Comparación de la cinética de la infección de ovas de trucha arcoíris (Oncorhynchus mykiss) con dos cepas de Piscirickettsia salmonis detectada mediante dot-blot#

En un estudio previo, se comprobó mediante microscopía de barrido, que la cepa LF-89 se adhiere a la pared de la ovamediante prolongaciones de su membrana externa, estructuras que han sido denominadas "Complejo de AdhesiónPiscirickettsial" o CAP, lo que facilitaría la posterior penetración de la bacteria al interior de la ova. Sin embargo,existen otras cepas aisladas, como la SLGO-95, que es más virulenta y resistente a antibióticos que la LF-89 y que no seha estudiado la posibilidad de unión a ovas. Por lo anterior, en el presente trabajo se comparó la cinética de infección deovas de trucha arcoíris, entre ambas cepas, utilizando como metodología la técnica de dot-blot. El método de "dot-blot"se realizó mediante la retención de proteínas en una membrana de polivinildifluoruro (PVDF), incubación conanticuerpos oligoclonales anti-P. salmonis y posteriormente con un segundo anticuerpo anti inmunoglobulina G deratón conjugado con peroxidasa. La reacción antígeno-anticuerpo se evidenció mediante un sustratoquimioluminiscente, utilizando una película autorradiográfica. La evaluación de la reacción se realizó mediantedensitometría utilizando un "software" computacional. Para estandarizar el método se realizaron diluciones seriadas dela suspensión de P. salmonis, desde 0,02 a 56 μg de proteína total. Para aumentar la sensibilidad se centrifugaron lasmuestras a 11.000 x g por 60 min. Además, se probaron diferentes concentraciones de anticuerpo primario, 1:1000,1:5000 y 1:10.000 y muestras de ovas y bacteria fueron sometidas a desnaturalización por ebullición. Se utilizaron ovasde reproductores libres de infecciones virales, Renibacterium salmoninarum y P. salmonis. Las ovas fueron incubadas(en duplicado) con 500 μL de una suspensión de P. salmonis, ya sea con la cepa LF-89 o SLGO-95 por 1,3, 10 y 60min. Luego, cada ova fue congelada a -70°C hasta ser procesada para la técnica de "dot blot". La muestra nocentrifugada de P. salmonis fue detectada como positiva sólo hasta 0,4 μg de proteína total. Cuando fue centrifugada a11.000 g por 1 h, la bacteria fue detectada hasta la última dilución estudiada (0,02 μg). Por otra parte, no hubo grandesdiferencias en la positividad obtenida mediante las tres diluciones de anticuerpos primarios. De acuerdo, al"background" obtenido y nitidez, se consideró como una mejor dilución de trabajo 1:5000. Cuando se realizó la cinéticade infección con la cepa LF-89, los resultados indicaron la aparición de señal positiva desde 1 min de exposición de lasovas a la bacteria. La reacción positiva se mantuvo hasta los 60 min. Con respecto a la cepa SLGO-95, los resultadosfueron similares, sin embargo, la cepa SLGO-95 demostró un mayor número de píxeles, lo que indica que esta cepa seune a la ova en mayor cantidad. Los resultados apoyan los estudios anteriores que indican que P. salmonis es capaz deinfectar verticalmente las ovas y que esta podría ser una de las formas de transmisión del agente en condiciones decultivo. Además, se comprueba que el grado de infección depende del tipo de cepa actuante.  

Learning Behavioural Context

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Forest hydrology in Chile: Past, present, and future

This paper reviews the current knowledge of hydrological processes in Chilean temperate forests which extend along western South America from latitude 29° S to 56 ° S. This geographic region includes a diverse range of natural and planted forests and a broad sweep of vegetation, edaphic, topographic, geologic, and climatic settings which create a unique natural laboratory. Many local communities, endangered freshwater ecosystems, and downstream economic activities in Chile rely on water flows from forested catchments. This review aims to (i) provide a comprehensive overview of Chilean forest hydrology, to (ii) review prior research in forest hydrology in Chile, and to (iii) identify knowledge gaps and provide a vision for future research on forest hydrology in Chile. We reviewed the relation between native forests, commercial plantations, and other land uses on water yield and water quality from the plot to the catchment scale. Much of the global understanding of forests and their relationship with the water cycle is in line with the findings of the studies reviewed here. Streamflow from forested catchments increases after timber harvesting, native forests appear to use less water than plantations, and streams draining native forest yield less sediment than streams draining plantations or grassland/shrublands. We identified 20 key knowledge gaps such as forest groundwater systems, soil–plant-atmosphere interactions, native forest hydrology, and the effect of forest management and restoration on hydrology. Also, we found a paucity of research in the northern geographic areas and forest types (35-36 ° S); most forest hydrology studies in Chile (56 %) have been conducted in the southern area (Los Rios Region around 39-40 ° S). There is limited knowledge of the geology and soils in many forested areas and how surface and groundwater are affected by changes in land cover. There is an opportunity to advance our understanding using process-based investigations linking field studies and modeling. Through the establishment of a forest hydrology science “society” to coordinate efforts, regional and national-scale land use planning might be supported. Our review ends with a vision to advance a cross-scale collaborative effort to use new nation-wide catchment-scale networks Long-term Ecosystem Research (LTER) sites, to promote common and complementary techniques in these studies, and to conduct transdisciplinary research to advance sound and integrated planning of forest lands in Chile

Proactive control of cresting in homogeneous oil reservoirs : an experimental study

Cresting in horizontal wells is a well-known reservoir problem usually described as the insurgence of effluent(s) (unwanted water and or gas) through the perforation of the well, which is produced together with oil. Cresting is majorly affected by pressure drop, resulting in uneconomic oil production rates and large volumes of oil could be left behind due to premature shut-in of the well. This study experimentally investigates the use of electromagnetic valve in proactively controlling production of water during cresting from homogeneous thick- and thin-oil rim reservoirs, based on the principle of capillarity (reservoir wettability) and effluents (water and gas) breakthrough time. A time, half the approximated initial effluents breakthrough time, was pre-set for the electromagnetic valve to close. The valve closed almost immediately at the set-time thereby shutting oil production temporarily, causing the water and gas height levels to recede by gravity and capillarity. The efficiency of this technique was compared with an uncontrolled simulation case, in terms of cumulative oil produced and water produced at the same overall production time. Using the cresting control procedure, higher percentages in oil produced and water reduction were observed in the cases controlled proactively. An increment of 3.56% in oil produced and decrement in cumulative water produced of 9.96% were observed for the thick-oil rim reservoir while little increment in oil produced of 0.7% and lower water reduction of 1.03% were observed in the thin-oil rim reservoir. Hence, the effectiveness of the cresting control procedure depends on the oil-column height in the reservoir

Patch-level spatial layout for classification and weakly supervised localization

International audienceWe propose a discriminative patch-level spatial layout model suitable for training with weak supervision. We start from a block-sparse model of patch appearance based on the normalized Fisher vector representation. The appearance model is responsible for i) selecting a discriminative subset of visual words, and ii) identifying distinctive patches assigned to the selected subset. These patches are further filtered by a sparse spatial model operating on a novel representation of pairwise patch layout. We have evaluated the proposed pipeline in image classification and weakly supervised localization experiments on a public traffic sign dataset. The results show significant advantage of the proposed spatial model over state of the art appearance models