Wir danken Molly Stedfast zur Verdünnung und Pestizid-Formulierung Unterstützung. Wir danken auch Troy Anderson für Probit-Analyse Führung und Zachary Adelman für die Unterstützung bei experimentellen Design. Diese Forschung wurde teilweise durch eine Entomologischen Foundation Award und von Virginia Pest Management Stipendienfonds unterstützt.

HINWEIS: Dieses Protokoll enthält zwei Insektizid-Assays für separat Bettwanze Eier und erste instars behandeln. Beide Protokolle wurden die gleichen Insektiziden durchgeführt unter Verwendung von, jedoch mussten die Protokolle angepasst werden Insektizid Exposition und zu leicht, die Proben zu manipulieren. 1. Egg Dipping Insecticide Assay <ol><li> Ziehen gepaart, gut gefütterten erwachsenen weiblichen Bettwanzen aus einer Kolonie, und legen die Weibchen (5-10) in eine Petrischale mit Filterpapier. Mindestens zehn Replikaten, um sicherzustellen, dass es genug Eier für das Insektizid-Test sein. Überprüfen Sie, ob die Weibchen jeden Tag Eier gelegt haben. <ol><li> Sobald sie Eier legen, nehmen die Weibchen aus der Petrischale (6 cm x 5 cm) und sie in neue Petrischalen mit einem neuen Filterpapier täglich und das Datum, an dem alten und neuen Petrischalen aufnehmen. </li><li> Lassen Sie die Eier im Alter von 4 bis 5 Tage vor Beginn des Tests. Die Empfindlichkeit gegenüber Insektiziden Veränderungen während der embryonalen Entwicklung, so ist es wichtig, für die Eier Alter zu steuern. Halten Sie alle Lebensphasen um 12:12 L: D-Photoperiode, 60% relativer Luftfeuchtigkeit und 27 ° C. </li></ol></li><li> Sobald die Eier entsprechend gealtert sind, entfernen Sie die Bettwanze Eier vorsichtig von Filterpapiere Soft-Spitze Pinzette. Vorsichtig kratzen den Boden des Ei auf das Filterpapier angebracht mit der Pinzette und dann ohne Anwendung viel Druck auf das Ei zu werfen Bettwanzen Eier zu begrenzen sanft greifen. </li><li> Die ausgebaute Bettwanzen Eier in ihren jeweiligen Gruppen innerhalb von Petrischalen aus Kunststoff. Begrenzen Sie die statische Elektrizität von Bettwanzen Eier schleudert durch den Boden jeder Petrischale reiben auf der Außenseite mit einem Weichspüler Blatt. Treat Gruppen von Bettwanzen Eier (5-10 Eier pro Gruppe) alle auf einmal. Betrachten wir eine Gruppe, die eine Replikation und verwenden mindestens fünf (oder mehr) repliziert für diesen Test. </li><li> Bereiten Reihenverdünnungen von Insektiziden eine Reihe von 5 verschiedenen Konzentrationen zu erhalten (0,21 bis 21 &amp; mgr; l / ml für Imidacloprid (0,1%) / β-Cyfluthrin (0,5%) und eine SteuerLösung mit Lösungsmittel (Leitungswasser) nur. Machen Sie 20 ml jeder Konzentration genügend Volumen für den Tauch Test zu haben. </li><li> Cut-Zentrifugenröhrchen (50 ml) in der 35 ml Linie median mit einem Drehwerkzeug mit einer Trennscheibe Zubehör und dann ein Loch in den Deckel des Zentrifugenrohr (Durchmesser = 2,54 cm) mit dem gleichen Drehwerkzeug mit einem Aluminium geschnitten Oxid Schleifstein Zubehör. Lassen Sie 5 cm am Rand der Kappe beim Schneiden, so dass die Kappe fest in dem Rohr (Abbildung 1) Schraube wird. </li><li> Schneiden Sie feinmaschigen in Quadrate groß genug , um in die Kappe zu passen (~ 5 cm 2). Schrauben Sie das Gitter fest, alle Unebenheiten zu beseitigen, in die Zentrifugenröhrchen Zentrifugenröhrchen Kappe mit. </li><li> Legen Sie eine Gruppe von Bettwanzen Eier (eine Wiederholung) einen Pinsel auf das Netz innerhalb des Zentrifugenröhrchen verwenden. Tauchen Sie die Gruppe von Bettwanzen im Inneren des geschnittenen Zentrifugenröhrchen in eine Insektizidkonzentration für 5 Sekunden. </li><li> Entfernen Sie die Zentrifugenröhrchen aus the Insektizidlösung. Trocknen Sie das Gitter mit den Eiern im Inneren durch das Zentrifugenröhrchen direkt auf ein Labor Gewebe platzieren. </li><li> Entfernen Sie die Gruppe von Eiern aus dem Zentrifugenröhrchen einen kleinen, fein Borste Pinsel und legen Sie sie in eine saubere Petrischale mit einem sauberen Filterpapier. </li><li> Nehmen Sie Ei-Sterblichkeit (Ausfall zu schlüpfen) für 14 Tage alle Eier, um sicherzustellen, haben genügend Zeit zu schlüpfen musste. </li><li> Richtige für die Mortalität in der Kontrollbehandlung Abbott-Formel (% Test Mortalität -% Kontrollmortalität / 100 - Kontrollmortalität x 100). </li><li> Legen Sie aufgezeichnet Mortalität im Zusammenhang mit den entsprechenden Konzentrationen in die Analysesoftware 15 LC 50 Werte zu berechnen. </li></ol> 2. Erste Instar Insecticide Assay <ol><li> Trennen Sie eine Gruppe von gut genährten Bettwanzen (5 erwachsene Männer und 5 erwachsene Frauen), vorzugsweise 1 Tag vor gefüttert, und legen Sie sie in eine Petrischale (6 cm x 5 cm) mit Filterpapier für matin vorgeseheng und Eiablage. Lassen Sie die erwachsenen Bettwanzen Eier zu paaren und legen für etwa 7-9 Tage. Zur Fütterung verwenden , um eine künstliche Ernährung System , das Warmwasser in Glasröhrchen umgebenden Blut 16 zirkuliert. Für diese Studie verwenden defibronated Kaninchenblut. </li><li> Entfernen Sie die Erwachsenen von den Petrischalen (6 cm x 5 cm) und lassen Sie die Eier in die erste instars in den Petrischalen zu schlüpfen. Es dauert ungefähr 7-9 Tage für Eier Bettwanze bei Raumtemperatur zu schlüpfen. </li><li> Sammeln Sie die unfed ersten instars des gleichen Alters mit einem Pinsel. Das Alter kann durch Entfernen der ersten instars täglich bestimmt werden und die Aufzeichnung der Tag sie ausgebrütet. Legen Sie sie in neue, saubere Petrischalen bis zum Insektizid-Test. Erste instar Nymphen sind nicht sehr Trocknungs resistent, so sollte das Insektizid-Test so schnell wie möglich abgeschlossen werden. </li><li> Für den Test vorbereiten Reihenverdünnungen von Imidacloprid / β-Cyfluthrin eine Reihe von 5 verschiedenen Konzentrationen und eine Steuer solu zu bekommention nur mit Lösungsmittel. Formulierte Produkte für Bettwanze Kontrolle in den Vereinigten Staaten markiert wurden für diese Untersuchungen verwendet daher das Lösungsmittel Wasser (nach dem Etikett). </li><li> Pipette 150 ul Aliquots jeder Insektizidkonzentration und Wasser für die Kontrollgruppe auf einzelnen Filterpapiere (# 1; 4,2 cm Durchmesser). das Insektizid über das gesamte Filterpapier verteilen sich um den gesamten Rand und in der Mitte des Filterpapiers mit der Pipettiervorrichtung Tröpfchen durch Anwendung. </li><li> Legen Sie die behandelten Filterpapiere individuell auf einzelne Hartfaserplatten (7 cm 2). </li><li> Veröffentlichung Gruppen erster instars (5 bis 10 pro Gruppe) auf die einzelnen Filterpapiere, während noch nass mit Insektizid und decken die erste instars mit einem umgekehrten Boden einer Petrischale. </li><li> Legen Sie ein Gewicht auf der Petrischale, die ersten instar Bettwanzen zu verhindern, dass die behandelte Oberfläche zu entkommen. </li><li> Die Mortalität der ersten instar Bettwanzen nach 24 h Exposition gegenüber allenInsektizid-Konzentrationen. </li><li> Richtige für die Mortalität in der Kontrollbehandlung Abbott-Formel (% Test Mortalität -% Kontrollmortalität / 100 - Kontrollmortalität x 100). </li><li> Legen Sie aufgezeichnet Mortalität und Konzentrationen in die Analysesoftware 15 LC 50 Werte zu berechnen. </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>

Die Autoren haben nichts zu offenbaren.

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 ...

Bettwanzen sind eine bedeutende städtische Schädlingsbekämpfung , die Blutverlust verursachen können, Hautreizungen, Schlaflosigkeit, Depression und Angst in ihren menschlichen Wirten 1, 2, 3, 4, 5, 6. Die Kosten für die Beseitigung und Kontrolle Bettwanzen sind hoch und oft erfordern mehrere Besuche in einem Haus von einem Schädlingsbekämpfung Unternehmen 7. Mehrere Besuche sind in der Regel wegen des kryptischen Verhalten von Bettwanzen erforderlich und die Schwierigkeiten, die mit ihnen zu töten. Insbesondere sind Bettwanzen Eier schwierig wegen ihrer geringen Größe zu kontrollieren und den Schutz des Embryos durch die Eischale. Derzeit ist eine gängige Praxis für viele Schädlingsbekämpfung Unternehmen in den Vereinigten Staaten ein Haus für Bettwanzen zu behandeln, ein chemisches Insektizid verwendet wird. Es ist allgemein Bekanntdass die Eier schwer zu kontrollieren sind, haben so viele Unternehmen 8 einen zweiwöchigen Zeitrahmen für die Wieder Behandlungen umgesetzt. Auf diese Weise können Bettwanzen Eier genug Zeit zu schlüpfen, so dass das erste Larvenstadium entstehen und angeblich leichter sein, mit Insektiziden zu töten. Allerdings gibt es einen Mangel an Studien, die die Wirksamkeit von flüssigen Insektiziden gegen Bettwanzen Eier und erste instars Auswertung. Insekteneier wurden die schwierigsten Lebensphase zu steuern, in städtischen Schadinsekten, andere als Bettwanzen, zusätzlich zu den vielen landwirtschaftlichen Schädlingen zu dokumentieren. Die meisten dieser Schwierigkeiten wurden in der Eierschale zurückgeführt, während einige Studien Insektizidresistenz berichteten. Der Widerstand im Ei - Stadium hat in Triatoma infestans 9, Pediculus humanus capitis 10 dokumentiert, Plutella xylostella 11, Rhyzopertha dominica 12, <em&gt; Cimex lectularius 13 und mehrere gespeicherte Produkt Käfer (dh Getreideplattkäfer, Tribolium castaneum, Cryptolestes ferrugineus und Rhyzopertha dominica). 14 Die Entwicklung des Widerstandes im Ei - Stadium und unreifen Stadien sowie mit Insektizid Penetration durch die Eischale verbundenen Komplikationen, erfordert die Notwendigkeit Insektizid Wirksamkeit Biotests gegen diese Lebensphasen. Dieses Protokoll ein Verfahren Schritt- für -Schritt stellt die Wirksamkeit von Insektiziden im Ei und erste Larvenstadium Stadium der gemeinsamen Bettwanzen zur Bestimmung, Cimex lectularius. Beide Protokolle sind Konzentrations-Wirkungs - Assays Quantifizierung der LC 50 -Werte zu ermöglichen. Konzentrations-Wirkungs-Assays sind für toxikologische Studien häufig verwendet, aber dieses Protokoll für die einfache Behandlung von Gruppen von Bettwanzen Eier und erste instars angepasst wurde. Diese Assays können angepasst werden,für verschiedene Insektenarten Eier und unreifen Lebensphasen.

<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

Standard-Toxizität Auswertungen von Insektiziden gegen Insektenschädlinge sind auf erwachsene Insekten in erster Linie durchgeführt. Auswertungen basieren auf einer Dosis-Wirkungs-oder Konzentrations-Wirkungs-Kurve, wo die Mortalität erhöht, wenn die Dosis oder Konzentration eines Insektizids erhöht wird. Standard - letale Konzentration (LC 50) und letale Dosis (LD 50) Tests , die in 50% Sterblichkeit einer Testpopulation führen kann zur Auswertung Toxizität von Insektiziden gegen nicht ausgewachsene Tier Lebensphasen eine Herausforderung sein, wie Eier und frühen Larvenstadium oder Nymphenstadien . Allerdings ist diese Information für das Verständnis Insektizid Wirksamkeit in allen Bettwanze Lebensphasen, die die Bemühungen um Kontrolle und Behandlung betrifft. Dieses Protokoll verwendet ein Standard-Bioassay Tauchen für Bettwanzen Eier modifiziert und einen Kontakt insektizide Assay für nymphal erste instars Behandlung. Diese Assays erzeugen eine Konzentrations-Wirkungs - Kurve zu weiteren LC 50 -Werte für Insektizid Auswertungen zu quantifizieren.

Die Eier wurden eingetaucht in 5 verschiedenen Konzentrationen von Imidacloprid / β-Cyfluthrin (ul / ml). Die ersten instars wurden auf behandelten Oberflächen von fünf verschiedenen Konzentrationen von Imidacloprid / β-Cyfluthrin (Figur 2) plaziert. Wir haben drei verschiedene Populationen von C. lectularius: Harlan, Richmond, und epische Center (Tabelle 1). Die Harl...

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

Insecticide Evaluierungen werden häufig gegen erwachsenen Insekten gezielt, anstatt unreifen Stadien. Hier präsentieren wir ein Protokoll für Insektizide gegen Bettwanzen Eier mit einem Vergleich zum ersten nymphal Bett Bug Bühne zu bewerten. Diese Protokolle könnten für andere Insekten angepasst werden, um Insektizid Wirksamkeit bei Non-Adult Lebensphasen zu bewerten.

Ein Verfahren zur Bewertung Insecticide Wirksamkeit gegen Bettwanzen,<em&gt; Cimex lectularius</em&gt;, Eier und First Instars

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

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

11.2K Aufrufe.  University of Florida.  Insecticide Evaluierungen werden häufig gegen erwachsenen Insekten gezielt, anstatt unreifen Stadien. Hier präsentieren wir ein Protokoll für Insektizide gegen Bettwanzen Eier mit einem Vergleich zum ersten nymphal Bett Bug Bühne zu bewerten. Diese Protokolle könnten für andere Insekten angepasst werden, um Insektizid Wirksamkeit bei Non-Adult Lebensphasen zu bewerten. 

Serie: 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.

Eine schnelle und effiziente Methode zur Beurteilung der Pathogenität von Ustilago-Maydis auf Mais- und Teosintenlinien

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

Forschung

Umwelt

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.

Eine einfache Methode für die automatisierte Festphasenextraktion von Wasserproben für die immunologische Analyse von kleinen Schadstoffe

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.

Die Verwendung einer Batterie von chemischen und ökotoxikologischen Methoden zur Beurteilung der Wirksamkeit von Abwasserbehandlungsprozesse zu entfernen Estrogenic Potency

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.

Ein Gusseted Thermogradient Tabelle zur Bekämpfung von Bodentemperaturen für das Pflanzenwachstum Beurteilung und Überwachung von Bodenprozessen

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.

Zuverlässige Methode zur Beurteilung der Samenkeimung Dormanz und Mortalität unter Feldbedingungen

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.

Ein Durchflussbelichtungssystem für die Bewertung Schwebstoffe Auswirkungen auf Leben im Wasser

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.

Eine Lipid-Extraktions- und Analysemethode zur Charakterisierung von Boden-Mikroben in Experimenten mit vielen Proben

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.

Bildbasierte Lagrange-Partikel-Tracking in Bettlasten-Experimenten

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).

Eine neue Methode für die Analyse von Pentosan in Jute Biomasse und deren Umwandlung in Zucker Monomere mit sauren ionischen Flüssigkeit

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.

Wirksamkeit von Pestiziden gegen Krankheit Vektor Mücken im Bereich Visualisierung

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