Gli autori desiderano ringraziare tutti i pescatori di aragoste commerciali del New England cui le barche ci hanno permesso di campionare - la loro cooperazione è molto apprezzato. Inoltre, ringraziamo Little Bay Lobster Company (Newington, NH) e di Champlin Seafood (Point Judith, RI), che ci ha permesso di condurre il campionamento presso le loro strutture. I seguenti studenti UNH fornito la preziosa assistenza nel corso di questo studio: Haley Bianco, Françoise Morrison, Sarah Havener, Audra Chaput, e maggio Grose. Questo progetto è stato sostenuto da sovvenzioni dal NH SeaGrant a WHW (progetto # R/CFR-11) e un programma UNH Marine Grant JSG.

Parte A: Tecnica Campo di campionamento 1. Femmina Misure <ol><li> Per ogni aragoste femmine, misurare la lunghezza carapace (CL) e la larghezza del secondo segmento addominale con l&#39;approssimazione di 1,0 millimetri usando un paio di pinze. NOTA: Il secondo segmento addominale è misurata perché la larghezza dell&#39;addome (dove femmine portano le loro uova) è un indicatore della maturità sessuale. Inoltre, potrebbe essere vantaggioso per accertare la fase muta per abbinare successivamente questi dati con fase riproduttiva (vedere fase 1.2). </li><li> Ritagliare una piccola porzione distale fuori una delle pleopodi addominali (un protocollo metodologico standard aragoste) e memorizzare il pleopod in acqua di mare pulita per una visione successiva con un microscopio dissezione 9. NOTA: Questo passaggio potrebbe anche rivelare la presenza di ghiandole di cemento sui pleopodi, che sono un&#39;indicazione che le donne si stanno preparando per estrudere le loro uova 10. </li></ol> 2. Needle Inseincidenza sulle <ol><li> Inserire l&#39;ago della siringa (18 G, vedi Tabella 1) ~ 1 centimetro nel ricettacolo seminale, angolati verso la coda a ~ 45 ° (Figura 1). NOTA: In aragoste con tappi spermatozoi intatti l&#39;ago potrebbe incontrare qualche resistenza durante l&#39;inserimento, quanto è spinto attraverso il tappo di sperma prima di raggiungere la massima concentrazione di sperma sul fondo del recipiente seminale (Figura 1). NOTA: La presenza della spina, di per sé, è un indicatore affidabile che le aragoste sono accoppiati. In alcuni animali ci può essere poca resistenza, forse perché non spermatoforo è presente, il spermatoforo è piccola, non è ancora indurito in una spina, o potrebbe essere già stato utilizzato per concimare uno o più grinfie di uova. </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. Inserimento dell&#39;ago e la vista di aragosta presa seminale. Inserimento dell&#39;ago a punta smussata nella presa seminale dell&#39;aragosta per estrarre un campione di sperma. L&#39;angolo e la profondità dell&#39;ago sono entrambi fondamentali per ottenere campioni omogenei. A destra: Veduta di un ricettacolo seminale che è stato sezionato e diviso verticalmente. La spina sperma occupa la parte superiore o esterna del recipiente mentre gli spermatozoi si trovano in basso (punto più profondo) del recipiente. E &#39;importante notare che il campionamento di sotto di questa profondità può forare la presa e il risultato in un campione di sangue contaminato. <a href="https://www.jove.com/files/ftp_upload/50498/50498fig1highres.jpg" target="_blank">Clicca qui per vedere l&#39;immagine ingrandita</a> . 3. Penetrando la Sperm Plug <ol><li> Dopo l&#39;inserimento iniziale, steepen l&#39;angolo del dell&#39;ago pressoché verticali (perpendicolari all&#39;asse del corpo), e molto lentamente inizia a lavorareverso il basso nel recipiente. </li><li> Tenere l&#39;ago inclinato a un angolo in modo che la punta è rivolta leggermente verso la coda. In questo caso la punta dell&#39;ago si comprime la parte superiore del tappo, finché alla fine penetra nella matrice. </li><li> Continuare a spingere lentamente l&#39;ago verso il basso fino a sentire resistenza una volta di più - questo è il fondo del recipiente e dove viene prelevato il campione. </li></ol> 4. Sperm rimozione <ol><li> Rimuovere un campione (spesso materiale spina solido, sperma liquido e di altro fluido) dal fondo del recipiente e depositare il campione in una provetta di plastica etichettata 2.0 ml. </li><li> Lavare il recipiente con ~ 0,1-0,5 ml di acqua di mare fredda. </li><li> Rimuovere circa 0,3 ml di liquido e di luogo nella stessa provetta. </li><li> Conservare i campioni di sperma sul ghiaccio fino a che non possono essere esaminati in laboratorio. Parte B: Esame dei campioni di sperma in laboratorio NOTA: I campioni prelevati dai recipienti seminali dei singoli l<html> obsters devono essere esaminati per determinare se gli spermatozoi erano presenti o assenti. </li><li> Rimuovere ~ 50 ml di liquido da ogni tubo di plastica, posto su un vetrino con una scivolata di copertura, e vista a 100X con un microscopio composto. </li><li> Punteggio campioni: lo sperma scarsa, numerosi, o assente (questo studio non ha quantificato il numero di spermatozoi effettivi, 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. Immagini campione Lobster sperma. Immagini di campioni di aragosta sperma prelevati da ricettacolo seminale di una femmina, come visto con un microscopio composto a 40X di ingrandimento. Immagine sulla sinistra mostra un esempio che conteneva numerosi sperma mentre l&#39;altro (destra) illustra un campione sparse (bar scala = 10 um).. Jpg &quot;target =&quot; _blank &quot;&gt; Clicca qui per vedere l&#39;immagine ingrandita.

<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>

Gli autori dichiarano assenza di conflitti di interesse.

Questo schema video e carta e dimostrare un metodo per determinare se le aragoste femmine sono accoppiati con successo. Questo approccio è abbastanza semplice che può essere utilizzato per il campionamento larga scala di aragoste in mare da una varietà di utenti. Il metodo si basa sulla rilevazione della presenza di un spermatophore nel ricettacolo seminale che viene passato da maschio a aragoste femmina durante l&#39;accoppiamento. Questi spermatofori parzialmente induriscono in una spina spermatozoi poco dopo che s...

<html> Astice americano (Homarus americanus) è una delle attività di pesca più produttive del Nord Atlantico (~ 56.000 tonnellate nel 2011, valutato più di 390 milioni dollari) 1. Tuttavia, vi è una generale mancanza di comprensione riguardo le dinamiche riproduttive di questa specie in popolazioni selvatiche. Generazione di stime più accurate di potenza riproduttiva, compreso il numero e le dimensioni delle persone che partecipano attivamente alla riproduzione, può migliorare il processo di valutazione degli stock. Ad esempio, le femmine che sono impedito di raggiungere il loro pieno potenziale riproduttivo a causa delle limitazioni spermatozoi sono stati identificati come una preoccupazione per alcuni crostacei marini sfruttate commercialmente, tra cui: aragoste, granchi blu 2 3, re granchi 4, 5 granchi di pietra, e granchi neve 6. L&#39;obiettivo generale è quello di stabilire se la limitazione dello sperma potrebbe essere un fattore in alcune regioni del aragosta pesca americano. nt &quot;&gt; I primi segni di un potenziale problema di limitazione di spermatozoi nel aragoste sono stati osservati durante lo svolgimento di uno studio di tagging correlato con ovigeri aragoste femmine (con visibilmente intatti, uova di recente estrusi). Recapture relazioni di pescatori ha indicato che circa il 15% di questi animali era caduto loro grinfie di uova dopo soli 1-2 mesi. L&#39;ipotesi di lavoro è che alcune aragoste portavano le uova che non erano state fecondate e, di conseguenza, &#39;caduto&#39; 7. dati preliminari di uno studio successivo hanno confermato il fatto che le aragoste saranno estrudere uova, anche se non sono accoppiati con successo e queste uova non fecondate vengono effettuate solo per ~ 1 mesi 8. Pertanto, data l&#39;osservazione delle femmine che trasportano le uova non fecondate nelle popolazioni naturali, abbiamo cercato di determinare la misura in cui la limitazione dello sperma potrebbe essere contribuendo a un livello submassimale di riproduzione di aragoste sessualmente maturi. Per raggiungere questo obiettivo, una tecnica è stata sviluppata per rilevare la spermatozoiPhore depositato dai maschi durante l&#39;accoppiamento. Questo documento e il video descrivono un non invasivo, semplice metodo sviluppato per accertare il successo di accoppiamento di aragoste femmine. La tecnica può essere utilizzata in modo rapido e affidabile al mare, sia a bordo delle ricerche o pescherecci commerciali. I dettagli del metodo di campionamento nonché alcuni risultati rappresentativi sono presentati per illustrare l&#39;applicazione della tecnica.

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			<td height="40" style="height:40px;width:163px;">Calipers: Mitutoyo Digimatic</td>
			<td style="width:129px;">Mitutoyo</td>
			<td style="width:135px;">500-196-20</td>
			<td style="width:231px;">http://www.globalindustrial.com</td>
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			<td height="40" style="height:40px;width:163px;">3 mL Luer-Lok&#8482; syringe</td>
			<td style="width:129px;">Beckton-Dickinson</td>
			<td style="width:135px;">309585</td>
			<td style="width:231px;">www.bd.com</td>
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			<td height="10" style="height:10px;width:163px;"></td>
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			<td height="60" style="height:60px;width:163px;">Monoject aluminum hub blunt needles, 18-G x 1</td>
			<td style="width:129px;">Webster Veterinary</td>
			<td style="width:135px;">8881202348</td>
			<td style="width:231px;">www.mywebstervet.com</td>
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			<td height="40" style="height:40px;width:163px;">2.0 mL plastic storage tubes</td>
			<td style="width:129px;">Eppendorf&#160;</td>
			<td style="width:135px;">wu-06333-72</td>
			<td style="width:231px;">http://www.coleparmer.com</td>
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			<td height="40" style="height:40px;width:163px;">Styrofoam cooler with ice</td>
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			<td height="40" style="height:40px;width:163px;">Compound light microscope</td>
			<td style="width:129px;">Olympus&#160;</td>
			<td style="width:135px;">BH System</td>
			<td style="width:231px;">http://www.olympusamerica.com</td>
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			<td height="40" style="height:40px;width:163px;">Glass sides with cover slip</td>
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			<td style="width:231px;">75 mm x 25 mm, ~ 1 mm thickness</td>
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			<td height="41" style="height:41px;width:163px;">Pipettor with disposable tips</td>
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a noninvasive method for in situ determination of mating success in female american lobsters (homarus americanus)

Nonostante sia una delle attività di pesca più produttive nell&#39;Atlantico nord-occidentale, molto rimane sconosciuto circa le dinamiche riproduttive naturali di aragoste americane. Un recente lavoro in popolazioni di crostacei sfruttate (granchi e aragoste), suggerisce che ci sono circostanze in cui le femmine mature sono in grado di raggiungere il loro pieno potenziale riproduttivo a causa delle limitazioni dello sperma. Per esaminare questa possibilità in diverse regioni del aragosta pesca americano, un metodo affidabile e non invasiva è stato sviluppato per il campionamento di un gran numero di aragoste femmine in mare. Questo metodo comporta l&#39;inserimento di un ago a punta smussata nel ricettacolo seminale della femmina per determinare la presenza o l&#39;assenza di una spina sperma e per prelevare un campione che può essere esaminato per la presenza di spermatozoi. Una serie di studi di controllo sono stati condotti al molo e in laboratorio per testare l&#39;affidabilità di questa tecnica. Questi sforzi hanno comportato campionamento 294 aragoste femmine per confermare che la presenza ospina sperma fa era un indicatore affidabile di sperma all&#39;interno del recipiente e, quindi, l&#39;accoppiamento. Dettagli Questo documento la metodologia ed i risultati ottenuti da un sottoinsieme delle femmine totale del campione. Dei 230 aragoste femmine nel campione da Bank George e Cape Ann, MA (range size = 71-145 mm di lunghezza carapace), il 90,3% era positivo per gli spermatozoi. Possibili spiegazioni per l&#39;assenza di spermatozoi in alcune femmine sono: immaturità (mancanza di maturità fisiologica), la ripartizione del plug sperma dopo essere stato utilizzato per fertilizzare una frizione di uova, e la mancanza di attività di accoppiamento. Le indagini indicano che questa tecnica per esaminare l&#39;accoppiamento di successo aragoste femmine è un proxy affidabile che può essere utilizzato nel campo per documentare attività riproduttiva in popolazioni naturali.

<html> I dati qui riportati sono stati per un sottoinsieme di aragoste femmine che provenivano sia da nearshore e sedi offshore. Il gruppo nearshore consisteva di 44 aragoste che sono stati catturati nelle acque costiere vicino a Cape Ann, Massachusetts e trasportate in dispositivi di raffreddamento al laboratorio per l&#39;esame. Un totale di 186 aragoste off-shore sono stati catturati sulla Banca di George e campionato in una struttura un&#39;azienda commerciale in New Hampshire. Campioni d...

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

Cambiamenti della pesca indotti alla pesca di crostacei sfruttati, come l&#39;aragosta pesca americano, potrebbero potenzialmente influenzare le loro dinamiche riproduttive, portando ad una riduzione nel successo di accoppiamento. L&#39;obiettivo di questo studio era di sviluppare un metodo invasivo per accertare il successo dell&#39;accoppiamento delle aragoste femmine che possono essere fisiologicamente o funzionalmente maturo.

Un metodo non invasivo per<em&gt; In situ</em&gt; Determinazione del successo di accoppiamento in americano femminile Aragoste (<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 Views.  University of New Hampshire.  Cambiamenti della pesca indotti alla pesca di crostacei sfruttati, come l'aragosta pesca americano, potrebbero potenzialmente influenzare le loro dinamiche riproduttive, portando ad una riduzione nel successo di accoppiamento. L'obiettivo di questo studio era di sviluppare un metodo invasivo per accertare il successo dell'accoppiamento delle aragoste femmine che possono essere fisiologicamente o funzionalmente maturo. 

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 tecnica a Screen Faggio americano per la Resistenza al Beech cocciniglia (<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 ...

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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 metodo migliorato per Inex<em&gt; In Situ</em&gt; Misura di particolato e sciolto composti elaborati da Alimentatori sospensioni attive

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.

Procedura ottimizzata per determinare l'adsorbimento di fosfonati su idrossido ferrico granulare utilizzando un metodo di determinazione del fosforo miniaturizzato

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 Tecniche di ibridazione per paraffina adulti coralli campioni

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'analisi di amplificazione isotermica (lampada) mediata da Loop per la rapida identificazione di 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.

Impianto di un nuovo Micro Tag acustico in Juvenile lampreda Pacifico e anguilla 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&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.

Una piattaforma telemetrica e gravimetrica per la fenotipizzazione fisiologica in tempo reale delle interazioni pianta-ambiente

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.

Mistletoe Eradicator - Un nuovo strumento per il controllo meccanico e chimico simultaneo del vischio

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.

Un metodo completo per valutare le prestazioni dei fotocatalizzatori per la degradazione degli antibiotici nella bonifica ambientale

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.

Sul posto Sensori di umidità del suolo in terreni indisturbati

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