Gli autori ringraziano il finanziamento del monitoraggio Forest Service Health Protection Evaluation US Forest e programmi di sviluppo della tecnologia speciale che hanno sostenuto lo sviluppo, la modifica e l&#39;applicazione del protocollo descritto qui di identificare gli alberi che sono resistenti e sensibili alla cocciniglia di faggio. Siamo anche grati al Holden Arboretum, Kirtland, OH, per il loro continuo collaborazione e il sostegno del nostro programma di screening resistenza faggio americano.

1 pianta Materiale:. Mature Campo Trees, in vaso piante, in vaso o innesti <ol><li> Per le prove sul campo, selezionare maturi sani faggi americani che mostrano segni di infestazione di scala o di malattia per i test di resistenza possibile. Visibilmente alberi sensibili dovranno anche essere identificati per essere usato come controllo (Figura 1). </li><li> Per testare piantine o innesti in vaso, raccogliere e germinare faggine come descritto nel Koch &amp; Carey, 2004 o rampollo innesto come descritto nella Carey et al, 2013. </li><li> Crescere piantine o innesti in invasatura miscela di suolo modificato con 47 g di micronutrienti, 477 g lenta concime 15N-3.9P-9.9K, 700 g perlite grossolana e 75 g di solfato di alluminio per 2.8 cu. borsa ft. Se necessario più avanti nella stagione di crescita, concimare le piante settimanali con solubile 17N-1.3P-14.1K a 200 ppm di azoto. </li><li> Durante la stagione di crescita, mantenere le piante in una casa all&#39;ombra. Permettono alle piante di andare dormiente fuori in autunno prima di spostarlidi un impianto di stoccaggio a temperatura controllata (~ 4 ° C), da novembre ad aprile. </li></ol> 2. Insieme di Faggio Scala uova <ol><li> In uno stand BBD infestata, ispezionare gli alberi pesantemente infestate (facilmente identificabili dalla loro &quot;imbiancato&quot; apparenza, figura 1A) con una lente di ingrandimento mano per confermare la presenza di uova, in genere abbondanti da metà luglio a metà agosto. </li><li> Utilizzare un pennello per spazzolare delicatamente le macchie cerose bianche di cocciniglie adulti, uova e altri detriti, in un sacchetto di plastica raccolta un gallone sigillabile (Figura 1B). Raccogliere da un minimo di tre alberi differenti almeno 12 m di distanza. </li><li> Se necessario, memorizzare uova nella sacca di raccolta sigillato per due settimane a 4 ° C. Un piccolo pezzo di nastro (2,5 cm quadrati) di inumidita polietilene espanso all&#39;interno del sacchetto per evitare che le uova si secchi. </li><li> Per separare le uova scala (0,15 x 0,25 mm) dagli adulti (0,60 millimetri) e detritis, costruire un setaccio con un corto pezzo di 2 &#39;tubo in PVC ed un giunto a sostenere un pezzo quadrato di 250 micron maglia di nylon (Figura 2). </li><li> Svuotare la miscela di adulti, cera, uova e detriti dalla sacca di raccolta sul setaccio e utilizzare un piccolo pennello per incoraggiare delicatamente le uova di passare attraverso le maglie in una piastra di Petri di vetro sotto. Piatti di plastica Petri dovrebbero essere evitati perché in possesso di elettricità più statica, rendendo difficile per spostare le uova. Uova prima e dopo setacciatura sono mostrati in Figura 3. </li><li> Uova purificati possono essere memorizzati nella capsula di Petri a 4 ° C per almeno una settimana. Per evitare che le uova si asciughi, nastro adesivo un pezzo umido di schiuma al coperchio e sigillare con Parafilm. </li></ol> 3. Egg Viabilità saggi <ol><li> Per valutare la vitalità uovo, utilizzare una siringa da 10 cc di applicare un sottile anello di vaselina intorno alla circonferenza della parte inferiore di una piastra di Petri 60 millimetri di vetro (Figura 4A </strong&gt;). </li><li> Trasferire circa 100 uova al centro del ring, mettere il coperchio sulla piastra di Petri e sigillare con Parafilm. Lasciare la piastra di Petri sigillata rimanga indisturbato a temperatura ambiente per 2 settimane o 3 settimane se uova sono stati conservati a 4 ° C prima di iniziare il test. </li><li> Ninfe tratteggiate saranno bloccati nella vaselina e possono essere facilmente contato, e le uova vuote sono facilmente distinguibili dalle uova non schiuse per il loro colore e la lucentezza (Figura 4B). Calcolo della redditività dividendo il numero di ninfe dalla somma delle uova vuoti più le rimanenti uova completi. Buona redditività dovrebbe essere nel 75% al ​​90% schiusi gamma uova. </li></ol> 4. Resistenza Scala Proiezione del Grande matura Faggi nel campo <ol><li> Per un test quantitativo, contare su 500 uova utilizzando un microscopio da dissezione e utilizzando una piccola spatola delicatamente cospargere attraverso il centro di un predampened, a celle aperte 10 x 15 x 1,3 centimetri rettangolo di polietilene espanso. Asmorzare schiuma, bagnarlo poi spremere quanta più acqua possibile. Per un test qualitativo, 500 uova possono essere contate e collocati in una piccola fiala di vetro e una linea di &quot;riempimento&quot; disegnata che può essere utilizzato per misurare ulteriori batch di circa 500 uova. </li><li> Posizionare il rilievo di gomma piuma sull&#39;albero prova con il lato rivolto verso uovo contro la corteccia. Tenere il pad in luogo con la corda, corda, spago o filo hardware plastificato. Materiale plastico o metallico a base dovrebbero essere utilizzati invece di materiali in fibra naturali che sono più facilmente eliminato dalla fauna selvatica. </li><li> Linea la parte superiore e lati di un cm pezzo di pellicola permeabile al vapore casa impermeabile con base di acetato adesivo siliconico pretagliati 23 x 30 e posizionarlo sopra il pad test di schiuma. Premere i bordi della pellicola fino alla albero per creare una tenuta stagna. Mettere spago di nylon o filo hardware rivestito di plastica intorno all&#39;albero e avvolgere casa, per tenerlo in posizione mentre l&#39;adesivo (Figura 5). </li><li> Posizionare un minimo di duetamponi di test di ciascun albero di test, preferibilmente su lati opposti del bolo. In ogni sito, mettere tamponi di test su almeno 2 alberi sensibili (con evidente infestazione scala naturale) come controllo. Prima di posizionare gli elettrodi di prova su alberi di comando, rimuovere le cocciniglie naturali o uova con una ditta di setole pennello. Nota: pastiglie schiuma con le uova possono anche essere collocati sugli alberi al fine di uova scala allevamento, che possono essere particolarmente utili nelle zone dove il livello di infestazione è basso. </li></ol> 5. Resistenza Scala Proiezione del vaso Piantine o innesti <ol><li> Per il test, selezionare alberi in vaso con un diametro minimo di 1 cm (pinza a 5 cm sopra la linea del suolo) che sono abbastanza alto per avere almeno 2 pastiglie di test separati affidamento su di esse. Potare piccoli rami laterali, se necessario, per fare spazio per le pastiglie di prova. </li><li> Utilizzare un microscopio dissezione a contare 150 uova e spargere sopra un pretagliati e inumidito 2,5 x 7,6 x 1,3 centimetri polietilene a cellule aperterilievo di gomma piuma lene. </li><li> Fissare il pad schiuma piantina con il lato uovo contro la corteccia, con filo plastificato. Avvolgere un piccolo quadrato di avvolgere casa intorno alla piantina appena sopra la schiuma e sigillare con silicone acetato-based (vedere Figura 6A). </li><li> Riportare le famiglie di piantine sensibili o innesti come controlli. </li></ol> 6. Raccolta dati Nota: circa 52-57 settimane dopo l&#39;immissione le spugne e le uova sugli alberi di prova, i dati possono essere raccolti. E &#39;importante che questo non è fatto solo dopo adulti hanno cominciato a deporre le uova in modo che la loro capacità di riprodursi può essere determinato. Su alcuni alberi resistenti non è raro vedere un piccolo numero di adulti stabilire ma si verificano nessuna riproduzione. <ol><li> Rimuovere con attenzione il rilievo di gomma piuma e contare il numero di cocciniglie adulto stabiliti sulla corteccia utilizzando una lente o lente di ingrandimento (10X). </li><li> Quando il tampone di schiuma viene rimosso, non è uNcommon per alcuni adulti e la maggior parte dei cluster di uova di essere tirato fuori della struttura con la schiuma (vedi Figura 7). Utilizzando un microscopio da dissezione, contare i cluster di uova e gli adulti che sono rimasti attaccato al tappetino in gomma. </li></ol>

<ol>
	<li>Houston, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+to+artificially+infest+beech+bark+with+the+scale,+Cryptococcus+fagisuga+(Lindinger).">A technique to artificially infest beech bark with the scale, Cryptococcus fagisuga (Lindinger).</a> U.S. Forest Service. , (1982).</li><li>Houston, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=American+beech+resistance+to+Cryptococcus+fagisuga.">American beech resistance to Cryptococcus fagisuga.</a> Proceeding, IUFRO beech bark disease working party conference. , 38-42 (1983).</li><li>Ehrlich, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+beech+bark+disease:+a+nectria+disease+of+Fagus,+following+Cryptococcus+fagi.">The beech bark disease: a nectria disease of Fagus, following Cryptococcus fagi.</a> 10, 593-692 (1934).</li><li>Papaik, M. J., Canham, C. D., Latty, E. F., Woods, K. D. <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+an+introduced+pathogen+on+resistance+to+natural+disturbance:+beech+bark+disease+and+withdrow.">Effects of an introduced pathogen on resistance to natural disturbance: beech bark disease and withdrow.</a> 35, 1832-1843 (2005).</li><li>Miller-Weeks, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+status+of+beech+bark+disease+in+New+England+and+New+York.">Current status of beech bark disease in New England and New York.</a> Proceeding, IUFRO beech bark disease working party conference. , 21-23 (1983).</li><li>Houston, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beech+bark+disease:+The+aftermath+forests+are+structured+for+a+new+outbreak.">Beech bark disease: The aftermath forests are structured for a new outbreak.</a> J. For. 73, 660-663 (1975).</li><li>Twery, M. J., Patterson, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variations+in+beech+bark+disease+and+its+effects+on+species+composition+and+structure+of+northern+hardwood+stands+in+central+New.">Variations in beech bark disease and its effects on species composition and structure of northern hardwood stands in central New.</a> 14, 565-574 (1984).</li><li>Jakubus, W. J., McLaughlin, C. R., Jensen, P. G., McNulty, S. A., Evans, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alternate+year+beechnut+production+and+its+influence+on+bear+and+marten+populations.">Alternate year beechnut production and its influence on bear and marten populations.</a> Beech Bark Disease: Proceedings of the Beech Bark Disease Symposium. , 79-87 (2005).</li><li>Koch, J. L., Carey, D. W., Mason, M. E., Nelson, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+beech+scale+resistance+in+full-+and+half-sib+families.">Assessment of beech scale resistance in full- and half-sib families.</a> Can. J. For. Res. 40, 265-272 (2010).</li><li>Koch, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beech+bark+disease:+The+oldest+&#8220;new&#8221;+threat+to+American+beech+in+the+United+States.">Beech bark disease: The oldest &#8220;new&#8221; threat to American beech in the United States.</a> Outlooks Pest Manag. 21, 64-68 (2010).</li><li>Koch, J. L., Mason, M. E., Carey, D. W., Palmieri, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Screening+for+resistance+to+beech+bark+disease:+improvements+and+results+from+seedlings+and+grafted+field+selections.">Screening for resistance to beech bark disease: improvements and results from seedlings and grafted field selections.</a> Proceedings of the 4th International Workshop on Genetics of Host-Parasite Interactions in Forestry. , (2012).</li><li>Leak, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fifty+year+impacts+of+the+beech+bark+disease+in+the+Bartlett+Experimental+Forest,+New+Hampshire.">Fifty year impacts of the beech bark disease in the Bartlett Experimental Forest, New Hampshire.</a> Northern J. Appl. For. 23, 141-143 (2006).</li><li>Koch, J. L., Carey, D. W., Yaussy, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+cross-pollinations+with+American+beech+trees+that+are+resistant+to+beech+bark+disease.">Controlled cross-pollinations with American beech trees that are resistant to beech bark disease.</a> Proceedings of the 14th Central Hardwood Forest Conference. , 358-364 (2004).</li><li>Carey, D. W., Mason, M. E., Bloese, P., Koch, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hot+callusing+for+propagation+of+American+beech+by+grafting.">Hot callusing for propagation of American beech by grafting.</a> HortSci. 48, 620-624 (2013).</li><li>Koch, J. L., Carey, W. W., Evans, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+resistance+of+American+beech+to+beech+bark+disease:+knowledge+through+2004.">The genetics of resistance of American beech to beech bark disease: knowledge through 2004.</a> Beech Bark Disease: Proceedings of the Beech Bark Disease Symposium. , 98-105 (2005).</li></ol>

Gli autori dichiarano di non avere interessi finanziari in competizione.

I passaggi critici necessari per il successo di questo test includono l&#39;esecuzione di un test di vitalità delle uova e l&#39;impiego di controlli sensibili in ogni sito testato, e con materiali vaso. E &#39;anche importante utilizzare più di un tampone di test per albero o piantina. Abbiamo trovato che sia nel campo e nelle prove più controllati su piante in vaso, una frequente fonte di errore può essere guasto pad. Ad esempio, nel campo, anche con nostra modifica di coprire il tampone di test con l&#39;involucr...

Beech malattia corteccia (BBD) ha avuto un impatto negativo sulla faggio americano in Nord America dopo l&#39;introduzione della cocciniglia faggio invasiva, Cryptococcus fagisuga, nella provincia canadese della Nuova Scozia alla fine del 1890 3. Questo complesso insetto-malattia è iniziata quando la cocciniglia faggio inserisce il suo stiletto di alimentazione nella corteccia creando piccole fessure che prevedono ingresso per l&#39;infezione da uno dei Neonectria specie di funghi (Neonectria ditissima o Neonectria faginata). Mentre i miceli fungini crescono, ampie aree di tessuto può morire, alla fine cingendo completamente l&#39;albero. Il danno dalla malattia indebolisce l&#39;albero, il che rende incline a rompersi in venti forti 4. Livelli di mortalità nella prima ondata della malattia sono stati segnalati per essere alto come il 50% 5. Alberi sopravvissuti sono spesso gravemente deformati come cancri formano ridurre il valore dell&#39;albero come un prodotto di legno.0; Tali alberi hanno una propensione per radice di germinazione che porta alla formazione di macchie &quot;faggio&quot; che impediscono altre specie più desiderabili introduca, riducendo il valore economico ed ecologico del supporto 6. Sebbene la malattia corteccia del faggio non rischia di portare all&#39;estinzione del faggio americano, altera stare composizione e la salute porta ad una diminuzione del cibo e habitat per la fauna selvatica 7,8. In stand affetti da BBD per molti anni, sono stati segnalati alberi che rimangono privi di qualsiasi sintomo della malattia. Prove di inoculazione artificiale hanno dimostrato che questi alberi sono resistenti all&#39;insetto scala 2. Senza infestazione scala, ci sono poche possibilità di infezione Neonectria, minimizzando l&#39;impatto del fungo. Mortalità larga scala in faggio americano a causa di infezione Neonectria in assenza di infestazione scala prima ha maistati segnalati, così resistenza alla scala faggio risultati insetti in resistenza BBD. Recenti ricerche sulla gestione dei BBD è concentrata sull&#39;identificazione, la propagazione, l&#39;allevamento, e il mantenimento di faggi americani con resistenza alla cocciniglia di faggio. Studi genetici hanno dimostrato che la resistenza alla cocciniglia è ereditabile e di un&#39;attenta selezione e l&#39;allevamento di alberi resistenti può portare a un significativo miglioramento in una sola generazione 9. Questa scoperta ha alimentato gli sforzi di stato e forestali nazionali manager negli Stati Uniti per stabilire arboreti da seme regionali di resistenti faggio americano per fornire una fonte di sementi geneticamente diverse BBD resistenti alle piantagioni di restauro 10,11. La ricerca ha anche indicato che la manipolazione selvicolturale della genetica dello stand dalla rimozione di alberi sensibili e conservazione di alberi resistenti può provocare in stand miglioramento 9,12. &lt;/ P&gt; Gestione delle BBD attraverso attività di miglioramento degli alberi o svolgendo prescrizioni selvicolturali richiede la capacità di selezionare e distinguere tra faggi scala resistente e suscettibile. I metodi qui presentati sono stati adattati da un metodo introdotto da Dave Houston per inoculare artificialmente piantine con le uova scala faggio 1. Il metodo può essere usato come strumento di screening per identificare quantitative trait loci (QTL) associati a resistenza o di distinguere tra piantine in vaso resistenti e suscettibili o ramet innestati negli studi genetici. In alternativa, può essere utilizzato per lo screening alberi maturi nel campo per identificare alberi resistenti allo sviluppo frutteto seme, o ritenzione nel campo. Alberi sensibili, possano essere identificati e rimossi per ridurre al minimo l&#39;impatto della malattia.

<table border="0" cellpadding="0" cellspacing="0" width="1006">
	<tbody>
		<tr height="42">
			<td height="63" rowspan="2" style="height:63px;width:163px;">Nylon Mesh Sheet, 250 micron</td>
			<td rowspan="2" style="width:120px;">Small Parts</td>
			<td rowspan="2" style="width:99px;">CMN-0250-D</td>
			<td style="width:627px;">Mesh opening size: 250 microns square, % of open area: 34, thread diameter: 180 micron, width: 24&#8221;, Length: 12&#8221;, Package quantity: 1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:627px;"><a href="http://www.amazonsupply.com/dp/B000FMWH1S/ref=sp_dp_g2c_asin">http://www.amazonsupply.com/dp/B000FMWH1S/ref=sp_dp_g2c_asin</a></td>
		</tr>
		<tr height="21">
			<td height="42" rowspan="2" style="height:42px;width:163px;">Tyvek Home Wrap</td>
			<td rowspan="2" style="width:120px;">DuPont</td>
			<td rowspan="2" style="width:99px;">D1349991</td>
			<td style="width:627px;">9 ft x 150 ft roll, can ask for it to be cut in half.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:627px;"><a href="http://www.lowes.com/pd_14582-2020-D13499991_0__">http://www.lowes.com/pd_14582-2020-D13499991_0__</a></td>
		</tr>
		<tr height="21">
			<td height="42" rowspan="2" style="height:42px;width:163px;">Polyethylene foam, &#189;&#8221; thick&#160;</td>
			<td rowspan="2" style="width:120px;">Columbus Foam Products</td>
			<td rowspan="2" style="width:99px;">N/A</td>
			<td style="width:627px;">2 ft x 50 ft roll</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:627px;"><a href="http://www.colsfoam.com/apcfp/products.htm">http://www.colsfoam.com/apcfp/products.htm</a></td>
		</tr>
		<tr height="42">
			<td height="64" rowspan="2" style="height:64px;width:163px;">MetroMix 510</td>
			<td style="width:120px;">Sun Gro&#160;</td>
			<td rowspan="2" style="width:99px;">N/A</td>
			<td style="width:627px;">2.8 cu. ft. bark, peat moss, vermiculite, bark ash, starter nutrient charge (with Gypsum), slow release nitrogen and&#160;dolomitic limestone.</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:120px;">Horticulture</td>
			<td style="width:627px;"><a href="http://sungro.com/products_displayProduct.php?product_id=118&amp;brand_id=17">http://sungro.com/products_displayProduct.php?product_id=118&#38;brand_id=17</a></td>
		</tr>
		<tr height="21">
			<td height="42" rowspan="2" style="height:42px;width:163px;">Osmocote Plus 15-9-12</td>
			<td rowspan="2" style="width:120px;">Everris</td>
			<td rowspan="2" style="width:99px;">E903206</td>
			<td style="width:627px;">Standard 3-4 month release.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:627px;"><a href="http://everris.us.com/product/15-9-12">http://everris.us.com/product/15-9-12</a></td>
		</tr>
		<tr height="21">
			<td height="42" rowspan="2" style="height:42px;width:163px;">Sight Savers 10 X</td>
			<td rowspan="2" style="width:120px;">Bausch &#38; Lomb</td>
			<td rowspan="2" style="width:99px;">813434</td>
			<td style="width:627px;">10 x magnification illuminated coddington</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:627px;"><a href="http://www.amleo.com/Bausch---Lomb-Illuminating-Codington-Magnifier-10x/p/CM10/">http://www.amleo.com/Bausch---Lomb-Illuminating-Codington-Magnifier-10x/p/CM10/</a></td>
		</tr>
		<tr height="21">
			<td height="42" rowspan="2" style="height:42px;width:163px;">Nikon Mini Field Stereoscope</td>
			<td rowspan="2" style="width:120px;">Nikon</td>
			<td rowspan="2" style="width:99px;">7314</td>
			<td style="width:627px;">20 x magnification</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:627px;"><a href="http://www.opticsplanet.com/nikon-fieldmicroscope.html">http://www.opticsplanet.com/nikon-fieldmicroscope.html</a></td>
		</tr>
		<tr height="21">
			<td height="42" rowspan="2" style="height:42px;width:163px;">Silicone II clear</td>
			<td rowspan="2" style="width:120px;">GE</td>
			<td rowspan="2" style="width:99px;">159538</td>
			<td style="width:627px;">2.9 oz clear window &#38; door caulk</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:627px;"><a href="http://www.midlandhardware.com/159538.html">http://www.midlandhardware.com/159538.html</a></td>
		</tr>
	</tbody>
</table>

a technique to screen american beech for resistance to the beech scale insect (cryptococcus fagisuga lind.)

Faggio corteccia malattia (BBD) si traduce in alti livelli di mortalità iniziale, lasciando dietro gli alberi superstiti che sono notevolmente indeboliti e deformati. La malattia è avviata da attività dell&#39;insetto scala faggio invasiva, Cryptococcus fagisuga, che crea punti di ingresso per l&#39;infezione da una delle specie di funghi Neonectria alimentazione. Senza infestazione scala, ci sono poche possibilità di infezione fungina. Utilizzo di uova scala per infestare artificialmente alberi sani in stand pesantemente colpiti BBD dimostrato che questi alberi erano resistenti alla porzione cocciniglia del complesso malattia 1. Qui vi presentiamo un protocollo che abbiamo sviluppato, basato sulla tecnica infestazione artificiale da Houston 2, che può essere usato per individuare alberi scala resistente nel settore e in piccole piantine in vaso e innesti. L&#39;identificazione degli alberi di scale-resistente è una componente importante della gestione del BBD attraverso alberoprogrammi di miglioramento e la manipolazione selvicolturale.

La Figura 6 mostra una piantina resistente (C) e due piantine sensibili (D, E) esibendo diversi gradi di sensibilità. Un esempio di ciò che un albero maturo suscettibile assomiglia a 57 settimane dopo la prova infestazione artificiale è stato istituito è mostrato in Figura 5B. Quando la schiuma viene staccato dall&#39;albero, non è raro per cocciniglie e loro grappoli d&#39;uovo per rimanere attaccato alla schiuma come mostrato in Figura 7,...

Watch this Scientific Journal Video about A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind. at JoVE.com

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

Beech malattia corteccia è iniziata dalle attività della cocciniglia di faggio che creano punti di ingresso fungine nella corteccia di alimentazione. Gli alberi che sono resistenti alla cocciniglia sono anche resistenti alle malattie. Qui vi presentiamo il protocollo che abbiamo sviluppato per lo screening individuali faggi per la resistenza scala di faggio.

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 ...

a-technique-to-screen-american-beech-for-resistance-to-beech-scale

Research

JoVE Journal

Environment

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

9.5K Views.  US Forest Service.  Beech malattia corteccia è iniziata dalle attività della cocciniglia di faggio che creano punti di ingresso fungine nella corteccia di alimentazione. Gli alberi che sono resistenti alla cocciniglia sono anche resistenti alle malattie. Qui vi presentiamo il protocollo che abbiamo sviluppato per lo screening individuali faggi per la resistenza scala di faggio. 

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

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

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American lobsters. Recent work in exploited crustacean populations (crabs and lobsters) suggests that there are circumstances where mature females are unable to achieve their full reproductive potential due to sperm limitation. To examine this possibility in different regions of the American lobster fishery, a reliable and noninvasive method was developed for sampling large numbers of female lobsters at sea. This method involves inserting a blunt-tipped needle into the female&#39;s seminal receptacle to determine the presence or absence of a sperm plug and to withdraw a sample that can be examined for the presence of sperm. A series of control studies were conducted at the dock and in the laboratory to test the reliability of this technique. These efforts entailed sampling 294 female lobsters to confirm that the presence of a sperm plug was a reliable indicator of sperm within the receptacle and thus, mating. This paper details the methodology and the results obtained from a subset of the total females sampled. Of the 230 female lobsters sampled from George&#39;s Bank and Cape Ann, MA (size range = 71-145 mm in carapace length), 90.3% were positive for sperm. Potential explanations for the absence of sperm in some females include: immaturity (lack of physiological maturity), breakdown of the sperm plug after being used to fertilize a clutch of eggs, and lack of mating activity. The surveys indicate that this technique for examining the mating success of female lobsters is a reliable proxy that can be used in the field to document reproductive activity in natural populations.

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

Fishery-induced changes to exploited crustacean fisheries, such as the American lobster fishery, could potentially influence their reproductive dynamics, leading to a reduction in mating success. This study&#39;s goal was to develop a noninvasive method for ascertaining the mating success of female lobsters that may be physiologically or functionally mature.

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 ...

Ricerca

Ambiente

Video Tracking Protocol to Screen Deterrent Chemistries for Honey Bees

The European honey bee, Apis mellifera L., is an economically and agriculturally important pollinator that generates billions of dollars annually. Honey bee colony numbers have been declining in the United States and many European countries since 1947. A number of factors play a role in this decline, including the unintentional exposure of honey bees to pesticides. The development of new methods and regulations are warranted to reduce pesticide exposures to these pollinators. One approach is the use of repellent chemistries that deter honey bees from a recently pesticide-treated crop. Here, we describe a protocol to discern the deterrence of honey bees exposed to select repellent chemistries. Honey bee foragers are collected and starved overnight in an incubator 15 h prior to testing. Individual honey bees are placed into Petri dishes that have either a sugar-agarose cube (control treatment) or sugar-agarose-compound cube (repellent treatment) placed into the middle of the dish. The Petri dish serves as the arena that is placed under a camera in a light box to record the honey bee locomotor activities using video tracking software. A total of 8 control and 8 repellent treatments were analyzed for a 10 min period with each treatment was duplicated with new honey bees. Here, we demonstrate that honey bees are deterred from the sugar-agarose cubes with a compound treatment whereas honey bees are attracted to the sugar-agarose cubes without an added compound.

The loss of honey bee colonies presents a challenge to crop pollination services. Current pollinator protection practices warrant an alternative approach to minimize the contact of honey bees to harmful pesticides using repellent chemistries. Here, we provide detailed methods for a visual tracking protocol to screen deterrents for bees.

Protocollo di monitoraggio video a chimici detergenti dello schermo per le api di miele

The European honey bee, Apis mellifera L., is an economically and agriculturally important pollinator that generates billions of dollars annually. Honey ...

Lab-Scale Model to Evaluate Odor and Gas Concentrations Emitted by Deep Bedded Pack Manure

A lab-scaled simulated bedded pack model was developed to study air quality and nutrient composition of deep-bedded packs used in cattle mono-slope facilities. This protocol has been used to effectively evaluate many different bedding materials, environmental variables (temperature, humidity), and potential mitigation treatments that can improve air quality in commercial deep-bedded mono-slope facilities. The model is dynamic and allows researchers to easily collect many chemical and physical measurements from the bedded pack. Weekly measurements, collected over the course of six to seven weeks, allows sufficient time to see changes in air quality measurements over time as the bedded pack matures. The data collected from the simulated bedded packs is within the range of concentrations previously measured in commercial deep-bedded mono-slope facilities. Past studies have demonstrated that 8 - 10 experimental units per treatment are sufficient to detect statistical differences among the simulated bedded packs. The bedded packs are easy to maintain, requiring less than 10 minutes of labor per bedded packs per week to add urine, feces, and bedding. Sample collection using the gas sampling system requires 20 - 30 minutes per bedded pack, depending on the measurements that are being collected. The use of lab-scaled bedded packs allows the researcher to control variables such as temperature, humidity, and bedding source that are difficult or impossible to control in a research or commercial facility. While not a perfect simulation of &#34;real-world&#34; conditions, the simulated bedded packs serve as a good model for researchers to use to examine treatment differences among bedded packs. Several lab-scale studies can be conducted to eliminate possible treatments before trying them in a research or commercial-sized facility.

A protocol has been developed to measure gases, odors, and nutrient composition in lab-scaled bedded manure packs, which can be used to study ways to improve air quality in commercial cattle facilities using deep-bedded manure packs.

Modello di scala di laboratorio per valutare odore e le concentrazioni di Gas emesse dal profondo Bedded Pack letame

A lab-scaled simulated bedded pack model was developed to study air quality and nutrient composition of deep-bedded packs used in cattle mono-slope ...

Double-stranded RNA Oral Delivery Methods to Induce RNA Interference in Phloem and Plant-sap-feeding Hemipteran Insects

Phloem and plant sap feeding insects invade the integrity of crops and fruits to retrieve nutrients, in the process damaging food crops. Hemipteran insects account for a number of economically substantial pests of plants that cause damage to crops by feeding on phloem sap. The brown marmorated stink bug (BMSB), Halyomorpha halys (Heteroptera: Pentatomidae) and the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Liviidae) are hemipteran insect pests introduced in North America, where they are an invasive agricultural pest of high-value specialty, row, and staple crops and citrus fruits, as well as a nuisance pest when they aggregate indoors. Insecticide resistance in many species has led to the development of alternate methods of pest management strategies. Double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) is a gene silencing mechanism for functional genomic studies that has potential applications as a tool for the management of insect pests. Exogenously synthesized dsRNA or small interfering RNA (siRNA) can trigger highly efficient gene silencing through the degradation of endogenous RNA, which is homologous to that presented. Effective and environmental use of RNAi as molecular biopesticides for biocontrol of hemipteran insects requires the in vivo delivery of dsRNAs through feeding. Here we demonstrate methods for delivery of dsRNA to insects: loading of dsRNA into green beans by immersion, and absorbing of gene-specific dsRNA with oral delivery through ingestion. We have also outlined non-transgenic plant delivery approaches using foliar sprays, root drench, trunk injections as well as clay granules, all of which may be essential for sustained release of dsRNA. Efficient delivery by orally ingested dsRNA was confirmed as an effective dosage to induce a significant decrease in expression of targeted genes, such as juvenile hormone acid O-methyltransferase (JHAMT) and vitellogenin (Vg). These innovative methods represent strategies for delivery of dsRNA to use in crop protection and overcome environmental challenges for pest management.

This article demonstrates novel techniques developed for oral delivery of double-stranded RNA (dsRNA) through the vascular tissues of plants for RNA interference (RNAi) in phloem sap feeding insects.

Metodi di consegna orale RNA double-stranded per indurre RNAi nel floema e insetti Pentatomomorfi pianta-sap-alimentazione

Phloem and plant sap feeding insects invade the integrity of crops and fruits to retrieve nutrients, in the process damaging food crops. Hemipteran ...

The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

Barnacles are marine crustaceans with a sessile adult and free-swimming, planktonic larvae. The barnacle Balanus (Amphibalanus) improvisus is particularly relevant as a model for the studies of osmoregulatory mechanisms because of its extreme tolerance to low salinity. It is also widely used as a model of settling biology, in particular in relation to antifouling research. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae for studies. A protocol for the all-year-round culturing of B. improvisus has been developed and a detailed description of all steps in the production line is outlined (i.e., the establishment of adult cultures on panels, the collection and rearing of barnacle larvae, and the administration of feed for adults and larvae). The description also provides guidance on troubleshooting and discusses critical parameters (e.g., the removal of contamination, the production of high-quality feed, the manpower needed, and the importance of high-quality seawater). Each batch from the culturing system maximally yields roughly 12,000 nauplii and can deliver four batches in a week, so up to almost 50,000 larvae per week can be produced. The method used to culture B. improvisus is, probably, to a large extent also applicable to other marine invertebrates with free-swimminglarvae. Protocols are presented for the dissection of various tissues from adults as well as the production of high-quality RNA for studies on gene expression. It is also described how cultured adults and reared cyprids can be utilized in a wide array of experimental designs for examining gene expression in relation to external factors. The use of cultured barnacles in gene expression is illustrated with studies of possible osmoregulatory roles of Na+/K+ ATPase and aquaporins.

The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

The barnacle Balanus (Amphibalanus) improvisus is a model for studying osmoregulation and antifouling. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae. Here, a protocol for the all-year-round culturing of B. improvisus is described, including the production of larvae. The use of cultured barnacles in gene expression studies is illustrated.

Il Barnacle Balanus BalanUs come modello Marine - coltura e l'espressione genica

Barnacles are marine crustaceans with a sessile adult and free-swimming, planktonic larvae. The barnacle Balanus (Amphibalanus) improvisus is particularly ...

Experimental Protocol for Examining Behavioral Response Profiles in Larval Fish: Application to the Neuro-stimulant Caffeine

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and toxicological studies. Using automated digital tracking platforms, recent efforts in neuropharmacology are leveraging larval fish locomotor behaviors to identify potential therapeutic targets for novel small molecules. Similar to these efforts, research in the environmental sciences and comparative pharmacology and toxicology is examining various behaviors of fish models as diagnostic tools in tiered evaluation of contaminants and real-time monitoring of surface waters for contaminant threats. Whereas the zebrafish is a popular larval fish model in the biomedical sciences, the fathead minnow is a common larval fish model in ecotoxicology. Unfortunately, fathead minnow larvae have received considerably less attention in behavioral studies. Here, we develop and demonstrate a behavioral profile protocol using caffeine as a model neurostimulant. Though photomotor responses of fathead minnows were occasionally affected by caffeine, zebrafish&#160;were markedly more sensitive for photomotor and locomotor endpoints, which responded at environmentally relevant levels. Future studies are needed to understand comparative behavioral sensitivity differences among fish with age and time of day, and to determine whether similar behavioral effects would occur in nature and be indicative of adverse outcomes at the individual or population levels of biological organization.

Here, we present a protocol to examine larval zebrafish and fathead minnow locomotor activities and photomotor responses (PMR) using an automated tracking software. When incorporated in common toxicology bioassays, analyses of these behaviors provide a diagnostic tool to examine chemical bioactivity. This protocol is described using caffeine, a model neurostimulant.

Protocollo sperimentale per l'esame di profili di risposta comportamentale nel pesce larvale: applicazione alla caffeina Neuro-stimolante

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and ...

A Precise and Autonomous System for the Detection of Insect Emergence Patterns

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of emerging insects they can detect. In order to obtain precise measurement of insect emergence, it is necessary for systems to be semi-automated and able to measure large numbers of emerging insects. We addressed these issues by designing and building a system that is automated and can measure emergence of up to 1200 insects. We modified the existing &#34;falling-ball&#34; system using Arduino microcontrollers to automate data collection and expand the sample size through multiple data channels. Multiple data channels enable the user to not only increase their sample size, but also allows for multiple treatments to be run simultaneously in a single experiment. Furthermore, we created an R script to automatically visualize the data as a bubble plot, while also calculating the median day and time of emergence. The current system was designed using 3D printing so the user can modify the system to be adjusted for different species of insects. The goal of this protocol is to investigate important questions in chronobiology and stress physiology, using this precise and automated system to measure insect emergence patterns.

Measurement of insect emergence patterns requires precision. Existing systems are only semi-automated and sample size is limited. We addressed these issues by designing a system using microcontrollers to precisely measure the time of emergence of large numbers of emerging insects.

Un sistema preciso ed autonomo per l'individuazione di modelli di emersione degli insetti

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

Composizione e analisi della distribuzione di Bioaerosols in diverse condizioni ambientali

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...

Detached Leaf Assays to Simplify Gene Expression Studies in Potato During Infestation by Chewing Insect Manduca sexta

The multitrophic nature of gene expression studies of insect herbivory demands large numbers of biological replicates, creating the need for simpler, more streamlined herbivory protocols. Perturbations of chewing insects are usually studied in whole plant systems. While this whole organism strategy is popular, it is not necessary if similar observations can be replicated in a single detached leaf. The assumption is that basic elements required for signal transduction are present within the leaf itself. In the case of early events in signal transduction, cells need only to receive the signal from the perturbation and transmit that signal to neighboring cells which are assayed for gene expression.
The proposed method simply changes the timing of the detachment. In whole plant experiments, larvae are confined to a single leaf which is eventually detached from the plant and assayed for gene expression. If the order of excision is reversed, from last in whole plant studies, to first in the detached study, the feeding experiment is simplified.
Solanum tuberosum var. Kennebec is propagated by nodal transfer in a simple tissue culture medium and transferred to soil for further growth if desired. Leaves are excised from the parent plant and relocated to Petri dishes where the feeding assay is conducted with the larval stages of M. sexta. Damaged leaf tissue is assayed for the expression of relatively early events in signal transduction. Gene expression analysis identified infestation-specific Cys2-His2 (C2H2) transcription factors, confirming the success of using detached leaves in early response studies. The method is easier to perform than whole plant infestations and uses less space.

The presented method creates natural herbivore damaged plant tissue through the application of Manduca sexta larvae to detached leaves of potato. The plant tissue is assayed for expression of six transcription factor homologs involved in early responses to insect herbivory.

Secondo i saggi foglia distaccati per semplificare gli studi sull'espressione genica nelle patate durante l'infestazione da mawing Insect Manduca sexta

The multitrophic nature of gene expression studies of insect herbivory demands large numbers of biological replicates, creating the need for simpler, more ...

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 ...