Los autores desean agradecer a todos los pescadores de langostas comerciales de Nueva Inglaterra cuyos barcos se nos permitió muestra de - su cooperación es muy apreciada. Además, agradecemos a Little Bay Lobster Company (Newington, Nueva Hampshire) y de Champlin Mariscos (Point Judith, Rhode Island), que nos permitió realizar el muestreo en sus instalaciones. Los siguientes estudiantes UNH proporcionaron asistencia invaluable a lo largo de este estudio: Haley Blanco, Françoise Morrison, Sarah Havener, Audra Chaput, y mayo Grose. Este proyecto fue apoyado por becas de NH SeaGrant a WHW (proyecto # R/CFR-11) y una subvención del programa UNH Marino que JSG.

Parte A: El campo de muestreo Técnica 1. Medidas Femenino <ol><li> Para cada langosta hembra, medir la longitud del caparazón (CL) y la anchura del segundo segmento abdominal para los 1,0 mm más próximo utilizando un par de pinzas. NOTA: El segundo segmento abdominal porque se mide la anchura del abdomen (donde las mujeres llevan sus huevos) es un indicador de la madurez sexual. Además, podría ser ventajoso para determinar la etapa de muda para que coincida más adelante estos datos con la etapa reproductiva (véase el paso 1.2). </li><li> Clip una pequeña porción distal de uno de los pleópodos abdominales (un protocolo metodológico estándar de langostas) y almacenar el pleópodo en agua de mar limpia para su posterior visualización con un microscopio de disección 9. NOTA: Este paso también puede revelar la presencia de glándulas del cemento en los pleópodos, que son una indicación de que las mujeres se están preparando para la extrusión de los huevos 10. </li></ol> 2. Aguja InseRTion <ol><li> Inserte la aguja de la jeringa (18 G, véase el cuadro 1) ~ 1 cm en el receptáculo seminal, en ángulo hacia la cola en el ~ 45 ° (Figura 1). NOTA: En langostas con tapones de espermatozoides intactos, la aguja puede satisfacer cierta resistencia durante la inserción, ya que se empuja a través del tapón de esperma antes de alcanzar la más alta concentración de espermatozoides en la parte inferior del receptáculo seminal (Figura 1). NOTA: La presencia de la clavija, en sí mismo, es un indicador fiable de que las langostas se han apareado. En algunos animales puede haber poca resistencia, posiblemente porque no espermatóforo está presente, el espermatóforo es pequeño, no ha cristalizado en un enchufe todavía, o es posible que ya se han utilizado para fertilizar una o varias puestas de huevos. </li></ol><img alt="Figura 1" fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig1highres.jpg" src="/files/ftp_upload/50498/50498fig1.jpg" width = &quot;600px&quot; /&gt; Figura 1. La inserción de la aguja y la vista de la langosta receptáculo seminal. Inserción de la aguja en el receptáculo seminal de la langosta de punta roma para extraer una muestra de esperma. El ángulo y la profundidad de la aguja son tanto crítico para la obtención de muestras consistentes. Derecha: Vista de un receptáculo seminal que ha sido diseccionada y dividida verticalmente. El tapón de esperma ocupa la parte superior o exterior del receptáculo, mientras que los espermatozoides se encuentra en la parte inferior (el punto más profundo) del receptáculo. Es importante tener en cuenta que el muestreo por debajo de esta profundidad puede perforar el receptáculo y el resultado en una muestra contaminada con sangre. <a href="https://www.jove.com/files/ftp_upload/50498/50498fig1highres.jpg" target="_blank">Haga clic aquí para ver la imagen más grande</a> . 3. Penetrar el esperma Plug <ol><li> Después de la inserción inicial, más pronunciada el ángulo de la aguja a casi vertical (perpendicular al eje del cuerpo), y muy lentamente comenzar a trabajar sehacia abajo en el receptáculo. </li><li> Mantenga la aguja inclinada en un ángulo tal que la punta está apuntando ligeramente hacia la cola. En este caso, la punta de la aguja está comprimiendo la parte superior del tapón, hasta que eventualmente penetra en la matriz. </li><li> Continuar para empujar la aguja lentamente hacia abajo hasta sentir una resistencia, una vez más - esta es la parte inferior del recipiente y donde se toma la muestra. </li></ol> 4. Extracción de esperma <ol><li> Extraer una muestra (a menudo material del tapón sólido, líquido y espermatozoides otro fluido) desde la parte inferior del receptáculo y depositar la muestra en un tubo de plástico de 2,0 ml etiquetados. </li><li> Enjuague el recipiente con ~ 0,1 a 0,5 ml de agua de mar fría. </li><li> Retire aproximadamente 0,3 ml de líquido y coloque en el mismo tubo. </li><li> Muestras de esperma tienda en hielo hasta que puedan ser examinados en el laboratorio. Parte B: El examen de las muestras de semen en el laboratorio NOTA: Las muestras extraídas de los receptáculos seminales de l individuo<html> obsters deben ser examinados para determinar si los espermatozoides estaban presentes o ausentes. </li><li> Retire ~ 50 l de fluido de cada tubo de plástico, lugar en un portaobjetos de vidrio con una hoja de cubierta, y vista a 100X con un microscopio compuesto. </li><li> Puntuación muestras como: esperma escaso, numerosas, o ausentes (este estudio no cuantificó el número de espermatozoides reales, Figura 2). </li></ol><img alt="Figura 2" fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig2highres.jpg" src="/files/ftp_upload/50498/50498fig2.jpg" width="600px" /> Figura 2. Imágenes de muestra de esperma de langosta. Imágenes de las muestras de semen de langosta tomadas de receptáculo seminal de una mujer, tal como se observa con un microscopio compuesto a 40X de aumento. La imagen en la izquierda muestra una muestra que contenía numerosos espermatozoides mientras que el otro (derecha) ilustra una muestra escasa (barra de escala = 10 micras).. &quot;Target =&quot; _blank jpg &quot;&gt; Haga clic aquí para ver la imagen más grande.

<ol>
	<li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> FAO and Agriculture Organization of the United Nations). Fisheries and Aquaculture Department. , (2012).</li><li>MacDiarmid, A., Butler, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sperm+economy+and+limitation+in+spiny+lobsters.">Sperm economy and limitation in spiny lobsters.</a> Behav. Ecol. Sociobiol. 46, 14-24 (1999).</li><li>Kendall, M. S., Wolcott, D. L., Wolcott, T. G., Hines, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+male+size+and+mating+history+on+sperm+content+of+ejaculates+of+the+blue+crab+Callinectes+sapidus.">Influence of male size and mating history on sperm content of ejaculates of the blue crab Callinectes sapidus.</a> 230, 235-240 (2002).</li><li>Sato, T., Ashidate, M., Goshima, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+male+mating+frequency+and+male+size+on+ejaculate+size+and+reproductive+success+of+female+spiny+king+crab,+Parolithodes+brevipes.">Effects of male mating frequency and male size on ejaculate size and reproductive success of female spiny king crab, Parolithodes brevipes.</a> Mar. Ecol. Prog. Ser. 296, 251-262 (2005).</li><li>Sato, T., Goshima, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impacts+of+male-only+fishing+and+sperm+limitation+in+manipulated+populations+of+an+unfished+crab,+Hapalogaster.">Impacts of male-only fishing and sperm limitation in manipulated populations of an unfished crab, Hapalogaster.</a> 313, 193-204 (2006).</li><li>Rondeau, A., Sainte-Marie, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variable+mate-guarding+time+and+sperm+allocation+by+male+snow+crabs+(Chionoecetes+opilio)+in+response+to+sexual+competition,+and+their+impact+on+the+mating+success+of+females.">Variable mate-guarding time and sperm allocation by male snow crabs (Chionoecetes opilio) in response to sexual competition, and their impact on the mating success of females.</a> Biol. Bull. 201, 204-217 (2001).</li><li>Waddy, S. L., Aiken, D. E., Bauer, R. T., Martin, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Mating and insemination in the Ameri can lobster,Homarus americanus. Crustacean sexual behavior. , 126-144 (1990).</li><li>Johnson, K. J., Goldstein, J. S., Watson, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+methods+for+determining+the+fertility+status+in+early-stage+American+lobster,+Homarus+americanus,+eggs.">Two methods for determining the fertility status in early-stage American lobster, Homarus americanus, eggs.</a> J. Crust. Biol. 31, 693-700 (2011).</li><li>Aiken, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proecdysis,+setal+development,+and+molt+prediction+in+the+American+lobster+(Homarus).">Proecdysis, setal development, and molt prediction in the American lobster (Homarus).</a> 30, 1337-1344 (1973).</li><li>Aiken, D. E., Waddy, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cement+gland+development,+ovary+maturation,+and+reproductive+cycles+in+the+American+lobster+Homarus+americanus.">Cement gland development, ovary maturation, and reproductive cycles in the American lobster Homarus americanus.</a> J. Crust. Biol. 2, 315-327 (1982).</li><li>Pugh, T. L., Goldstein, J. S., Lavalli, K. L., Clancy, M., Watson, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=At-sea+determination+of+female+American+lobsters+(Homarus+americanus)+mating+activity:+Patterns+vs.+expectations.">At-sea determination of female American lobsters (Homarus americanus) mating activity: Patterns vs. expectations.</a> Fish. Res. 147, 327-337 (2013).</li><li>Bauer, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phylogenetic+trends+in+sperm+transfer+and+storage+complexity+in+decapod+crustaceans.">Phylogenetic trends in sperm transfer and storage complexity in decapod crustaceans.</a> J. Crust. Biol. 6, 313-325 (1986).</li><li>Talbot, P., Helluy, S., Factor, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproduction+and+embryonic+development.">Reproduction and embryonic development.</a> Biology of the Lobster: Homarus americanus. , 177-216 (1995).</li><li>Cooper, R. A., Uzmann, J. R., Cobb, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ecology of juvenile and adult Homarus. The biology and management of lobsters. 2, 97-142 (1980).</li><li>Estrella, B. T., McKiernan, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catch-per-unit-effort+and+biological+parameters+from+the+Massachusetts+coastal+lobster+(Homarus+americanus)+resource:+description+and+trends.">Catch-per-unit-effort and biological parameters from the Massachusetts coastal lobster (Homarus americanus) resource: description and trends.</a> NOAA Tech. Rep. NMFS. 81, (1989).</li><li>Waddy, S. L., Aiken, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+fertilization+and+consecutive+spawning+in+large+American+lobster,+Homarus+americanus.">Multiple fertilization and consecutive spawning in large American lobster, Homarus americanus.</a> Can. J. Fish. Aquat. Sci. 43, 2291-2294 (1986).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atlantic+States+Marine+Fisheries+Commission+(ASMFC).+American+lobster+stock+assessment+report+for+peer+review.">Atlantic States Marine Fisheries Commission (ASMFC). American lobster stock assessment report for peer review.</a> ASMFC Stock Assessment. , (2009).</li></ol>

Los autores declaran no tener ningún conflicto de intereses.

Este esquema de vídeo y papel y demostrar un método para determinar si langostas hembra se han apareado con éxito. Este enfoque es bastante simple que se puede utilizar para la toma de muestras a gran escala de langostas en el mar por una variedad de usuarios. El método se basa en la detección de la presencia de un espermatóforo en el receptáculo seminal que se transmite de hombre a langostas hembra durante el apareamiento. Estos espermatóforos endurecen parcialmente en un tapón de esperma poco después de que ...

<html> Langosta americana (Homarus americanus) es una de las pesquerías más productivas del Atlántico Norte (~ 56 000 toneladas en 2011, valorado en más de $ 390 millones) 1. Sin embargo, hay una falta general de comprensión de la dinámica de reproducción de esta especie en las poblaciones silvestres. La generación de estimaciones más precisas de producción reproductiva, incluyendo el número y tamaño de los individuos que participan activamente en la reproducción, se puede mejorar el proceso de evaluación de la población. Por ejemplo, las mujeres que se les impide alcanzar su potencial reproductivo completo debido a la limitación de esperma se han identificado como una preocupación para varios crustáceos marinos explotados comercialmente, incluyendo: langostas, cangrejos azules 2 3, 4 cangrejos rey, cangrejos de piedra 5, y los cangrejos de nieve 6. El objetivo general es determinar si la limitación de esperma podría ser un factor en algunas regiones de la pesquería de la langosta americana. nt &quot;&gt; Se observaron los primeros signos de un problema potencial limitación de esperma en langostas, mientras que la realización de un estudio de marcado no relacionada con las langostas hembras ovígeras (con huevos visiblemente intactos, recientemente extruidos). recaptura informes de los pescadores indicó que ~ 15% de estos animales había disminuido sus puestas de huevos después de sólo 1-2 meses. La hipótesis de trabajo fue que algunas langostas estaban llevando los huevos que no habían sido fertilizados y, como resultado, &quot;se cayó&quot; 7. Los datos preliminares de un estudio posterior ha confirmado el hecho de que las langostas se extrusión de huevos, incluso si no se han apareado con éxito y estos huevos no fertilizados sólo se llevan durante ~ 1 meses 8. Por lo tanto, teniendo en cuenta la observación de las hembras llevan los huevos no fertilizados en las poblaciones naturales, hemos tratado de determinar el grado en el cual la prescripción de esperma podría estar contribuyendo a un nivel submáximo de reproducción por las langostas sexualmente maduros. Para lograr este objetivo, se desarrolló una técnica para detectar la espermatogénesisphore depositada por los machos durante el apareamiento. Este documento y el vídeo se describen un método no invasivo, simple desarrollada para determinar el éxito de apareamiento de las langostas hembras. La técnica se puede utilizar de forma rápida y fiable en el mar, ya sea a bordo de la investigación o de buques pesqueros comerciales. Los detalles de este método de muestreo, así como algunos resultados representativos se presentan para ilustrar la aplicación de la técnica.

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a noninvasive method for in situ determination of mating success in female american lobsters (homarus americanus)

A pesar de ser una de las pesquerías más productivas del Atlántico Noroeste, aún queda mucho por conocer acerca de la dinámica reproductiva naturales de langostas americanas. Trabajos recientes en las poblaciones de crustáceos explotados (cangrejos y langostas) sugiere que hay circunstancias en las hembras maduras son incapaces de alcanzar su potencial reproductivo completo debido a la limitación de los espermatozoides. Para examinar esta posibilidad en diferentes regiones de la pesquería de langosta de América, un método confiable y no invasivo fue desarrollado para el muestreo de un gran número de langostas hembras en el mar. Este método consiste en insertar una aguja en el receptáculo seminal de la hembra de punta roma para determinar la presencia o ausencia de un tapón de esperma y para extraer una muestra que puede ser examinada para detectar la presencia de espermatozoides. Una serie de estudios de control se llevaron a cabo en el muelle y en el laboratorio para poner a prueba la fiabilidad de esta técnica. Estos esfuerzos implicaron muestreo 294 langostas hembra para confirmar que la presencia oenchufe de esperma FA era un indicador fiable de los espermatozoides dentro del receptáculo y, por tanto, el apareamiento. Este documento detalla la metodología y los resultados obtenidos a partir de un subconjunto de las hembras total de la muestra. De los 230 langostas hembra muestreados de Banco de George y Cape Ann, MA (rango 71-145 size = mm de longitud del caparazón), el 90,3% fueron positivas para el esperma. Las posibles explicaciones para la ausencia de espermatozoides en algunas mujeres incluyen: falta de madurez (falta de madurez fisiológica), la ruptura del tapón de esperma después de ser utilizado para fertilizar una nidada de huevos, y la falta de actividad de apareamiento. Las encuestas indican que esta técnica para examinar el éxito de apareamiento de langostas hembra es un proxy fiable que se puede utilizar en el campo para documentar la actividad reproductiva en las poblaciones naturales.

<html> Los datos aquí presentados son para un subconjunto de las langostas hembras que se originaron en tanto cerca de la costa y de los centros extraterritoriales. El grupo constaba de 44 nearshore langostas que fueron capturadas en las aguas costeras cerca de Cape Ann, Massachusetts y transportan en neveras al laboratorio para su análisis. Se capturaron un total de 186 langostas en alta mar en el Banco de George y se tomaron muestras a un centro de detención comercial en Nueva Hampshire. ...

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A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

Los cambios inducidos por la pesca-para la pesca de crustáceos explotados, como la pesca de la langosta americana, potencialmente podrían influir en su dinámica reproductiva, lo que lleva a una reducción en el éxito de apareamiento. El objetivo de este estudio fue desarrollar un método no invasivo para determinar el éxito de apareamiento de las langostas hembras que pueden ser fisiológicamente o funcionales madura.

Un método no invasivo para<em&gt; In situ</em&gt; Determinación de Éxito de apareamiento en americano femenino Langostas (<em&gt; Homarus americanus</em&gt;)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American ...

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A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

10.9K vistas.  University of New Hampshire.  Los cambios inducidos por la pesca-para la pesca de crustáceos explotados, como la pesca de la langosta americana, potencialmente podrían influir en su dinámica reproductiva, lo que lleva a una reducción en el éxito de apareamiento. El objetivo de este estudio fue desarrollar un método no invasivo para determinar el éxito de apareamiento de las langostas hembras que pueden ser fisiológicamente o funcionales madura. 

Video: A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

A Technique to Screen American Beech for Resistance to the Beech Scale Insect (Cryptococcus fagisuga Lind.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The disease is initiated by feeding activities of the invasive beech scale insect, Cryptococcus fagisuga, which creates entry points for infection by one of the Neonectria species of fungus.&#160;Without scale infestation, there is little opportunity for fungal infection. Using scale eggs to artificially infest healthy trees in heavily BBD impacted stands demonstrated that these trees were resistant to the scale insect portion of the disease complex1.&#160;Here we present a protocol that we have developed, based on the artificial infestation technique by Houston2, which can be used to screen for scale-resistant trees in the field and in smaller potted seedlings and grafts.&#160;The identification of scale-resistant trees is an important component of management of BBD through tree improvement programs and silvicultural manipulation.

A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind.

Beech bark disease is initiated by feeding activities of the beech scale insect that create fungal entry points in the bark.&#160;Trees that are resistant to the scale insect are also disease resistant.&#160;Here we present the protocol we have developed to screen individual beech trees for beech scale resistance.

Una técnica a la pantalla haya americana para la resistencia al insecto Haya Scale (<em&gt; Fagisuga Cryptococcus</em&gt; Lind.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The ...

Investigación

Medio ambiente

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

VacuSIP, un método mejorado para InEx<em&gt; In Situ</em&gt; Medición de las partículas y compuestos disueltos procesada por Alimentadores Active Suspension

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

Optimized Procedure for Determining the Adsorption of Phosphonates onto Granular Ferric Hydroxide using a Miniaturized Phosphorus Determination Method

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide (GFH), with little effort and high reliability. The phosphonate, e.g., nitrilotrimethylphosphonic acid (NTMP), is brought into contact with the GFH in a rotator in a solution buffered by an organic acid (e.g., acetic acid) or Good buffer (e.g., 2-(N-morpholino)ethanesulfonic acid) [MES] and N-cyclohexyl-2-hydroxyl-3-aminopropanesulfonic acid [CAPSO]) in a concentration of 10 mM for a specific time in 50 mL centrifuge tubes. Subsequently, after membrane filtration (0.45 &#181;m pore size), the total phosphorus (total P) concentration is measured using a specifically developed determination method (ISOmini). This method is a modification and simplification of the ISO 6878 method: a 4 mL sample is mixed with H2SO4 and K2S2O8 in a screw cap vial, heated to 148-150 &#176;C for 1 h and then mixed with NaOH, ascorbic acid and acidified molybdate with antimony(III) (final volume of 10 mL) to produce a blue complex. The color intensity, which is linearly proportional to the phosphorus concentration, is measured spectrophotometrically (880 nm). It is demonstrated that the buffer concentration used has no significant effect on the adsorption of phosphonate between pH 4 and 12. The buffers, therefore, do not compete with the phosphonate for adsorption sites. Furthermore, the relatively high concentration of the buffer requires a higher dosage concentration of oxidizing agent (K2S2O8) for digestion than that specified in ISO 6878, which, together with the NaOH dosage, is matched to each buffer. Despite the simplification, the ISOmini method does not lose any of its accuracy compared to the standardized method.

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide, with little effort and high reliability. In a buffered solution, the phosphonate is brought into contact with the adsorbent using a rotator and then analyzed via a miniaturized phosphorus determination method.

Procedimiento optimizado para la determinación de la adsorción de los fosfonatos en hidróxido férrico Granular utilizando un método de determinación de fósforo miniaturizados

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric ...

In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples

Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ocean temperatures and ocean acidification, corals are increasingly under threat. To tackle these challenges, advances in cell and molecular biology have proven to be crucial for diagnosing the health of corals. Modifying some of the techniques commonly used in human medicine could greatly improve researchers&#39; ability to treat and save corals. To address this, a protocol for in situ hybridization used primarily in human medicine and evolutionary developmental biology has been adapted for use in adult corals under stress.
The purpose of this method is to visualize the spatial expression of an RNA probe in adult coral tissue that has been embedded in paraffin and sectioned onto glass slides. This method focuses on removal of the paraffin and rehydration of the sample, pretreatment of the sample to ensure permeability of the sample, pre-hybridization incubation, hybridization of the RNA probe, and visualization of the RNA probe. This is a powerful method when using non-model organisms to discover where specific genes are expressed, and the protocol can be easily adapted for other non-model organisms. However, the method is limited in that it is primarily qualitative, because expression intensity can vary depending on the amount of time used during the visualization step and the concentration of the probe. Furthermore, patience is required, as this protocol can take up to 5 days (and in many cases, longer) depending on the probe being used. Finally, non-specific background staining is common, but this limitation can be overcome.

In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples

The goal of this protocol is to perform in situ hybridization on adult coral samples that have been embedded in paraffin and sectioned onto glass slides. This is a qualitative method used to visualize the spatial expression of an RNA anti-sense probe in paraffin-embedded tissues.

In Situ Técnicas de hibridación para muestras de corales adultos parafinadas

Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ...

A Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Identification of Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many countries around the world. Severe yield losses are caused by direct feeding, and even more importantly, also by the transmission of more than 100 harmful plant pathogenic viruses. As for other invasive pests, increased international trade facilitates the dispersal of B. tabaci to areas beyond its native range. Inspections of plant import products at points of entry such as seaports and airports are, therefore, seen as an important prevention measure. However, this last line of defense against pest invasions is only effective if rapid identification methods for suspicious insect specimens are readily available. Because the morphological differentiation between the regulated B. tabaci and close relatives without quarantine status is difficult for non-taxonomists, a rapid molecular identification assay based on the loop-mediated isothermal amplification (LAMP) technology has been developed. This publication reports the detailed protocol of the novel assay describing rapid DNA extraction, set-up of the LAMP reaction, as well as interpretation of its read-out, which allows identifying B. tabaci specimens within one hour. Compared to existing protocols for the detection of specific B. tabaci biotypes, the developed method targets the whole B. tabaci species complex in one assay. Moreover the assay is designed to be applied on-site by plant health inspectors with minimal laboratory training directly at points of entry. Thorough validation performed under laboratory and on-site conditions demonstrates that the reported LAMP assay is a rapid and reliable identification tool, improving the management of B. tabaci.

A Loop-mediated Isothermal Amplification LAMP Assay for Rapid Identification of Bemisia tabaci

This paper reports the protocol for a rapid identification assay for Bemisia tabaci based on loop-mediated isothermal amplification (LAMP) technology. The protocol requires minimal laboratory training and can, therefore, be implemented on-site at points of entry for plant imports such as seaports and airports.

Un ensayo de amplificación isotérmica (lámpara) mediada por el lazo para identificación rápida de Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many ...

Implantation of a New Micro Acoustic Tag in Juvenile Pacific Lamprey and American Eel

Juvenile Pacific Lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology has been identified as a way to obtain more detailed information on movement and behavior across a broader spatial scale than is possible with other known technology. The purpose of this method is to provide a detailed step by step instruction on tag implantation for both lampreys and eel. For laboratory studies using actively migrating juvenile Pacific Lamprey (120-160 mm), we determined that the presence of the tag did not alter the swimming ability between tagged and untagged Individuals or have any significant tag loss (&lt;3%). Similar results were determined during laboratory testing of Yellow phase American Eels (113-175 mm). No mortality occurred during a 38-day holding period and there was minimal tag loss (3.8%). The presence of the tag did not have any significant effect on the swimming ability or survival of tagged eels compared to untagged controls and there was minimal tag loss.

This article illustrates the procedure for implanting a micro acoustic tag in juvenile Pacific Lamprey and American Eel. Our application is based on laboratory trials, which indicate that acoustic tags can be successfully implanted with no apparent effects on swimming ability or survival and minimal tag loss.

Implantación de una nueva etiqueta acústica Micro en juvenil Pacífico lamprea y anguila americana

Juvenile Pacific Lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology ...

A Telemetric, Gravimetric Platform for Real-Time Physiological Phenotyping of Plant&ndash;Environment Interactions

Food security for the growing global population is a major concern. The data provided by genomic tools far exceeds the supply of phenotypic data, creating a knowledge gap. To meet the challenge of improving crops to feed the growing global population, this gap must be bridged.
Physiological traits are considered key functional traits in the context of responsiveness or sensitivity to environmental conditions. Many recently introduced high-throughput (HTP) phenotyping techniques are based on remote sensing or imaging and are capable of directly measuring morphological traits, but measure physiological parameters mainly indirectly.
This paper describes a method for direct physiological phenotyping that has several advantages for the functional phenotyping of plant&#8211;environment interactions. It helps users overcome the many challenges encountered in the use of load-cell gravimetric systems and pot experiments. The suggested techniques will enable users to distinguish between soil weight, plant weight and soil water content, providing a method for the continuous and simultaneous measurement of dynamic soil, plant and atmosphere conditions, alongside the measurement of key physiological traits. This method allows researchers to closely mimic field stress scenarios while taking into consideration the environment&#8217;s effects on the plants&#8217; physiology. This method also minimizes pot effects, which are one of the major problems in pre-field phenotyping. It includes a feed-back fertigation system that enables a truly randomized experimental design at a field-like plant density. This system detects the soil-water-content limiting threshold (&#952;) and allows for the translation of data into knowledge through the use of a real-time analytic tool and an online statistical resource. This method for the rapid and direct measurement of the physiological responses of multiple plants to a dynamic environment has great potential for use in screening for beneficial traits associated with responses to abiotic stress, in the context of pre-field breeding and crop improvement.

A Telemetric, Gravimetric Platform for Real-Time Physiological Phenotyping of Plant&#8211;Environment Interactions

This high-throughput, telemetric, whole-plant water relations gravimetric phenotyping method enables direct and simultaneous real-time measurements, as well as the analysis of multiple yield-related physiological traits involved in dynamic plant&#8211;environment interactions.

Una plataforma telemétrica y gravimétrica para el fenotipado fisiológico en tiempo real de las interacciones entre plantas y entornos

Food security for the growing global population is a major concern. The data provided by genomic tools far exceeds the supply of phenotypic data, creating ...

Mistletoe Eradicator - A Novel Tool for Simultaneous Mechanical and Chemical Control of Mistletoe

Some species of mistletoes, an aerial plant hemiparasites, play a Janus-like role in ecosystems by being rich biodiversity hotspots due to their trophic and topic associations with various organisms and at the same time acting as biological pirates, exhausting the host trees by stealing water and nutrients via the haustorium. Many crop and timber trees are attacked by mistletoes, for example, dwarf mistletoes parasitize conifers, representatives of genus Tapinanthus target guava, and Phoradendron mistletoe target pecans. Recently, an increasing infestation of the European mistletoe (Viscum album subsp. album L.) in monoculture stands of common walnuts (Juglans regia L.)&#160;was recorded for the first time during 2018 in different regions of Kashmir Valley, India. The common chemical approach based on the spraying of mistletoe exophytes with the growth regulators, ethephon and glyphosate, was not efficient since V. album resprouted in the next vegetation season. Hand pruning of mistletoe was time-consuming, and a polyethylene covering resulted in the localized rotting of host tissues. The most efficient approach to eradicate mistletoe was the use of a novel tool called Mistletoe Eradicator, which allows to use mechanical and chemical means of mistletoe control simultaneously. It consists of a manipulating pole with the terminal pruning saw having a container filled with the growth regulator, from which the liquid slowly drops onto the saw blade through the nozzle. It allows working both from the ground and in the tree canopy. After the targeted application of 5%-10% ethephon or glyphosate to the base of the just pruned V. album stems (stubs), exophyte resprouting was significantly reduced 9 months after the pruning as compared to the only pruned or sprayed exophytes. An alternative variant of Mistletoe Eradicator was constructed in Ukraine filled just with a food color additive to track the liquid dropping and was successfully tested.

The protocol describes an easy-to-handle tool called Mistletoe Eradicator which combines mechanical and chemical approaches and is efficient in the control of mistletoe infestation on the host trees. The manuscript uses the European mistletoe (Viscum album subsp. album L.) as an example.

Erradicador de muérdago - Una herramienta novedosa para el control mecánico y químico simultáneo del muérdago

Some species of mistletoes, an aerial plant hemiparasites, play a Janus-like role in ecosystems by being rich biodiversity hotspots due to their trophic ...

In Situ Soil Moisture Sensors in Undisturbed Soils

Soil moisture directly affects operational hydrology, food security, ecosystem services, and the climate system. However, the adoption of soil moisture data has been slow due to inconsistent data collection, poor standardization, and typically short record duration. Soil moisture, or quantitatively volumetric soil water content (SWC), is measured using buried, in situ sensors that infer SWC from an electromagnetic response. This signal can vary considerably with local site conditions such as clay content and mineralogy, soil salinity or bulk electrical conductivity, and soil temperature; each of these can have varying impacts depending on the sensor technology.
Furthermore, poor soil contact and sensor degradation can affect the quality of these readings over time. Unlike more traditional environmental sensors, there are no accepted standards, maintenance practices, or quality controls for SWC data. As such, SWC is a challenging measurement for many environmental monitoring networks to implement. Here, we attempt to establish a community-based standard of practice for in situ SWC sensors so that future research and applications have consistent guidance on site selection, sensor installation, data interpretation, and long-term maintenance of monitoring stations.
The videography focuses on a multi-agency consensus of best-practices and recommendations for the installation of in situ SWC sensors. This paper presents an overview of this protocol along with the various steps essential for high-quality and long-term SWC data collection. This protocol will be of use to scientists and engineers hoping to deploy a single station or an entire network.

In Situ Soil Moisture Sensors in Undisturbed Soils

The determination of soil water content is a critical mission requirement for many state and federal agencies. This protocol synthesizes multi-agency efforts to measure soil water content using buried in situ sensors.

In situ Sensores de humedad del suelo en suelos no perturbados

Soil moisture directly affects operational hydrology, food security, ecosystem services, and the climate system. However, the adoption of soil moisture ...

A Low-Cost Method of Measuring the In Situ Primary Productivity of Periphyton Communities of Lentic Waters

Measuring the in situ primary productivity of periphyton during the growing season gradient can elucidate the quantitative effect of environmental drivers (mainly phosphorus concentration and light intensity) and species composition on primary productivity. Primary productivity is mainly driven by light intensity, temperature, availability of nutrients, and distribution of the ionic species of the carbonate system in the respective depths of the euphotic zone. It is a complex system that is very difficult to simulate in the laboratory. This cheap, transportable, and easy-to-build floating barge allows measuring the primary productivity accurately-directly under the actual natural conditions. The methodology is based on measuring the primary productivity in real time using noninvasive oxygen sensors integrated into tightly sealed glass jars, enabling online oxygen flux monitoring and providing new insights into metabolic activities. Detailed seasonal in situ measurements of gross primary productivity of microbial mats (or other benthic organisms) can improve current knowledge of the processes controlling primary productivity dynamics in lentic waters.

A Low-Cost Method of Measuring the In Situ Primary Productivity of Periphyton Communities of Lentic Waters

Presented here is a cost-effective and transportable method/facility for measuring the primary productivity of microbial mats under actual in situ environmental temperature and light conditions. The experimental setup is based on widely available materials and can be used under various conditions while offering the advantages of laboratory-based models.

Un método de bajo costo para medir la productividad primaria in situ de las comunidades de perifitón de las aguas lénticas

Measuring the in situ primary productivity of periphyton during the growing season gradient can elucidate the quantitative effect of environmental drivers ...