagación clonal, de manera asexual por cortes de tallos y la producción de frutos se realiza por autopolinización. Esta práctica inhibe la variación genética y la emergencia de nuevos individuos por recombinación sexual. Por lo anterior, se considera necesario realizar cruzas por propagación sexual entre especies para obtener nuevos individuos con características deseables para el cultivo. En el presente trabajo se obtuvieron híbridos de dos especies de vainilla como parte de un programa de mejoramiento genético. En general, las semillas híbridas obtenidas que presentaron mayor porcentaje de germinación fueron las cruzas interespecíficas interespecíficos de V. planifolia y V. pompona (85%), seguido de la cruza inversa V. planifolia y V. pompona (57,9%). Las semillas producto de V. pompona autopolinizada obtuvieron valores muy bajos de germinación (10,8%), mientras que las obtenidas de V. planifolia autopolinizada no presentaron germinación. El medio de cultivo más eficiente en todos los tratamientos fue el Murashige & Skoog (MS) adicionado con 400 mg/L-1 de glutamina y 80 mg/L-1 de sulfato de adenina.Palabras clave: vainilla, mejoramiento genético, híbridos de vainilla, germinación in vitro, Vanilla planifolia, Vanilla pompona. AbstractThe cultivation of vanilla is extensive clonal asexual propagation by cuttings and fruit production by artificial self-pollination. This feature tends to inhibit the genetic variation and the emergence of new individuals by sexual recombination. Therefore, sexual propagation between species it is considered necessary to obtain new individuals with desirable characteristics for cultivation. In this paper we were able to obtain hybrids of two species of vanilla. In general, the hybrids seeds obtained whit the higher germination percentage were interspecific crosses of V. planifolia and V. pompona (85%), followed by reverse cross V. planifolia and V. pompona (57.9%). Seeds obtained of V. pompona self-pollination were very low germination (10.8%) while those obtained from V. planifolia self-pollination showed no germination in any media culture. The most efficient medium for all treatments was the Murashige & Skoog (MS) supplemented whit 400 mg/L-1glutamine and 80 mg/L-1 of adenine sulfate. Key words: Vanilla, breeding, vanilla hybrids, in vitro germination, Vanilla planifolia, Vanilla pompona.

Obtaining of cell suspensions and somatic embryos of coffee (Coffea canephora P.) using of bacterial metabolites ResumenLa embriogénesis somática es importante como sistema modelo para estudiar el desarrollo de eventos fisiológicos, citológicos y moleculares que sustentan la embriogénesis en plantas, por ser un sistema adecuado para la propagación masiva de especies vegetales y servir de herramienta para el mejoramiento genético, la conservación de germoplasma y la validación de nuevos productos biológicos, y facilitar la producción a gran escala a través del cultivo en medio líquido y su aplicación en biorreactores, proporcionando alta frecuencia de multiplicación, rápido crecimiento del embrión, facilidad de absorción de nutrientes y reducción de la labor de subcultivo. En este trabajo se empleó la embriogénesis somática como vía de multiplicación para evaluar el efecto de metabolitos bacterianos en la inducción de suspensiones celulares y embriones somáticos en tres genotipos de cafeto pertenecientes a Coffea canephora P. variedad Robusta. Para ello se estudiaron densidades de inóculo entre 0,2, 0,5, 1,0 y 3,0 gMF/L-1, y se evaluó el efecto de diferentes medios de cultivo en el desarrollo del proceso. Los resultados mostraron un comportamiento diferenciado en el genotipo M-28, en medios de cultivo suplementados con reguladores de crecimiento convencionales y en los alternativos. Se evidenció una fuerte relación entre la viabilidad celular y el número de células, ante las diferentes condiciones de cultivo y según la densidad de inóculo, se observó un amplio rango de tamaño y forma en las poblaciones de embriones somáticos. Los porcentajes de conversión de ES con el medio MDE-2 evidenciaron mejoras de este indicador para el cultivo del cafeto.Palabras clave: Coffea canephora, Robusta, densidad de inóculo, germinación, conversión. AbstractThe somatic embryogenesis is important as model system to study the development of physiologic and molecular events that sustain the embryogenesis in plants, is an appropriate system for the massive propagation of vegetable species and as tool for the genetic improvement, the germplasm conservation and the validation of new biological products and to facilitate the multiplication to great scale through the culture in liquid medium, as well as application in bioreactores, providing high multiplication frequency, quick growth of the embryo, easiness of absorption of nutritious and reduction of the subculturing. In this paper the somatic embryogenesis was used to evaluate the effect of bacterial compounds in the induction of cellular suspensions and somatic embryos in three coffee genotypes of Coffea canephora P. var. Robusta. Were studied inoculo densities among 0.2, 0.5, 1.0 and 3.0 gMF/L-1 and the effect of different culture medium in the development of the process. The results showed a behavior differed in the genotype M-28, in medium culture with conventional regulators of growth and the alternatives. Strong relationship was evidenced between the cellular viability and the number of cells, in the different cultivation conditions and according to the inoculo density, a wide range of size and forms as observed in the populations of somatic embryos. The conversion percentages with the medium MDE-2, evidenced improvements of this indicator for the coffee. Key words: Coffea canephora, Robusta, density, germination, conversion.

Título en inglés: Culture of Chlorella vulgaris using soy waste and applying a magnetic field Cultivo de C. vulgaris con soja y CM Chlorella ha ocupado la atención de los biotecnólogos al ser una importante fuente de biomasa para la producción de metabolitos de interés químico farmacéutico e industrial; sin embargo, el manejo de cultivos a gran escala sigue siendo un proceso que necesita economizarse, a partir de alternativas viables. Este trabajo presenta un estudio exploratorio en el que se evaluó la viabilidad del uso del residual de la línea de ablandamiento del grano de soya, como medio de cultivo para la microalga Chlorella vulgaris, obteniéndose excelentes valores de densidad celular máxima (Kmáx: 360 x 106 cél.mL-1). Posteriormente se evalúan los efectos de la aplicación de un campo magnético (CM) de 0.03T, obteniéndose densidades celulares máximas en un menor tiempo de cultivo, lo que puede constituir un fundamento esencial para una nueva metodología de cultivo. La calidad de la biomasa de C. vulgaris se evalúa desde el punto de vista bioquímico, lo que permite determinar la concentración de proteínas, carbohidratos y lípidos en la fase exponencial, cuya acumulación se ve favorecida dependiendo del protocolo de aplicación del CM, mientras que la concentración de lípidos es máxima si el CM es aplicado en la fase estacionaria, variando desde 1.70 ±0.02 hasta 3.48 ±0.03 pg cél; lo que constituiría una ventaja para el manejo de cultivos comerciales de esta microalga que depende de su destino. Palabras clave: imán de neodimio, pigmentos, nitrógeno, biomasa, microalga. ABSTRACT Chlorella has occupied the attention of biotechnologists to be an important source of biomass for the production of metabolites with a pharmaceutical, industrial and chemical interest; however, managing large-scale cultures, remains a process that needs to find alternatives become cheaper and viable. This paper presents an exploratory study that evaluated the feasibility of using the residual of soybeans softening, as a culture medium for the microalgae Chlorella vulgaris, yielding excellent values ​​for maximum cell density (Kmax: 360 x 106 cells.mL-1). Subsequently assessed the effects of applying a magnetic field (MF) of 0.03T, obtaining maximum cell densities in a shorter time, which may constitute an essential base for a new culture methodology. The quality of the C. vulgaris biomass was evaluated from a biochemical point of view, determining protein concentration, carbohydrates and lipids concentration in the exponential phase of growth, whose accumulation is favored, depending on the MF application protocol, while the lipid concentration is maximum if the MF is applied during the stationary phase, with values from 1.70 ±0.02 to 3.48 ±0.03 pg cell. It would be an advantage for the management of commercial cultures of Chlorella depending on the biomass final use. Keywords: neodymium magnet, pigments, nitrogen, biomass, microalgae.

Título en inglés: Evaluating oxygen transfer in a Lactococcus lactis cultures using an external aeration fermentation system (EAFS) Resumen En fermentaciones aerobias el oxígeno, como aceptor terminal de electrones en el proceso de respiración, comúnmente se constituye en limitante debido entre otros factores al diseño del biorreactor (factores geométricos), a las condiciones de operación de los fermentadores (condiciones ambientales requeridas en el cultivo, potencia transferida al cultivo por el sistema de agitación, propiedades del medio líquido), demanda de oxígeno por parte del microorganismo, sistema de aireación (concentración de oxígeno en el gas, solubilidad del oxígeno). La limitación de oxígeno se refleja en la fermentación con Lactococcus lactis cepa IBUN 34.1, en que presenta una baja disponibilidad de oxígeno desde muy temprano en la fase exponencial del cultivo. Para superar estas limitaciones se diseñó y desarrolló un sistema de suministro de oxígeno de alta tasa de transferencia, consistente en un sistema de fermentación con aireación externa (SFAE), el cual es comparado en este trabajo con el sistema tradicional de fermentador agitado dotado con dos turbinas tipo Rushton y aireación por difusor interno. En este trabajo se evalúa la operación del SFAE, se seleccionan y estudian algunas variables operacionales y su efecto sobre la transferencia de oxígeno gas-líquido. Los resultados indican que las variables que tienen efecto significativo sobre el coeficiente volumétrico global de transferencia de masa kLa son la agitación y el flujo de medio de cultivo que circula por el aireador externo denominado flujo de recirculación. Los valores de kLa obtenidos indican que con el fermentador convencional con aireación interna el mayor valor de kLa alcanzado fue de 40,68 (h-1), en tanto que con el SFAE se alcanzaron valores de 63,18 (h-1). Palabras clave: biorreactores; kLa; transferencia de oxígeno; fermentaciones aeróbicas; transferencia de masa gas-líquido. Abstract In aerobic fermentations, oxygen as terminal electron acceptor in respiration process, is commonly a limiting due to factors like its low solubility in aqueous solutions, bioreactors’ geometric factors and operating characteristics, liquid media properties, oxygen concentration in gas supply, microorganisms’ characteristics, environmental culture conditions, power supply by agitation system, etc. Oxygen limitation is present in cultures using the IBUN 34.1 Lactococcus lactis strain where oxygen availability is low some minutes after the exponential phase starts. A high transfer rate oxygen supply system was thus designed to overcome such limitations; it consisted of an external aeration fermentation system (EAFS) which was compared in this work with a conventional agitated tank fermenter equipped with two Rushton turbines and internal diffuser aeration flute mechanism. This paper evaluates the EAFS; some operational variables were selected and studied as well as their effect on oxygen transfer. Our results showed that agitation and culture medium flow through the aerator (called external recirculation flow) were the variables having the main effect on overall volumetric mass transfer coefficient (kLa). The highest kLa value in the conventional bioreactor having internal aeration was 40.68 kLa (h-1), while the EAFS reached 90 (h-1). Key words: Bioreactor; kLa; oxygen transfer; aerobic fermentation; gas–liquid mass transfer.

Expresión transitoria GUS en callos de arroz (var. J-104) mediante la optimización de las condiciones de cultivo in vitro Resumen This paper purposes suitable conditions for callus induction and co-cultivation with Agrobacterium tumefaciens of J-104 rice cultivar. It was evaluated the effect of different concentrations of 2.4-D and agar, and the inclusion of L-proline and L-glutamine in callus culture medium. The use of 2.5 mg/L 2.4-D and 0.8% agar allowed the highest percentage of embryogenic calli. Callus formation was improved considerably with 500 mg/L of L-proline and L-glutamine in the culture medium. Different factors were studied throughout co-cultivation of calli with A. tumefaciens: inoculation time, co-cultivation temperature, concentration of acetosyringone and co-cultivation period. Transient GUS expression was quantified by fluorometry in all co-cultivated calli. The best results were obtained with the following conditions: 10 min as inoculation time, 100µM acetosyringone in co-cultivation medium, temperature of 20ºC, and 3 days as co-cultivation period. Key words: Agar; callus; co-cultivation; fluorometric GUS activity. Resumen Se describen las condiciones óptimas para la callogénesis y cocultivo de callos con Agrobacterium tume-faciens de la variedad de arroz J-104. Se determinó el efecto de diferentes concentraciones de 2.4-D, agar y de L-prolina y L-glutamina en el medio de cultivo de callos. El uso de 2,5 mg/L de 2.4-D y 0,8% de agar permitió lograr el porcentaje más alto de callos embriogénicos. La formación de callos fue mejorada considerablemente con la adición de 500 mg/L de L-prolina e igual concentración de L-glutamina en el medio de cultivo. Se estudiaron diferentes factores en el cocultivo de los callos con A. tumefaciens: tiempo de inoculación, concentración de acetosiringona, temperatura y tiempo de cocultivo. Para comparar el efecto de cada factor sobre la expresión GUS se cuantificó la actividad transitoria mediante fluorimetría. Los valores más altos de actividad fluorimétrica fueron obtenidos con las siguientes condiciones: 10 min de inoculación, 100µM de acetosiringona en el medio de cocultivo y 3 días de cocultivo a 20 ºC. Palabras clave: agar; callo; cocultivo; actividad fluorimétrica GUS.

Excessive application of chemical nitrogen fertilisers and pesticides has badly affected the environment. This has led to great interest being shown in studying a crop's native microbial community and its benefit for plants. This paper was thus aimed at characterising sugarcane's endophytic microbial community. 5 sugar cane strains and 50 isolates were used. Gas chromatography was used for measuring nitrogenase activity and the influence of carbon and nitrogen sources and pH on cultures. Indol acetic (IAA) production was detected by Dot-Immunobinding and Salkowski's method. These results show that 19 strains and isolates had nitrogenase activity, values ranging from 100 to SOOO/zg/mL; 6 of them produced IAA (values ranging from 1,7 to 2,5 //g/mL): Gluconacetobacter diazotrophicus PAl-5, Gluconacetobacter diazotrophicus 1-05, Gluconacetobacter diazotrophicus 4-02,17,30 and 305. It was demonstrated that culture medium nutrient sources and pH affected the nitrogenase activity of the strains representing the endophytic community. Key words: endophytic community, sugarcane, nitrogenase activity, indolacetic acid.

The Meliacea are of great importance in construction and furniture-making. Toona ciliata is a Meliacea originally from the Himalayan region; it is known as Himalayan Cedar in Cuba. Natural regeneration occurs in this specie by seed diffusion and grafting; such propagation is limited. The object of this paper was to promote callus formation and plant regeneration in Toona ciliata from in vitro propagation. Two to three year old mature trees (Toona ciliata) were used. Rachis were taken from young branches from these plants. They were disinfected in 0.25% (w/v) mercuric chloride solution for 10 min followed by three rinses in autoclaved distilled-water. They were then established in MS supplemented with 0-1 mg/L thidiazuron culture medium. Nodular calluses were obtained having good morphogenic characteristics. Shoots sprouted from six-month-old calluses in the dark and plant regeneration was done in the light. AIB shoots were rooted in MS medium supplement with 1 mg/L IBA. Key words: callus, Himalayan Cedar, Meliacea, growth regulators, plant regeneration. Abbreviations: IBA- indolebutyric acid, thidiazuron-N-1,2,3-thiadiazol-5-yl-N-phenylurea.

The Meliacea are of great importance in construction and furniture-making. Toona ciliata is a Meliacea originally from the Himalayan region; it is known as Himalayan Cedar in Cuba. Natural regeneration occurs in this specie by seed diffusion and grafting; such propagation is limited. The object of this paper was to promote callus formation and plant regeneration in Toona ciliata from in vitro propagation. Two to three year old mature trees (Toona ciliata) were used. Rachis were taken from young branches from these plants. They were disinfected in 0.25% (w/v) mercuric chloride solution for 10 min followed by three rinses in autoclaved distilled-water. They were then established in MS supplemented with 0-1 mg/L thidiazuron culture medium. Nodular calluses were obtained having good morphogenic characteristics. Shoots sprouted from six-month-old calluses in the dark and plant regeneration was done in the light. AIB shoots were rooted in MS medium supplement with 1 mg/L IBA. Key words: callus, Himalayan Cedar, Meliacea, growth regulators, plant regeneration. Abbreviations: IBA- indolebutyric acid, thidiazuron-N-1,2,3-thiadiazol-5-yl-N-phenylurea.