Ringraziamo Molly saldi per la diluizione e pesticidi assistenza formulazione. Ringraziamo anche Troy Anderson per la guida analisi probit e Zachary Adelman per l&#39;assistenza con disegno sperimentale. Questa ricerca è stata in parte sostenuta da un premio entomologica Foundation e dai fondi borse di studio Virginia Pest Management.

NOTA: Questo protocollo include due saggi insetticidi per il trattamento a parte le uova di cimici e primi stadi. Entrambi i protocolli sono stati condotti utilizzando gli stessi insetticidi, tuttavia, i protocolli dovevano essere adattati per garantire l&#39;esposizione insetticida e di manipolare agevolmente i campioni. 1. Uovo immersione Insetticida Assay <ol><li> Tirare accoppiati, ben nutriti adulti cimici femminili da una colonia e posizionare le femmine (5-10) in una capsula di Petri con carta da filtro. Avere almeno dieci replicati per garantire che ci siano abbastanza uova per il dosaggio insetticidi. Verificare se le femmine hanno deposto le uova ogni giorno. <ol><li> Una volta che depongono le uova, prendere le femmine fuori della scatola di Petri (6 centimetri × 5 cm) e metterli in nuove piastre di Petri con una nuova carta da filtro quotidiano e registrare la data sulle vecchie e nuove piastre di Petri. </li><li> Lasciare le uova a 4 anni a 5 giorni prima di iniziare il test. La suscettibilità agli insetticidi modifiche durante lo sviluppo embrionale, so è importante per il controllo per l&#39;età delle uova. Tenere tutte le fasi della vita a 12:12 L: D fotoperiodo, il 60% di umidità relativa, e 27 ° C. </li></ol></li><li> Una volta che le uova sono invecchiati in modo appropriato, togliere le uova di cimici delicatamente dalla carta da filtro con pinze a punta morbida. raschiare delicatamente la parte inferiore dell&#39;uovo attaccato alla carta da filtro con le pinze e poi afferrare con delicatezza senza applicare molta pressione sull&#39;uovo di limitare lanciando uova di bug letto. </li><li> Mettere le uova di cimici rimossi nei loro rispettivi gruppi all&#39;interno di piatti di plastica Petri. Limitare l&#39;elettricità statica lanciando uova cimici strofinando il fondo di ogni piatto Petri sulla parte esterna con un foglio di ammorbidente. gruppi Trattare di uova di cimici (5-10 uova per gruppo) tutti in una volta. Prendere in considerazione un gruppo di replica e utilizzare almeno cinque (o più) replica per questo test. </li><li> Preparare diluizioni seriali di insetticidi per ottenere una gamma di 5 diverse concentrazioni (0,21-21 microlitri / ml per imidacloprid (0,1%) / β-ciflutrin (0,5%) e un controllosoluzione con il solvente (acqua di rubinetto) solo. Rendere 20 mL di ciascuna concentrazione di avere un volume sufficiente per il dosaggio immersione. </li><li> provette Cut centrifuga (50 mL) alla linea 35 mL medialmente con un utensile rotante con un accessorio disco di taglio e quindi tagliare un foro nel tappo della provetta (diametro = 2.54 cm) usando lo stesso utensile rotante con alluminio ossido di rettifica accessorio di pietra. Lasciare 5 cm sul bordo della calotta durante il taglio, in modo che il tappo si avvitare saldamente nel tubo (Figura 1). </li><li> Tagliare a maglia fine in quadrati grande abbastanza da contenere nel tappo (~ 5 centimetri 2). Avvitare la maglia stretta, eliminando eventuali urti, nelle provette da centrifuga che utilizzano il tappo provetta. </li><li> Posizionare un gruppo di uova di bug letto (una replica) utilizzando un pennello sulla maglia all&#39;interno della provetta. Immergere il gruppo di cimici all&#39;interno della provetta taglio in una concentrazione di insetticida per 5 secondi. </li><li> Rimuovere la provetta da centrifuga di thsoluzione insetticida e. Essiccare la rete con le uova all&#39;interno ponendo la provetta direttamente su un tessuto di laboratorio. </li><li> Rimuovere il gruppo di uova fuori della provetta con un piccolo, fine-setole del pennello e metterli in un ambiente pulito capsula di Petri con una carta da filtro pulito. </li><li> uovo mortalità Record (mancata schiusa) per 14 giorni al fine di garantire tutte le uova hanno avuto il tempo sufficiente per covare. </li><li> Corretta per la mortalità nel trattamento di controllo usando la formula di Abbott (test di mortalità% - controllo% di mortalità / 100 - la mortalità di controllo x 100). </li><li> Inserire la mortalità registrato con le corrispondenti concentrazioni nel software di analisi 15 per calcolare LC 50 valori. </li></ol> 2. Prima Instar Insetticida Assay <ol><li> Separare un gruppo di cimici ben nutriti (5 maschi adulti e 5 femmine adulte), preferibilmente alimentato 1 giorno prima, e metterli in una capsula di Petri (6 centimetri × 5 cm) con carta da filtro previsto mating e la deposizione delle uova. Lasciare le cimici adulti per accoppiarsi e deporre le uova per circa 7-9 giorni. Per l&#39;alimentazione, utilizzare un sistema di alimentazione artificiale che circola acqua calda in tubi di vetro di sangue 16 circostanti. Per questo studio, usare sangue di coniglio defibronated. </li><li> Rimuovere le adulti dalle piastre di Petri (6 cm x 5 cm) e permetta che le uova si schiudono in primi stadi all&#39;interno delle piastre di Petri. Ci vogliono circa 7-9 giorni per le uova di cimici alla schiusa a temperatura ambiente. </li><li> Raccogliere i digiuni primi stadi della stessa età con un pennello. Età può essere determinata rimuovendo i primi stadi quotidianamente e registrando il giorno sono nati. Metterli in nuove, pulite piastre di Petri fino al test insetticidi. Primi ninfe instar non sono molto resistenti essiccante, quindi il saggio insetticida dovrebbe essere completata al più presto possibile. </li><li> Per il saggio, preparare diluizioni seriali di imidacloprid / β-ciflutrin per ottenere una gamma di 5 diverse concentrazioni e una solu controllozione con solo solvente. prodotti formulati etichettati per il controllo di cimici dei letti negli Stati Uniti sono stati utilizzati per questi studi, quindi il solvente era acqua (secondo l&#39;etichetta). </li><li> Pipetta 150 microlitri aliquote di ogni concentrazione insetticida ed acqua per il gruppo di controllo sulle singole carte da filtro (# 1, 4,2 centimetri di diametro). Distribuire l&#39;insetticida sull&#39;intera carta da filtro con il pipettatore applicando goccioline lungo l&#39;intero bordo e il centro della carta da filtro. </li><li> Mettere le carte da filtro trattate individualmente sulla parte superiore dei pannelli di truciolato individuali (7 cm 2). </li><li> gruppi uscita del primo stadi (da 5 a 10 per gruppo) sul singole carte da filtro mentre ancora bagnato con insetticida e coprire i primi stadi con un fondo rovesciata di una capsula di Petri. </li><li> Collocare un peso sopra della scatola di Petri per evitare i primi cimici instar fuoriuscita superficie trattata. </li><li> Record di mortalità dei primi cimici instar dopo 24 ore di esposizione a tuttile concentrazioni di insetticida. </li><li> Corretta per la mortalità nel trattamento di controllo usando la formula di Abbott (test di mortalità% - controllo% di mortalità / 100 - la mortalità di controllo x 100). </li><li> Inserire la mortalità e le concentrazioni registrate nel software di analisi 15 per calcolare LC 50 valori. </li></ol>

<ol>
	<li>Sabou, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bed+bugs+reproductive+life+cycle+in+the+clothes+of+a+patient+suffering+from+Alzheimer's+disease+results+in+iron+deficiency+anemia.">Bed bugs reproductive life cycle in the clothes of a patient suffering from Alzheimer's disease results in iron deficiency anemia.</a> Parasite. 20, (2013).</li><li>Reinhardt, K., Kempke, D., Naylor, R. A., Siva-Jothy, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensitivity+to+bites+by+the+bedbug,+Cimex+lectularius.">Sensitivity to bites by the bedbug, Cimex lectularius.</a> Med. Vet. Entomol. 23, 163-166 (2009).</li><li>Leverkus, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bullous+allergic+hypersensitivity+to+bed+bug+bites+mediated+by+IgE+against+salivary+nitrophorin.">Bullous allergic hypersensitivity to bed bug bites mediated by IgE against salivary nitrophorin.</a> J. invest. entomol. 126, 91-96 (2006).</li><li>Fletcher, C. L., Ardern-Jones, M. R., Hay, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Widespread+bullous+eruption+due+to+multiple+bed+bug+bites.">Widespread bullous eruption due to multiple bed bug bites.</a> Clin. Exp. Dermatol. 27, 74-75 (2002).</li><li>Pritchard, M. J., Hwang, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Severe+anemia+from+bedbugs.">Severe anemia from bedbugs.</a> Can. Med. Assoc. J. 181, 287-288 (2009).</li><li>Goddard, J., de Shazo, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psychological+effects+of+bed+bug+attacks+(Cimex+lectularius+L).">Psychological effects of bed bug attacks (Cimex lectularius L).</a> Am. J. Med. 125, 101-103 (2012).</li><li>Harlan, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bed+bugs+101:+the+basics+of+Cimex+lectularius.">Bed bugs 101: the basics of Cimex lectularius.</a> Am. Entomol. 52, 99-101 (2006).</li><li>Pinto, L. J., Cooper, R., Kraft, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Bed Bug Handbook: The Complete Guide to Bed Bugs and Their Control. , (2007).</li><li>Toloza, A. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+patterns+of+insecticide+resistance+in+eggs+and+first+instars+of+Triatoma+infestans+(Hemiptera:+Reduviidae)+from+Argentina+and+Bolivia.">Differential patterns of insecticide resistance in eggs and first instars of Triatoma infestans (Hemiptera: Reduviidae) from Argentina and Bolivia.</a> J. Med. Entomol. 45, 421-426 (2008).</li><li>Cueto, G. M., Zerba, E. N., Picollo, M. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+of+pyrethroid+resistance+in+eggs+of+Pediculus+humanus+capitis+(Phthiraptera:+Pediculidae)+from+Argentina.">Evidence of pyrethroid resistance in eggs of Pediculus humanus capitis (Phthiraptera: Pediculidae) from Argentina.</a> J. Med. Entomol. 45, 693-697 (2008).</li><li>Ho, S. H., Goh, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deltamethrin+as+a+potential+ovicidal+pyrethroid+against+Plutella+xylostella+L.">Deltamethrin as a potential ovicidal pyrethroid against Plutella xylostella L.</a> Toxicol. let. 22, 161-164 (1984).</li><li>Bell, C. H., Hole, B. D., Evans, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+occurrence+of+resistance+to+phosphine+in+adult+and+egg+stages+of+strains+of+Rhyzopertha+dominica+(F.)(Coleoptera:+Bostrichidae).">The occurrence of resistance to phosphine in adult and egg stages of strains of Rhyzopertha dominica (F.)(Coleoptera: Bostrichidae).</a> J. Stored Prod. Res. 13, 91-94 (1977).</li><li>Campbell, B. E., Miller, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insecticide+Resistance+in+Eggs+and+First+Instars+of+the+Bed+Bug,+Cimex+lectularius+(Hemiptera:+Cimicidae).">Insecticide Resistance in Eggs and First Instars of the Bed Bug, Cimex lectularius (Hemiptera: Cimicidae).</a> Insects. 6, 122-132 (2015).</li><li>Price, L. A., Mills, K. 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+toxicity+of+phosphine+to+the+immature+stages+of+resistant+and+susceptible+strains+of+some+common+stored+product+beetles,+and+implications+for+their+control.">The toxicity of phosphine to the immature stages of resistant and susceptible strains of some common stored product beetles, and implications for their control.</a> J. Stored Prod. Res. 24, 51-59 (1988).</li><li>Robertson, J. L., Preisler, H. K., Russel, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> PoloPlus: Probit and Logit Analysis User's Guide. , (2003).</li><li>Montes, C., Cuadrillero, C., Vilella, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+of+a+laboratory+colony+of+Cimex+lectularius+(Hemiptera:+Cimicidae)+using+an+artificial+feeding+technique.">Maintenance of a laboratory colony of Cimex lectularius (Hemiptera: Cimicidae) using an artificial feeding technique.</a> J. Med. Entomol. 39, 675-679 (2002).</li><li>Brogdon, W. G., McAllister, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplification+of+adult+mosquito+bioassays+through+use+of+time-mortality+determinations+in+glass+bottles.">Simplification of adult mosquito bioassays through use of time-mortality determinations in glass bottles.</a> J. Am. Mosq. Control. Assoc. 14, 159-164 (1998).</li><li>Katarzyna, K., Saddler, A., Koella, J. C. <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+age+and+larval+nutrition+on+phenotypic+expression+of+insecticide-resistance+in+Anopheles+mosquitoes.">Effects of age and larval nutrition on phenotypic expression of insecticide-resistance in Anopheles mosquitoes.</a> PLoS One. 8 (3), (2013).</li><li>Outram, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+affecting+the+resistance+of+insect+eggs+to+sulphuryl+fluoride-II:+the+distribution+of+sulphuryl-35+S+fluoride+in+insect+eggs+after+fumigation.">Factors affecting the resistance of insect eggs to sulphuryl fluoride-II: the distribution of sulphuryl-35 S fluoride in insect eggs after fumigation.</a> J. Stored. Prod. Res. 3, 353-358 (1967).</li></ol>

Gli autori non hanno nulla da rivelare.

A critical step in this assay is to ensure that no eggs that are removed from a surface are damaged prior to the assay. Many insects cement their eggs to a substrate, therefore, a preliminary test may be needed to ensure that removal does not cause mortality. This test can be conducted with several replications of a treated group (bed bugs removed from filter paper) compared to a control group (bed bugs not removed). Similar low rates of mortality (or no mortality) between the treated and control group will ensure that ...

Le cimici dei letti sono un parassita urbano significativo che può causare la perdita di sangue, irritazioni della pelle, insonnia, depressione e ansia nei loro ospiti umani 1, 2, 3, 4, 5, 6. I costi di smaltimento e controllare le cimici dei letti sono alti e spesso richiedono più visite a casa da una società di controllo dei parassiti 7. visite multiple sono di solito necessari a causa del comportamento criptico di cimici e le difficoltà connesse con la loro uccisione. In particolare, le uova di cimici sono difficili da controllare a causa della loro piccola dimensione e la protezione dell&#39;embrione dal guscio. Attualmente, una pratica comune per molte aziende di controllo dei parassiti in tutti gli Stati Uniti è quello di trattare una casa per le cimici dei letti utilizzando un insetticida chimico. È risaputoche le uova sono difficili da controllare, così molte aziende hanno implementato un lasso di tempo di due settimane per ri-trattamenti 8. Questo permette di uova di cimici abbastanza tempo per covare, in modo che il primo stadio emergerà e presumibilmente più facile per uccidere con insetticidi. Tuttavia, vi è una carenza di studi che valutano l&#39;efficacia degli insetticidi liquido contro le uova di bug letto e primi stadi. uova di insetti sono stati documentati per essere il più difficile fase della vita da controllare in insetti infestanti urbani, diverso da cimici dei letti, oltre a molti parassiti agricoli. La maggior parte di queste difficoltà sono state attribuite al guscio d&#39;uovo, mentre pochi studi hanno riportato resistenza agli insetticidi. Resistenza allo stadio di uovo è stato documentato in Triatoma infestans 9, Pediculus humanus capitis 10, Plutella xylostella 11, Rhyzopertha dominica 12, <em&gt; Cimex lectularius 13, e più coleotteri di prodotti conservati (cioè Oryzaephilus surinamensis, Tribolium castaneum, Cryptolestes ferrugineus e Rhyzopertha dominica). 14 Lo sviluppo della resistenza nella fase di uovo e stadi immaturi, così come le complicazioni associate con la penetrazione insetticida attraverso il guscio d&#39;uovo, richiede la necessità di test biologici insetticida efficacia contro queste fasi della vita. Questo protocollo prevede una procedura passo-passo per determinare l&#39;efficacia di insetticidi nell&#39;uovo e primo stadio fase di cimici comuni, Cimex lectularius. Entrambi questi protocolli sono saggi concentrazione-risposta per consentire la quantificazione di LC 50 valori. saggi di concentrazione-risposta sono comunemente usati per studi tossicologici, ma questo protocollo è stato adattato per il trattamento facilmente gruppi di uova di bug letto e primi stadi. Questi saggi possono essere adattatiper le uova diverse specie di insetti &#39;e le fasi della vita immaturi.

<table border="0" cellpadding="0" cellspacing="0" width="1067">
	<tbody>
		<tr>
			<td>Petri dishes</td>
			<td>Fisher Scientific Inc.</td>
			<td>08-757-105</td>
			<td>Plastic</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>Whatman</td>
			<td>1001-042</td>
			<td></td>
		</tr>
		<tr>
			<td>Featherweight forceps</td>
			<td>Fisher Scientific Inc.</td>
			<td align="right">4750</td>
			<td></td>
		</tr>
		<tr>
			<td>Temprid SC (imidacloprid/bifenthrin)</td>
			<td>Bayer CropScience</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Transport GHP (acetamiprid/bifenthrin)</td>
			<td>FMC Corp.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge tube</td>
			<td></td>
			<td>06-443-18</td>
			<td>50 mL; flat-top threaded</td>
		</tr>
		<tr>
			<td>Fine mesh</td>
			<td></td>
			<td>7250C</td>
			<td></td>
		</tr>
		<tr>
			<td>KimWipe</td>
			<td></td>
			<td>06-666A</td>
			<td>11 cm X 21 cm</td>
		</tr>
		<tr>
			<td>Small paint brush</td>
			<td></td>
			<td></td>
			<td>Any small paint brush</td>
		</tr>
		<tr>
			<td>Hardboard panels</td>
			<td></td>
			<td></td>
			<td>Composite wood or tile would be sufficient from a home improvement store</td>
		</tr>
		<tr>
			<td>Metal weights</td>
			<td></td>
			<td></td>
			<td>Any type of metal screw that will hold the weight of the Petri dish down onto a surface</td>
		</tr>
	</tbody>
</table>

a method for evaluating insecticide efficacy against bed bug, cimex lectularius, eggs and first instars

valutazioni di tossicità standard di insetticidi contro insetti nocivi sono condotte principalmente di insetti adulti. Le valutazioni sono basate su una dose-risposta o curva concentrazione-risposta, se la mortalità aumenta all&#39;aumentare della dose o concentrazione di un insetticida è aumentato. Concentrazione standard letale (LC 50) e dose letale (LD 50) prove che si traducono nel 50% di mortalità di una popolazione di prova può essere difficile per valutare la tossicità degli insetticidi contro i non-adulti fasi di vita degli insetti, come le uova e instar presto o fasi nymphal . Tuttavia, queste informazioni sono essenziali per la comprensione di efficacia insetticida in tutte le fasi della vita di cimici, che colpisce gli sforzi di controllo e di trattamento. Questo protocollo utilizza un saggio biologico immersione standard modificato per le uova di cimici e un test di insetticida di contatto per il trattamento di nymphal primi stadi. Questi test producono una curva di concentrazione-risposta per quantificare ulteriormente LC 50 valori per le valutazioni insetticida.

Le uova sono stati immersi in 5 diverse concentrazioni di imidacloprid / β-ciflutrin (ul / mL). I primi stadi sono stati collocati su superfici trattate di cinque diverse concentrazioni di imidacloprid / β-ciflutrin (Figura 2). Abbiamo usato tre diverse popolazioni di C. lectularius: Harlan, Richmond, ed Epic Centro (Tabella 1). Il ceppo Harlan è suscettibile di insett...

Watch this Scientific Journal Video about A Method for Evaluating Insecticide Efficacy against Bed Bug,  Cimex lectularius, Eggs and First Instars at JoVE.com

A Method for Evaluating Insecticide Efficacy against Bed Bug,  Cimex lectularius, Eggs and First Instars

valutazioni Insetticida sono spesso presi di mira contro gli insetti adulti, piuttosto che stadi immaturi. Qui, vi presentiamo un protocollo per la valutazione insetticidi contro le uova di bug letto con un confronto con il primo stadio bug letto ninfale. Questi protocolli possono essere regolati per altri insetti per valutare l&#39;efficacia insetticida in fasi della vita non adulti.

Un metodo per la valutazione Insetticida efficacia contro Bed Bug,<em&gt; Cimex lectularius</em&gt;, Uova e First stadi

Standard toxicity evaluations of insecticides against insect pests are primarily conducted on adult insects. Evaluations are based on a dose-response or ...

a-method-for-evaluating-insecticide-efficacy-against-bed-bug-cimex

Research

JoVE Journal

Environment

A Method for Evaluating Insecticide Efficacy against Bed Bug, Cimex lectularius, Eggs and First Instars

11.2K Views.  University of Florida.  valutazioni Insetticida sono spesso presi di mira contro gli insetti adulti, piuttosto che stadi immaturi. Qui, vi presentiamo un protocollo per la valutazione insetticidi contro le uova di bug letto con un confronto con il primo stadio bug letto ninfale. Questi protocolli possono essere regolati per altri insetti per valutare l'efficacia insetticida in fasi della vita non adulti. 

Video: A Method for Evaluating Insecticide Efficacy against Bed Bug,  Cimex lectularius, Eggs and First Instars

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is a biotrophic fungal pathogen and the causal agent of corn smut on maize. This disease is responsible for significant yield losses of approximately $1.0 billion annually in the U.S.1 Several methods including crop rotation, fungicide application and seed treatments are currently used to control corn smut2. However, host resistance is the only practical method for managing corn smut. Identification of crop plants including maize, wheat, and rice that are resistant to various biotrophic pathogens has significantly decreased yield losses annually3-5. Therefore, the use of a pathogen inoculation method that efficiently and reproducibly delivers the pathogen in between the plant leaves, would facilitate the rapid identification of maize lines that are resistant to U. maydis. As, a first step toward indentifying maize lines that are resistant to U. maydis, a needle injection inoculation method and a resistance reaction screening method was utilized to inoculate maize, teosinte, and maize x teosinte introgression lines with a U. maydis strain and to select resistant plants.
Maize, teosinte and maize x teosinte introgression lines, consisting of about 700 plants, were planted, inoculated with a strain of U. maydis, and screened for resistance. The inoculation and screening methods successfully identified three teosinte lines resistant to U. maydis. Here a detailed needle injection inoculation and resistance reaction screening protocol for maize, teosinte, and maize x teosinte introgression lines is presented. This study demonstrates that needle injection inoculation is an invaluable tool in agriculture that can efficiently deliver U. maydis in between the plant leaves and has provided plant lines that are resistant to U. maydis that can now be combined and tested in breeding programs for improved disease resistance.

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

The use of a needle injection method to inoculate maize and teosinte plants with the biotrophic pathogen Ustilago maydis is described. The needle injection inoculation method facilitates the controlled delivery of the fungal pathogen in between the plant leaves where the pathogen enters the plant through the formation of appresoria. This method is highly efficient, enabling reproducible inoculations with U. maydis.

Un metodo rapido ed efficiente per valutare la patogenicità dei maydis di Ustilago sulle linee di mais e teosinte

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is ...

Ricerca

Ambiente

A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and enables to perform rapid, automated and simple SPE. The pre-concentrated solution is compatible with analysis by immunoassay, with a low organic solvent content. A method is described for the extraction and pre-concentration of natural hormone 17&#946;-estradiol in 100 ml water samples. Reverse phase SPE is performed with octadecyl-silica sorbent and elution is done with 200 &#181;l of methanol 50% v/v. Eluent is diluted by adding di-water to lower the amount of methanol. After preparing manually the SPE column, the overall procedure is performed automatically within 1 hr. At the end of the process, estradiol concentration is measured by using a commercial enzyme-linked immune-sorbent assay (ELISA). 100-fold pre-concentration is achieved and the methanol content in only 10% v/v. Full recoveries of the molecule are achieved with 1 ng/L spiked de-ionized and synthetic sea water samples.

A protocol for the extraction and pre-concentration of estradiol from water samples by using an automated and miniaturized system is presented.

Un metodo semplice per Automated fase solida estrazione di campioni di acqua per l&#39;analisi immunologiche di piccole inquinanti

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and ...

Use of a Battery of Chemical and Ecotoxicological Methods for the Assessment of the Efficacy of Wastewater Treatment Processes to Remove Estrogenic Potency

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in a watch list of environmental pollutants under the European Water Framework Directive. Municipal wastewater treatment plants are major contributors to the estrogenic potency of surface waters. Much of the estrogenic potency of wastewater treatment plant (WWTP) effluents can be attributed to the discharge of steroid estrogens including estradiol (E2), EE2 and E1 due to incomplete removal of these substances at the treatment plant. An evaluation of the efficacy of wastewater treatment processes requires the quantitative determination of individual substances most often undertaken using chemical analysis methods. Most frequently used methods include Gas Chromatography-Mass Spectrometry (GCMS/MS) or Liquid Chromatography-Mass Spectrometry (LCMS/MS) using multiple reaction monitoring (MRM). Although very useful for regulatory purposes, targeted chemical analysis can only provide data on the compounds (and specific metabolites) monitored. Ecotoxicology methods additionally ensure that any by-products produced or unknown estrogenic compounds present are also assessed via measurement of their biological activity. A number of in vitro bioassays including the Yeast Estrogen Screen (YES) are available to measure the estrogenic activity of wastewater samples. Chemical analysis in conjunction with in vivo and in vitro bioassays provides a useful toolbox for assessment of the efficacy and suitability of wastewater treatment processes with respect to estrogenic endocrine disrupting compounds. This paper utilizes a battery of chemical and ecotoxicology tests to assess conventional, advanced and emerging wastewater treatment processes in laboratory and field studies.

Endocrine Disrupting Compounds (EDC) pose a substantial risk to the aquatic environment. Municipal wastewater treatment plants are major contributors to the estrogenic potency of surface waters. The methodology provided in this paper allows for an assessment of the efficacy and suitability of wastewater treatment processes with respect to EDC removal.

Utilizzo di una batteria di chimici ed ecotossicologici metodi per la valutazione dell&#39;efficacia dei processi di trattamento delle acque reflue per rimuovere estrogenica Potenza

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in ...

A Gusseted Thermogradient Table to Control Soil Temperatures for Evaluating Plant Growth and Monitoring Soil Processes

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a series of incubators. A temperature gradient is passively established across the surface of the table between the heated and cooled ends and is lost quickly at distances above the surface. Since temperature is only controlled on the table surface, experiments are restricted to shallow containers, such as Petri dishes, placed on the table. Welding continuous aluminum vertical strips or gussets perpendicular to the surface of a table enables temperature control in depth via convective heat flow. Soil in the channels between gussets was maintained across a gradient of temperatures allowing a greater diversity of experimentation. The gusseted design was evaluated by germinating oat, lettuce, tomato, and melon seeds. Soil temperatures were monitored using individual, battery-powered dataloggers positioned across the table. LED lights installed in the lids or along the sides of the gradient table create a controlled temperature chamber where seedlings can be grown over a range of temperatures. The gusseted design enabled accurate determination of optimum temperatures for fastest germination rate and the highest percentage germination for each species. Germination information from gradient table experiments can help predict seed germination and seedling growth under the adverse soil conditions often encountered during field crop production. Temperature effects on seed germination, seedling growth, and soil ecology can be tested under controlled conditions in a laboratory using a gusseted thermogradient table.

Traditional thermogradient tables create a range of temperatures across the surface. Welding gussets perpendicular to the surface of a thermogradient table will control temperature in depth increasing possible research applications.

Una tabella soffietto Thermogradient di controllo della temperatura del suolo per la valutazione di crescita delle piante e monitoraggio processi del suolo

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a ...

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester fabric and glue can be folded and glued to construct a strip of compartments that house seeds and identifying information, while allowing contact with soil leachate, water, microorganisms, and ambient temperature. Strips may be constructed with a wide range of compartment numbers and sizes and allow the researcher to house a variety of genotypes within a single species, different species, or seeds that have experienced different treatments. As opposed to individual seed packets, strips are more easily retrieved as a unit. While replicate packets can be included within a strip, different strips can act as blocks or can be retrieved at different times for observation of seed behavior over time. We used a high temperature glue gun to delineate compartments and sealed the strips once the seed and tags identifying block and removal times were inserted. The seed strips were then buried in the field at the desired depth, with the location marked for later removal. Burrowing animal predators were effectively excluded by use of a covering of metal mesh hardware cloth on the soil surface. After the selected time interval for burial, strips were dug up and seeds were assessed for germination, dormancy and mortality. While clearly dead seeds can often be distinguished from ungerminated living ones by eye, dormant seeds were conclusively identified using a standard Tetrazolium chloride colorimetric test for seed viability.

Here we present a protocol for assessing seed survivorship, germination and dormancy under field conditions using buried, labeled seed strips and tetrazolium chloride (TZ) viability testing.

Metodo affidabile per valutare la germinazione dei semi, dormienza, e mortalità in normali condizioni operative

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

Un sistema di esposizione di flusso-through per valutare sospeso Sedimenti Effetti sulla vita acquatica

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples

Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial communities, a relatively precise but effort-intensive technique to assay microbial community composition is phospholipid fatty acid (PLFA) analysis. PLFA was developed to analyze phospholipid biomarkers, which can be used as indicators of microbial biomass and the composition of broad functional groups of fungi and bacteria. It has commonly been used to compare soils under alternative plant communities, ecology, and management regimes. The PLFA method has been shown to be sensitive to detecting shifts in microbial community composition.
An alternative method, fatty acid methyl ester extraction and analysis (MIDI-FA) was developed for rapid extraction of total lipids, without separation of the phospholipid fraction, from pure cultures as a microbial identification technique. This method is rapid but is less suited for soil samples because it lacks an initial step separating soil particles and begins instead with a saponification reaction that likely produces artifacts from the background organic matter in the soil. 
This article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples. The method combines the accuracy achieved through PLFA profiling by extracting and concentrating soil lipids as a first step, and a reduction in effort by saponifying the organic material extracted and processing with the MIDI-FA method as a second step.

The article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples.

Un metodo di estrazione e analisi lipidica per la caratterizzazione dei microbi del suolo negli esperimenti con molti campioni

Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial ...

Image-based Lagrangian Particle Tracking in Bed-load Experiments

Image analysis has been increasingly used for the measurement of river flows due to its capabilities to furnish detailed quantitative depictions at a relatively low cost. This manuscript describes an application of particle tracking velocimetry (PTV) to a bed-load experiment with lightweight sediment. The key characteristics of the investigated sediment transport conditions were the presence of a covered flow and of a fixed rough bed above which particles were released in limited number at the flume inlet. Under the applied flow conditions, the motion of the individual bed-load particles was intermittent, with alternating movement and stillness terms. The flow pattern was preliminarily characterized by acoustic measurements of vertical profiles of the stream-wise velocity. During process visualization, a large field of view was obtained using two action-cameras placed at different locations along the flume. The experimental protocol is described in terms of channel calibration, experiment realization, image pre-processing, automatic particle tracking, and post-processing of particle track data from the two cameras. The presented proof-of-concept results include probability distributions of the particle hop length and duration. The achievements of this work are compared to those of existing literature to demonstrate the validity of the protocol.

The manuscript presents a protocol for the conduction of bed-load sediment transport experiments where the moving particles are tracked by image analysis. The experimental facility, the procedures for run realization and data processing, and finally some proof-of-concept results are presented here.

Tracciamento delle particelle lagrangiche basate sull&#39;immagine in esperimenti di Bed-load

Image analysis has been increasingly used for the measurement of river flows due to its capabilities to furnish detailed quantitative depictions at a ...

A Novel Method for the Pentosan Analysis Present in Jute Biomass and Its Conversion into Sugar Monomers Using Acidic Ionic Liquid

Recently, ionic liquids (ILs) are used for biomass valorization into valuable chemicals because of their remarkable properties such as thermal stability, lower vapor pressure, non-flammability, higher heat capacity, and tunable solubility and acidity. Here, we demonstrate a method for the synthesis of C5 sugars (xylose and arabinose) from the pentosan present in jute biomass in a one-pot process by utilizing a catalytic amount of Br&#248;nsted acidic 1-methyl-3-(3-sulfopropyl)-imidazolium hydrogen sulfate IL. The acidic IL is synthesized in the lab and characterized using NMR spectroscopic techniques for understanding its purity. The various properties of BAIL are measured such as acid strength, thermal and hydrothermal stability, which showed that the catalyst is stable at a higher temperature (250 &#176;C) and possesses very high acid strength (Ho 1.57). The acidic IL converts over 90% of pentosan into sugars and furfural. Hence, the presenting method in this study can also be employed for the evaluation of pentosan concentration in other kinds of lignocellulosic biomass.

We present a protocol for the synthesis of C5 sugars (xylose and arabinose) from a renewable non-edible lignocellulosic biomass (i.e., jute) with the presence of Br&#248;nsted acidic ionic liquids (BAILs) as the catalyst in water. The BAILs catalyst exhibited better catalytic performance than conventional mineral acid catalysts (H2SO4 and HCl).

Un nuovo metodo per l'analisi di pentosano presente nella biomassa di iuta e la relativa conversione in monomeri di zucchero utilizzando acido liquido ionico

Recently, ionic liquids (ILs) are used for biomass valorization into valuable chemicals because of their remarkable properties such as thermal stability, ...

Visualizing Efficacy of Pesticides Against Disease Vector Mosquitoes in the Field

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform across ecological zones. To best protect public and veterinary health from these insects, the environmental limitations of pesticides need to be investigated to inform effective use of the most appropriate pesticide formulations and techniques. We have developed a research program to evaluate combinations of pesticides, pesticide application equipment, and application techniques in hot-arid desert, hot-humid tropical, warm and cool temperate, and urban locations to derive pesticide use guidelines specific to target insect and environment. To these ends we designed a system of protocols to support efficient, cost-effective, portable, and standardized evaluation of a diverse range of pesticides and equipment across multiple environments. At the core of these protocols is the use of an array of small cages with colony-reared sentinel mosquitoes (adults and immatures) and sand flies (adults), strategically arranged in natural habitats and exposed to pesticide spray. Spatial and temporal patterns of pesticide efficacy are derived from percent mortality in sentinel cages, then mapped and visualized in a geographic information system. Maps of sentinel mortality data may be statistically compared to evaluate relative efficacy of a pesticide across multiple environments, or to study multiple pesticides in a single environment. Protocols may be modified to accommodate a variety of scenarios, including, for example, the vertical orientation of sentinels in canopy habitats or simultaneous testing of ground and aerial application methods.

The efficacy of public health pesticides targeting nuisance and disease-vector insects is not uniform across different ecological zones. Here we present a system of techniques using captive vector insects as sentinels for pesticide efficacy to derive electronic maps supporting the standard evaluation of pesticides across multiple environments.

Efficacia dei pesticidi contro le zanzare vettore di malattia nel campo di visualizzazione

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform ...