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    Seminal plasma composition from ejaculates collected by artificial vagina and electroejaculation in Guirra ram

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    This study was conducted to evaluate changes in ram seminal plasma composition from ejaculates obtained using artificial vagina (AV) and electroejaculation (EE). To address this question, we assessed the effect of semen collection method on volume, sperm concentration, sodium concentration, potassium concentration, sodium/potassium ratio, total protein content and protein profile using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and 2-D polyacrylamide gel electrophoresis. The main findings from this study were: (i) similar volume was obtained, while sperm concentration was significantly lower for EE method; (ii) potassium and sodium/potassium concentration ratio were not influenced by recovery method, while sodium concentration increased significantly when semen was recovered using EE; (iii) approximately 80% of the total relative seminal plasma protein is represented by four protein fractions of molecular weights around 15, 21, 24 and 50 kDa and there were not differences and (iv) focussing the two-dimensional SDS-PAGE gel on the 10-25 kDa rank, the IMAGE ANALYSIS software detected around 22 spots with isoelectric points ranging from 5.1 to 6.1. Two protein spots (15 kDa and 5.5 and 22 kDa and 5.2 for molecular weight and isoelectric point respectively) increased significantly when semen was recovered using EE. One spot protein with molecular weight around 25 kDa and isoelectric point of 5.2 were only found in the seminal plasma from the semen recovery by AV. As it was demonstrated, ejaculates obtained with EE modify the sodium concentration, alter two proteins concentration and induced the loss of one protein in seminal plasma

    Algoritmos de cálculo de las emisiones de metano y óxido nitroso de la gestión de los estiércoles y purines en rumiantes en condiciones españolas

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    La cría intensiva de rumiantes representa una fuente importante de gases de efecto invernadero a la atmósfera. La fermentación entérica es una de las principales fuentes de metano de esta actividad. No obstante, la gestión de las deyecciones ganaderas sólidas (estiércoles) y líquidas (purines) genera también emisiones tanto de metano (CH4) como de óxido nitroso (N2O). Dentro de las emisiones de gases procedentes de la gestión de los estiércoles y purines, los rumiantes se sitúan en segundo lugar de importancia después de la ganadería porcina. El origen de estos gases se encuentra en la descomposición de la materia orgánica que se produce a lo largo de todas las fases de la gestión de las deyecciones ganaderas. La estimación de estas emisiones es obligatoria y debe reflejarse en los Inventarios Nacionales de Emisiones a la Atmósfera. Para ello, el Panel Intergubernamental sobre Cambio Climático (IPCC) proporciona unas ecuaciones de cálculo de las emisiones que dependen de una serie de factores, de los cuales el sistema de gestión del estiércol o purín es determinante. Sin embargo, en la mayoría de los casos, estas ecuaciones no pueden aplicarse a las condiciones españolas debido a la falta de concordancia entre los sistemas de gestión de las deyecciones ganaderas del IPCC y los que se dan actualmente en España. Por ejemplo, en el caso del ganado ovino y caprino, el sistema más habitual (cama profunda) no se contempla, y en el ganado bovino predominan unos sistemas de operaciones secuenciales y concatenadas que no se ajustan a estas ecuaciones tal y como se plantean. El objetivo de este trabajo es presentar unos algoritmos alternativos, para la estimación de las emisiones de CH4 y N2O derivadas de la gestión de los estiércoles y purines de los rumiantes en España

    Citrus tristeza virus resistance in a core collection of sour orange based on a diversity study of three germplasm collections using QTL-linked markers

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    Seven markers linked to QTL involved in CTV accumulation, leafminer resistance and apomictic reproduction were used to characterize 64 sour orange (Citrus aurantium L.) accessions from three national collections in order to identify a representative core in which the resistance behaviour against two Citrus tristeza virus (CTV) isolates was studied. Different degrees of apomixis were found between the foreign collections. Most of the C. aurantium accessions fall into three main groups based on three multilocus genotypes. The haplotype diversity at three CTV accumulation QTL-linked markers was further studied by sequence analysis of alleles. Genotypic and allelic diversity at one of them, tightly linked to Ctv-R(2) in Poncirus trifoliata (L.) Raf., match the plant-CTV interaction types reported among Poncirus and Citrus species. Only those selected accessions from the major group presented CTV resistance during 30 months of continuous growth, but later the resistance broke down in some plants. CTV tolerance appears related to slow growing genotypes. Certain micronutrients: Mn, and B depending on the accession, might play a relevant role in this host-pathogen interaction particularly in alkaline soils

    Automatic inspection of the pomegranate (Punica granatum L.) arils quality by means of computer vision. Short communication.

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    With the aim of ensuring the quality and homogenising the colour of commercial batches of pomegranate arils, a new prototype for the automatic inspection of pomegranate arils and their separation into categories by means of air ejectors has been developed. Arils are singulated and transported on conveyor belts. Images of the arils are acquired using two cameras to allow estimation of the quality and are then expelled through different outlets according to the category. New control software were developed that synchronise the advance of the conveyor belts, the acquisition of images and the separation by air ejectors, which also includes mechanisms for the synchronisation and communication between the vision and control computers. A new computer vision system has been developed for the inspection of arils that distinguish between arils and raw material by estimating the colour of the objects and then classify the arils by size and colour. The prototype was tested over a six-month period in a Spanish pomegranate producer, with the inspection of about 5 Mg of product. The new prototype achieves the objective of inspecting and sorting pomegranate arils according to their quality, reaching the performance specifications

    Effect of two nutrient solution temperatures on nitrate uptake, nitrate reductase activity, NH4(+) concentration and chlorophyll a fluorescence in rose plants

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    The effect of two nutrient solution temperatures (cold (10 degrees C) and warm (22 degrees C)) during two flowering events of rose plants (Rosa x hybrida cv. Grand Gala) were examined by measuring chlorophyll (Ch1) a fluorescence, ammonium (NH4(+)) content and nitrate reductase (NR) activity in four different leaf types, that is, external and internal leaves of bent shoots and lower and upper leaves of flowering stems. Besides, nitrate (NO3-) uptake and water absorption, total nitrogen (N) concentration in the plant, dry biomass, and the ratios of shoot/root and thin-white roots/suberized-brown roots were determined. Generally, cold solution increased NO3- uptake and thin-white roots production but decreased water uptake, so plants grown at cold solution had to improve their NO3- uptake mechanisms to obtain a higher amount of nutrient with less water absorption than plants grown at warm solution. The higher NO3- uptake can be related to an increase in NR activity, NH4(+) content and total N concentration at cold solution. Nutrient Solution temperature also had an effect on the photosynthetic apparatus. In general terms, the effective quantum yield (phi(PSII)) and the fraction of open PSII reaction centres (q(L)) were higher in rose plants grown at cold solution. These effects can be associated to a higher NO3- uptake and total N concentration in the plants and were modulated by irradiance throughout all the experiment. Plants could adapt to cold solution by enhancing their metabolism without a decrease in total dry biomass. Nevertheless. the effect of nutrient solution temperature is not simple and also affected by climatic factors

    Mejora de la eficacia de los tratamientos de desinfección del suelo con 1,3- dicloropropeno y cloropicrina, en condiciones de bajas temperaturas, para cultivos de pimiento y plantas madre de fresón.

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    Algunos cultivos se adaptan mal a la desinfección en invierno debido a que las bajas temperaturas reducen la capacidad de evaporación de los fumigantes con lo que la distribución espacial de los vapores no se puede realizar con la uniformidad deseada. Por otra parte la eliminación de los residuos después de la desinfección se realiza más lentamente y se alarga el tiempo de espera antes de la plantación para evitar problemas de fitotoxicidad. Hay dos casos más problemáticos en la agricultura española: el cultivo de pimiento italiano y el de planta madre de fresón en viveros de altura, que se inician en primavera y en los que la desinfección previa a la plantación se produce con temperaturas del suelo que rayan los 10 ºC. Otro problema lo representa la aparición de malas hierbas en primavera y verano con la plantas madre de fresón aún jóvenes y la aparición de Cyperus rotundus en el cultivo del pimiento. Esta investigación propone el estudio de tres formulaciones de 1,3-dicloropropeno 55.2% + cloropicrina 30.3% a 50 g/m2 facilitados por la compañía Agroquímicos de Levante SA, con la adición o no de un herbicida (trifluralina 48% pv ) al 1% y se comparan con un formulado de dicloropropeno y cloropicrina ya registrado en España y actualmente en uso a base de 1,3-dicloropropeno 55.4% + cloropicrina 32.7% (Agrocelhone NE) a 50 g/m2, bromuro de metilo 50% + cloropicrina 50% a 40 g/m2 y un testigo no desinfectado. Se controló la temperatura del suelo durante la aplicación y hasta 1 semana después. La aplicación en viveros de altura se realizó mediante inyección al suelo y cubierta de PE a máquina, a la manera habitual, en dos pasadas, 20 días después se inició un cultivo de planta madre cv. Elsanta. En el cultivo de pimiento se aplicó mediante inyección a través del riego localizado, bajo el acolchado negro con los lomos dispuestos para el cultivo. 20 días después se procedió al transplante del pimiento cv. Estilo Los resultados indican que algunas de las nuevas formulaciones aumentan la eficacia herbicida hasta niveles comparables al bromuro de metilo. La mezcla registrada de dicloropropeno y cloropicrina se mantiene en un lugar intermedio, con diferencias significativas con el BM, en el caso de viveros de altura, aunque sin diferencias en el cultivo de pimiento. Las nuevas formulaciones sin herbicida, objeto de estudio, no muestran diferencias significativas con el BM, (en viveros de altura) ni desde el punto de vista de la producción ni en el del control de malas hierbas en aplicaciones de invierno, en condiciones de baja temperatura (10 ºC aprox.). Podemos concluir que a la vista de los resultados de este primer año de estudio, algunos coadyuvantes pueden hacer que las mezclas de 1,3-dicloropropeno y cloropicrina mejoren las características de las formulaciones actualmente comercializadas como alternativas al bromuro de metilo

    Las formulaciones emulsionables de Dimetildisulfuro (DMDS) y sus mezclas con cloropicrina como alternativas al bromuro de metilo

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    Desde hace mucho tiempo se sabe que plantas de los géneros Allium sp. y Brassica sp. poseen una cierta capacidad antimicrobiana. Este poder ha sido explicado por la presencia de radicales sulfuro de alilo y sulfuro de metilo derivados de sulfóxido de LCisteina existente en los ajos y las brasicas. Una de las sustancias más interesantes es el Disulfuro de dimetilo (DMDS) que es un importante compuesto de los Allium sp., especialmente del ajo. Aunque sus propiedades fumigantes se conocen desde hace poco tiempo, el DMDS se viene utilizando desde hace mucho en la industria petroquímica e incluso como agente de aromas por su fuerte olor a ajo. El DMDS es un fumigante de amplio espectro con efectos nematicida, fungicida insecticida y herbicida, que ejerce su acción a nivel de respiración mitocondrial bloqueando la actividad citocromo oxidasa de las células. Los efectos nematicidas se han podido comprobar contra Heterodera sp. y Meloidogyne sp., mediante aplicación en preplantación. Se le reconoce eficacia contra hongos del suelo como Sclerotium rolfsii, Rhizoctonia solani, Phytophthora cactorum pero no tan efectivo contra Macrophomina phaseolina y solo parcialmente contra Sclerotinia scleritiorum. Desde el año 2003 al 2007 hemos venido estudiando sus efectos como alternativa al bromuro de metilo en el cultivo de pimiento en las zonas del Bajo Segura (SURINVER, Pilar de la Horadada, Alicante) y en La Canal de Navarrès (VALSUR, SAT Bolbaite, Valencia) usando formulaciones emulsionables aplicadas con el riego localizado, tanto sólo como mezclado con diversas proporciones de cloropicrina (50%; 33%), con objeto de mejorar su eficacia fungicida. El DMDS sólo, a dosis elevadas (105 g/m2), tiene una capacidad herbicida notable; a menor dosis, sus mezclas con cloropicrina poseen un excelente efecto fumigante que mejora cuando se aplica bajo plástico VIF. El efecto fungicida también mejora ostensiblemente. Una vez depurada la técnica de aplicación la calidad y producción de pimiento es comparable a la obtenida con los tratamientos tradicionales con bromuro de metilo

    ACTA HORTICULTURAE

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    Alternatives to synthetic fungicides are needed worldwide to control citrus postharvest diseases. Curing of citrus fruit at 30-37°C and 90-98% RH for 65-72h has been repeatedly reported as an effective method to control green mold caused by Penicillium digitatum. However, commercial adoption of this practice is limited because of the length of treatment and risks of negative impact on fruit quality. Exposure to CO2-enriched atmospheres is known to provide fungistatic effects and, in some cases, to induce resistance in fresh fruit against major postharvest pathogens. To improve curing treatments against citrus postharvest green mold, work is in progress to determine if short treatments with CO2 at curing temperature can control established infections and/or induce fruit resistance. Mandarins ‘Nadorcott’ and ‘Ortanique’ were artificially inoculated with P. digitatum and 24h later, were exposed to air (control) or 15, 30 and 50 kPa CO2 at 20 or 33°C for 8 or 24h. Number of decayed fruit was counted after 4 and 7 d incubation at 20°C. To assess potential induction of resistance and time of induction, the same treatments were applied to another set of non-inoculated fruit. Treated fruit were kept at 20°C for 1, 2 or 5 d, at which time mandarins were wound inoculated with the pathogen. Lesion diameter was then measured after 3 and 6 d at 20°C. For both cultivars, only treatments at 33°C for 24h reduced the incidence of disease significantly after 4 d incubation, with 15 kPa CO2 slightly superior to other gas concentrations used. Regardless of time between treatment and inoculation and incubation period, all treatments were ineffective in reducing disease severity and, therefore, in inducing any kind of fruit resistance. It is planned to evaluate longer exposure times (maximum of 48h) to 15 kPa CO2 at 33°C

    A new Aphytis species on 'Aonidiella aurantti'?

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    In the last surveys of parasitoids conducted on Californian red scale, Aonidiella aurantii (Maskell), individuals of the genus Aphytis that present a black pigmentation of the pupa and exuvia have been detected. This description does not correspond to any of the parasitoids cited in Spain. Aphytis chrysomphali (Mercet), Aphytis melinus DeBach and Aphytis lingnanensis Compere, show a clearly differentiated pupa and exuviae pigmentation, without black pigmentation of pupa head in all cases. Therefore, these individuals have been assigned to a putative new species, Aphytis sp. lingnanensis group. Nevertheless, adult morphology corresponds to A. melinus, which pupa and exuviae are only black in the thorax. For this reason, the current taxonomic status of these individuals (black pupae) remains unclear, allowing us to suggest that it could be a cryptic species of A. melinus. Nowadays, cytochrome c oxidase I (COI) sequence is being considered as a taxonomic character (by Barcoding of Life initiative) and it is used for the description/unveiling of cryptic species. This work presents the use of the DNA barcodes (COI sequence) for the phylogenetic study of the black pupa individuals in the Aphytis group

    Petroleum spray oils and releases of 'aphytis melinus' to control 'aonidiella aurantii' (Maskell) in Spain

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    California red scale, Aonidiella aurantii (Maskell) (Hemiptera: Diaspididae) is one of the main pests of citrus in Spain. Until now the control of this pest has been conducted exclusively with the use of insecticides. The release of parasitoids of the genus Aphytis (Hymenoptera: Aphelinidae) in combination with petroleum spray oils is a biorational control strategy that is successfully applied in other parts of the world. However, this strategy has not been tested in Spain. In this work, we investigated, first the Aphytis species to be released under the Spanish citrus conditions, and second, we conducted preliminary assays to test the efficacy of both, releases of Aphytis melinus DeBach and petroleum spray oils

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