Die Autoren möchten all die New England kommerziellen Hummerfischer, deren Boote durften wir aus probieren danken - ihre Zusammenarbeit wird sehr geschätzt. Außerdem bedanken wir uns bei Little Bay Lobster Company (Newington, NH) und Champlin Seafood (Point Judith, RI), die uns die Probenahme an ihren Standorten zu führen erlaubt. Die folgenden UNH Studenten unschätzbare Unterstützung während dieser Studie: Haley Weiß, Françoise Morrison, Sarah Havener, Audra Chaput und Mai Grose. Dieses Projekt wurde durch Zuschüsse aus NH SeaGrant zu WHW (Projekt # R/CFR-11) und eine UNH Marine Programm Grant JSG unterstützt.

Teil A: Feldprobenahmetechnik 1. Weiblich Messungen <ol><li> Für jede weibliche Hummer, messen die Panzerlänge (CL) und der Breite des zweiten Bauchsegment auf die nächsten 1,0 mm mit einem Paar von Bremssätteln. HINWEIS: Der zweite Bauch Segment gemessen, da die Breite der Bauch (wo Weibchen tragen ihre Eier) ist ein Indikator für die Geschlechtsreife. Darüber hinaus könnte es vorteilhaft sein, um die Häutung Stadium festzustellen, um später diese Daten mit der reproduktiven Phase übereinstimmen (siehe Schritt 1.2). </li><li> Clip einen kleinen distalen Teil von einem der Bauch pleopods (ein Standard-Protokoll in methodischen Hummer) und speichern Sie die Pleopoden in sauberem Meerwasser für die spätere Ansicht mit einem Binokular 9. HINWEIS: Dieser Schritt könnte auch zeigen das Vorhandensein von Zement Drüsen an den Pleopoden, die ein Indiz, dass Frauen bereiten, um ihre Eier 10 extrudieren sind. </li></ol> 2. Needle Insertion <ol><li> Legen Sie die Spritzennadel (18 G, siehe Tabelle 1) ~ 1 cm in die Samenbehälter, gewinkelt in Richtung der Schwanz bei ~ 45 ° (Abbildung 1). HINWEIS: In Hummer mit intakten Spermien Stecker, kann die Nadel einen Widerstand beim Einsetzen zu erfüllen, wie sie durch die Spermien Stecker vor dem Erreichen der höchsten Konzentration von Spermien an der Unterseite der Samenbehälter (1) geschoben. HINWEIS: Die Anwesenheit der Stecker, in sich selbst, ist ein zuverlässiger Indikator dafür, dass Hummer haben gepaart. Bei einigen Tieren kann es zu wenig Widerstand, möglicherweise, weil kein Spermatophore vorhanden ist, ist die Spermatophore klein, hat es nicht in einen Stecker noch gehärtet, oder es kann bereits verwendet wurden, um eine oder mehrere Gelege zu befruchten. </li></ol><img alt="Figur 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; Fig. 1 ist. Nadeleinführung und Blick Hummer Samenbehälter. Einsetzen der stumpfen Nadel in den Samenbehälter des Hummers, um eine Spermaprobe zu extrahieren. Der Winkel und die Tiefe der Nadel beide kritisch, um konsistente Proben. Rechts: Blick in eine Samenbehälter, die seziert hat und vertikal teilen. Die Spermien Stecker belegt den oberen oder äußeren Abschnitt des Behälters, während die Spermien am Boden (tiefste Stelle) des Behälters befindet. Es ist wichtig zu beachten, dass die Abtastung unterhalb dieser Tiefe kann das Gefäß platzen und führen zu einer Blut kontaminierte Probe. <a href="https://www.jove.com/files/ftp_upload/50498/50498fig1highres.jpg" target="_blank">Klicken Sie hier für eine größere Ansicht</a> . 3. Eindringen in die Sperma-Stecker <ol><li> Nach der ersten Insertion, steiler der Winkel der Nadel fast senkrecht (senkrecht zur Körperachse) und sehr langsam beginnen, es zu arbeitennach unten in den Behälter. </li><li> Halten der Nadel in einem Winkel geneigt, so daß die Spitze leicht in Richtung des Schwanzes hingewiesen. In diesem Fall wird die Nadelspitze Komprimieren der Oberseite des Steckers, bis sie schließlich in die Matrix eindringt. </li><li> Fahren Sie langsam schieben die Nadel nach unten, bis ein Widerstand noch einmal gefühlt - das ist der Boden des Behälters und wo die Probe entnommen wird. </li></ol> 4. Sperma-Entfernung <ol><li> Entfernen einer Probe (oft feste Verschlussmaterial, flüssiges Sperma und andere Flüssigkeit) aus dem Boden des Behälters und die Hinterlegung der Probe in ein beschriftetes 2,0 ml Kunststoffrohr. </li><li> Spülen Sie den Behälter mit ~ 0,1-0,5 ml kaltem Meerwasser. </li><li> Im gleichen Röhrchen zu entfernen ungefähr 0,3 ml Flüssigkeit und Ort. </li><li> Shop Spermaproben auf Eis, bis sie im Labor untersucht werden. Teil B: Untersuchung von Spermaproben im Labor HINWEIS: Die Proben aus den Samen Aufnahmen der einzelnen l entfernt<html> obsters zu prüfen, um festzustellen, ob Spermien vorhanden oder nicht vorhanden waren werden. </li><li> Entfernen ~ 50 ul von Flüssigkeit aus jedem Kunststoffrohr, auf ein Glasobjektträger mit einem Deckglas und Blick auf 100X mit einer Verbindung Mikroskop. </li><li> Note Proben als: Sperma knapp, zahlreiche, oder nicht vorhanden (diese Studie nicht quantifizieren tatsächlichen Spermienzahlen, Abbildung 2). </li></ol><img alt="Figur 2" fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig2highres.jpg" src="/files/ftp_upload/50498/50498fig2.jpg" width="600px" /> 2. Lobster Spermaprobe Bilder. Bilder von Hummer Spermaproben aus Samen Aufnahme einer Frau getroffen, wie mit einer Verbindung Mikroskop bei 40-facher Vergrößerung betrachtet. Bild links zeigt ein Beispiel, die zahlreiche Samen enthalten, während die andere (rechts) zeigt eine spärlich Probe (Maßstab = 10 um).. Jpg &quot;target =&quot; _blank &quot;&gt; Klicken Sie hier für eine größere Ansicht.

<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>

Die Autoren erklären, keine Interessenkonflikte.

Dieses Video-und Papier Umriss und zeigen ein Verfahren zur Feststellung, ob weibliche Hummer erfolgreich gepaart. Dieser Ansatz ist einfach genug, daß sie für große Probennahme von Hummern auf See durch eine Vielzahl von Benutzern verwendet werden. Die Methode basiert auf dem Nachweis der Anwesenheit einer Spermatophore in der Samenbehälter, die von männlichen zu weiblichen Hummer während der Paarung übergeben wird, basiert. Diese Spermatophoren teilweise in einer Samen Stecker härten, kurz nachdem sie in den B...

<html> Amerikanische Hummer (Homarus americanus) ist einer der produktivsten Fischerei im Nordatlantik (~ 56.000 mt im Jahr 2011 über 390 Millionen US-Dollar) ein. Allerdings gibt es einen allgemeinen Mangel an Verständnis über die Fortpflanzungsdynamik dieser Art in Wildpopulationen. Generieren genauere Schätzungen der Reproduktionsleistung, einschließlich der Anzahl und Größen von Einzelpersonen aktiv in der Reproduktion beteiligt, könnte die Bestandsabschätzung zu verbessern. Langusten 2, 3 blaue Krabben, Königskrabben 4, Steinkrabben 5, Krabben und Schnee: Zum Beispiel haben Frauen, die von der Erreichung ihrer vollen Reproduktionspotenzial durch Spermien Begrenzung verhindert werden als ein Problem für einige kommerziell genutzten marinen Krebstieren einschließlich identifiziert 6. Das übergeordnete Ziel ist es, festzustellen, ob Spermien Einschränkung könnte ein Faktor in bestimmten Regionen der amerikanische Hummer-Fischerei als gut. nt &quot;&gt; Die ersten Anzeichen einer möglichen Einschränkung Spermien Problem in Hummer wurden beobachtet, während der Durchführung einer unabhängigen Tagging-Studie mit ovigerous weibliche Hummer (mit sichtbar intakt, vor kurzem extrudiert Eier). Eroberung Berichte von Fischer darauf hin, dass etwa 15% dieser Tiere hatte fallen lassen ihre Eigelege nach nur 1-2 Monate. Die Arbeitshypothese war, dass einige Hummer trugen Eier, die nicht befruchtet worden war, und als Ergebnis, &quot;fiel&quot; 7. Vorläufige Daten aus einer Folgestudie haben die Tatsache bestätigt, dass Hummer wird extrudieren, Eier, auch wenn sie nicht erfolgreich gepaart und diese unbefruchteten Eiern werden nur für ~ 1 Monat 8 durchgeführt. daher, angesichts der Beobachtung der Durchführung unbefruchteten Eiern Weibchen in natürlichen Populationen haben wir versucht, das Ausmaß, in dem Sperma Einschränkung vielleicht beitragen bestimmen eine submaximale Niveau der Wiedergabe durch geschlechtsreif Hummer. Um dieses Ziel zu erreichen, wurde eine Technik entwickelt, um die spermato erkennenphore während der Paarung vom Männchen hinterlegt. Dieses Papier-und Video beschreiben eine zur Ermittlung der Paarungserfolg der weiblichen Hummer entwickelt nicht-invasive, einfache Methode. Die Technik kann schnell und zuverlässig auf dem Meer, an Bord entweder Forschung oder kommerzielle Fischereifahrzeugen genutzt werden. Die Einzelheiten dieses Abtastverfahren sowie einige repräsentative Ergebnisse werden dargestellt, um die Anwendung der Technik veranschaulichen.

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

Obwohl er einer der produktivsten Fischerei im Nordwestatlantik, bleibt noch viel Unbekanntes über den natürlichen Fortpflanzungsdynamik der amerikanischen Hummer. Neuere Arbeiten genutzt Krustentier-Populationen (Krabben und Hummer) legt nahe, dass es Umstände gibt, wo reife Frauen sind nicht in der Lage, ihre volle Reproduktionspotenzial durch Spermien Begrenzung erreichen. Um diese Möglichkeit in verschiedenen Regionen der amerikanischen Hummer Fischerei zu untersuchen, wurde ein zuverlässiges und nichtinvasive Verfahren zum Abtasten einer großen Anzahl von weiblichen Hummer auf See entwickelt. Dieses Verfahren beinhaltet das Einführen einer stumpfen Kanüle in Samen Aufnahme der Frau, um die Anwesenheit oder Abwesenheit eines Steckers Spermien zu bestimmen und eine Probe, die auf das Vorhandensein von Spermien untersucht werden können, zu widerrufen. Eine Reihe von Kontroll-Studien wurden an der Anlegestelle und im Labor durchgeführt, um die Zuverlässigkeit dieser Technik testen. Diese Anstrengungen verbundenen Stichproben 294 weibliche Hummer, um zu bestätigen, dass die Anwesenheit ofa Spermien Stecker war ein verlässlicher Indikator für die Spermien innerhalb der Aufnahme und damit der Paarung. Dieses Papier beschreibt die Methodik und die aus einer Teilmenge der gesamten Stichprobe Frauen erzielten Ergebnisse. Von den 230 weiblichen Hummer aus George Bank und Cape Ann, MA abgetastet (Größenbereich von 71 bis 145 mm = Panzerlänge), waren 90,3% positiv für Spermien. Mögliche Erklärungen für das Fehlen von Spermien in einigen Weibchen sind: Unreife (mangelnde physiologische Reife), Ausfall der Spermien nach dem Stecker verwendet, um ein Gelege zu befruchten, und der Mangel an passenden Aktivität. Die Umfragen zeigen, dass diese Technik für die Untersuchung der Paarungserfolg der weiblichen Hummer ist ein zuverlässiger Proxy, der auf dem Gebiet verwendet werden, um Fortpflanzungsaktivität in natürlichen Populationen dokumentiert werden kann.

<html> Die hier angegebenen Daten wurden für eine Teilmenge der weiblichen Hummer, die von beiden Near-und Offshore-Standorten entstanden. Die Nearshore-Gruppe bestand aus 44 Hummer, die in den Küstengewässern in der Nähe von Cape Ann, Massachusetts gefangen genommen und in Kühlboxen ins Labor zur Untersuchung transportiert wurden. Insgesamt 186 Offshore-Hummer wurden auf George Bank gefangen genommen und in einem kommerziellen Betrieb Anlage in New Hampshire abgetastet. Spermaproben wur...

Watch this Scientific Journal Video about A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus at JoVE.com

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

Fischerei-induzierten Veränderungen zu nutzen Krustentier Fischerei, wie das amerikanische Hummer-Fischerei, könnte möglicherweise ihre Reproduktionsdynamik beeinflussen, was zu einer Verringerung der Paarungserfolg. Ziel der Studie war es, eine nichtinvasive Methode zur Ermittlung der Paarungserfolg der weiblichen Hummer, die physiologisch oder funktional ausgereift sein kann, zu entwickeln.

Eine nichtinvasive Methode zur<em&gt; In situ</em&gt; Bestimmung des Gegen Erfolg im Frauen amerikanischen Hummer (<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 Aufrufe.  University of New Hampshire.  Fischerei-induzierten Veränderungen zu nutzen Krustentier Fischerei, wie das amerikanische Hummer-Fischerei, könnte möglicherweise ihre Reproduktionsdynamik beeinflussen, was zu einer Verringerung der Paarungserfolg. Ziel der Studie war es, eine nichtinvasive Methode zur Ermittlung der Paarungserfolg der weiblichen Hummer, die physiologisch oder funktional ausgereift sein kann, zu entwickeln. 

Serie: 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.

Eine Technik, um Amerikanische Buche für Widerstand gegen die Buche Schildlaus (Bildschirm<em&gt; Cryptococcus fagisuga</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 ...

Forschung

Umwelt

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 eine verbesserte Inex Methode für<em&gt; In Situ</em&gt; Messung von partikulären und gelösten Verbindungen, die durch Active Suspension Feeders Verarbeitet

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.

Optimierte Verfahren zur Bestimmung der Adsorption von Phosphonaten auf Granular Ferric Hydroxid mit einer miniaturisierten Phosphor Entschlossenheit Methode

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 Hybridisierung Techniken für Paraffin-eingebetteten Erwachsenen Korallen Proben

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.

Ein Loop-vermittelten isothermen Verstärkung (Lampe) Assay für schnelle Identifizierung von Tabak 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.

Implantation eines neuen Mikro akustische Tags in Juvenile Pazifik Neunaugen und amerikanischer Aal

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&amp;#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.

Eine telemetrische, gravimetrische Plattform für die physiologische Phänotypisierung von Pflanzen-Umwelt-Interaktionen in Echtzeit

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.

Misteleradikator - Ein neuartiges Werkzeug zur gleichzeitigen mechanischen und chemischen Bekämpfung der Mistel

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

A Complete Method for Evaluating the Performance of Photocatalysts for the Degradation of Antibiotics in Environmental Remediation

Various antibiotics such as tetracycline, aureomycin, amoxicillin, and levofloxacin are found in large quantities in groundwater and soil systems, potentially leading to the development of resistant and multi-drug resistant bacteria, posing a threat to humans, animals, and environmental systems. Photocatalytic technology has attracted keen interest due to its rapid and stable treatment and direct use of solar energy. However, most studies evaluating the performance of semiconductor catalysts for the photocatalytic degradation of organic pollutants in water are currently incomplete. In this paper, a complete experimental protocol is designed to comprehensively evaluate the photocatalytic performance of semiconductor catalysts. Herein, rhombic dodecahedral silver phosphate was prepared by a simple solvent phase synthesis method at room temperature and atmospheric pressure. BrSubphthalocyanine/Ag3PO4 heterojunction materials were prepared by the solvothermal method. The catalytic performance of as-prepared materials for the degradation of tetracycline was evaluated by studying different influencing factors such as catalyst dosage, temperature, pH, and anions at atmospheric pressure using a 300 W xenon lamp as a simulated solar light source and a light intensity of 350 mW/cm2. Compared with the first cycle, the constructed BrSubphthalocyanine/Ag3PO4 maintained 82.0% of the original photocatalytic activity after five photocatalytic cycles, while the pristine Ag3PO4 maintained only 28.6%. The stability of silver phosphate samples was further tested by a five-cycle experiment. This paper provides a complete process for evaluating the catalytic performance of semiconductor catalysts in the laboratory for the development of semiconductor catalysts with potential for practical applications.

Presented here is a protocol to explore a universal set of experimental procedures for comprehensive laboratory evaluation of photocatalysts in the field of environmental purification, using the example of photocatalytic removal of antibiotic organic pollutant molecules from water by phthalocyanine sensitized silver phosphate composites.

Eine vollständige Methode zur Bewertung der Leistung von Photokatalysatoren für den Abbau von Antibiotika in der Umweltsanierung

Various antibiotics such as tetracycline, aureomycin, amoxicillin, and levofloxacin are found in large quantities in groundwater and soil systems, ...

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 Bodenfeuchtesensoren in ungestörten Böden

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