This work is part of a CICYT research project (AGL 2010-22178) of the Spanish Ministry of Science and Technology. The authors are grateful to Nic Aldam and Kristi Prunty for the English revision of the manuscript.

Das Protokoll unten beschrieben für die Induktion CTA in Richtung Gehölzen folgt die Tierpflege Richtlinien der &quot;Universitat Autònoma de Barcelona&quot; (Bellaterra, Spanien) und wird von der Ethikkommission der Tier- und Menschenversuche für Schafe und Ziegen (CEEAH, verweist auf 770 genehmigt und 998 beziehungsweise). 1. Herstellung von LiCl Dose <ol><li> Machen Sie eine 25% w / v (Gewicht / Volumen) LiCl-Lösung in destilliertem Wasser. Alternativ verwenden Sie sauberes Leitungswasser. Die LiCl ist extrem hygroskopisch deshalb ist es notwendig, darauf zu achten, wenn das pulverförmige chemische Behandlung. <ol><li> Wiegen 250 g LiCl und löse es in destilliertem Wasser (LiCl Konzentratlösung). Da die Reaktion exotherm ist, warten Sie, bis die Lösung auf Raumtemperatur, bevor Sie fortfahren zurückgekehrt ist. Füllen Sie die Lösung in einen Messkolben mit destilliertem Wasser, das es bis 1000 ml zu bringen. </li></ol></li><li> Berechnen des Volumens der Lösung für jedes Tier (EinzelDosis). <ol><li> Wiegen Sie die Tiere ihr Körpergewicht (BW) zu erhalten , und berechnen die ml LiCl für jedes Tier benötigt folgende Gleichung 1. Die empfohlene Dosis für die Langzeit - Persistenz 225 mg LiCl / kg KG bei Schafen 10 und 200 mg LiCl / kg KG bei Ziegen 17. Jedoch können auch andere Dosen beweidet werden , sind in Tabelle 1 gezeigt. (Kg BW x LiCl Dosis in g / kg KG) / LiCl-Konzentration in g / l = Volumen des LiCl (L) [Gl. 1] Beispielsweise: Schaf = 60 kg BW; Dosis = 225 mg LiCl / kg KG; Konzentration von LiCl-Lösung = 250 g / L (60 kg x 0,225 g / kg) / 250 g / L = 0,054 L = 54 ml Ziege = 35 kg BW; Dosis = 200 mg LiCl / kg KG; Konzentration von LiCl-Lösung = 250 g / L (35 kg x 0,200 g / kg) / 250 g / L = 0,028 L = 28 ml </li></ol></li></ol><table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> LiCl Dosis </td><td> Mazorra et al. (2006) 19 </td&gt; <td> Burritt et al. (2013) 35 </td></tr><tr><td> Niedrig </td><td> 100 </td><td> 125 </td></tr><tr><td> Mittel </td><td> 150 </td><td> 150 </td></tr><tr><td> Hoch </td><td> 225 </td><td> 175 </td></tr></tbody></table> Tabelle 1:. LiCl Dosisbereich Bereich LiCl Dosen (mg / kg Körpergewicht, BW) von verschiedenen Autoren verwendet Futter Abneigung zu induzieren. 2. Tierauswahl und Ausstattung <ol><li> Wählen Sie für Erwachsene, nicht schwanger und trocken Tiere, die zuvor noch nie das Ziel Futter gefressen. </li><li> Nicht zur gleichen Zeit, um die CTA, um mehr als 20 Tieren induzieren, um besser das Verhalten steuern. Zunächst induzieren nur CTA zu 5 Tieren. </li><li> Ordnen Sie die Tiere in einem Ort, an dem die Aufnahme des Ziel Feed kann einzeln erfasst werden. Füttern Sie die Tiere einmal pro Tag mit einer Basal-Diät, die alle ihren Nährstoffbedarf und bieten freien Zugang zu Wasser und zu einem miNeral Block. Assure eine ausreichende Salzzufuhr LiCl toxische Effekte zu verhindern, da die Körperzellen Li anstelle von Na 9 verwenden können. </li><li> Wenn die Einrichtungen für die Tiere neu sind, geben sie von 1 Woche Anpassungszeit mit der Umgebung vertraut zu machen und die Grundnahrung. </li></ol> 3. Aversion Induction <ol><li> Entfernen Sie die Basal - Diät , bevor der Abend (Tag -1) und bieten die Ziel Feed ad libitum , um sicherzustellen , dass die Tiere 12 das Ziel - Feed eifrig am nächsten Tag essen. </li><li> Am folgenden Tag (Tag 0), die orts am Morgen entfernen und während 30 Minuten auf jedes Tier 200 g Ziel Futter bieten. Wiegen Sie die orts danach. </li><li> Wenn die Tiere mehr als 20-30 g verbrauchen, zur Verwaltung der entsprechenden berechneten Volumen von LiCl (ml berechnet in 1.2) mit einer Durchnässung Pistole so bald wie praktisch möglich ist, nicht mehr als 1 Stunde nach dem Verzehr warten. Wenn Tiere verbrauchen weniger als 20-30 g, wiederholen Sie den Vorgang am nächsten Tag. Due zu ihrer angeborenen neophobic Einzugsverhalten, Tiere könnten mehr als 1 Tag müssen die Ziel Feed 10,17,25-27 beginnen verbrauchen. </li><li> Nach LiCl Verwaltung, warten 2 Stunden vor der Basal-Diät anbietet. </li><li> Von Tag 1 bis 3, bieten nicht die Tiere, die die Ziel Futter und überprüfen sie regelmäßig alle möglichen Zeichen einer schweren Krankheit zu erkennen, tägliche Futter- und Wasseraufnahme (nach Gewicht oder durch eine Check-Liste), Atemfrequenz und das Verhalten der Tiere aufnehmen. Sie könnten präsentieren fiel Kopf und Ohren, Inaktivität, Durchfall, erhöhte Atemfrequenz und verringert Wasser und Nahrungsaufnahme in den Tagen nach der LiCl Verwaltung 9-11,20,25. </li></ol> 4. Aversion Validation <ol><li> An den Tagen, 4, 5, 6 und 7 nach LiCl Verwaltung, wiederholen Sie den Schritt 3.2. </li><li> Wenn die Tiere verbrauchen &lt;10 g (oder &lt;4 Bisse) der Ziel Futtermittel, betrachten die CTA established.The Schwelle zwischen Null (kein Verbrauch) und 10 g auf den Umstand abhängen könnte vondie Zielpflanze (toxische oder nicht). </li><li> Wenn ein Tier verbraucht&gt; 10 g, verwalten eine neue LiCl Dosis und wiederholen Sie Schritt 4.1. Wenn das Tier in Konsum der Zielpflanze nach der zweiten Dosis bestehen, beseitigen sie von der CTA-Gruppe. Soziale Erleichterung (zB Mutter, Geschwister und enge Kumpels) können CTA Verhalten im Tierbereich ändern. Tiere , die zu einer Zielpflanze abgewendet wurden leicht verbrauchen sie wieder , wenn sie mit nicht abgewendeten Tiere grasen 28. </li></ol> 5. Weidemanagement Anmerkung: Die Dosis von LiCl verwendet CTA zu induzieren vollständig in wenigen Tagen ausgeschieden wird, hauptsächlich durch Urin. <ol><li> Obwohl Li weit auf der Erdkruste verteilt wird, um eine mögliche Kontamination in der organischen Ernte zu vermeiden, warten Sie 9-11 Tage vor , die Tiere der Ernte (Clearance - Zeit etabliert mit Li pharmakokinetischen) 11 bewegt. </li><li> Stellen Sie sicher, dass die Ernte Bodendecke ist reichlich vorhanden und schmackhaft im ganzendie Tiere zu verhindern, dass das Abtasten des Zielzuteilzeit weiden lassen. Graze behandelten Tiere getrennt von nicht behandelten. </li><li> Lassen Sie die Tiere ein delimitiert Grundstück von der gesamten Oberfläche zu grasen eine einheitliche Steuerung der Grashöhe zu erhalten. Abgrenzen die Handlung von tragbaren elektrischen oder metallischen Zäunen verwendet oder einen Schäfer mit den Tieren zwischen spezifischen Pflanzenlinien zu grasen. </li><li> Vermeiden Sie ein Grundstück Überweidung, Beseitigung der Tiere , bevor die Bodenabdeckung knapp wird (Grashöhe von 5 cm) 29. </li></ol> 6. Wiedereinführung der CTA <ol><li> Jedes Jahr, bevor die Tiere erlaubt zurückzukehren, um die Ernte zu grasen, wiederholen Sie Schritt 4. </li></ol>

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The authors have nothing to disclose.

Das CTA ist leicht bei kleinen Wiederkäuern festgelegt, wenn das Ziel-Feed ist eine Pflanze, die das Tier noch nie gegessen hat und nicht eine unersetzliche Nährstoff enthalten. Tiere haben eine positive post-Ingestiv Verbindung mit einem nicht-toxischen Futter , es sei denn vorherigen Kontakt macht es schwierig , ihre Wahrnehmung dieser besonderen Futter 7,33 zu ändern. Die Bedingung Geschmacksaversion wird produziert , weil LiCl das Brechmittel - System produzieren Unwohlsein oder Magen - Darm - Beschwer...

Der Einsatz von Bodenbedeckung zwischen holzigen Pflanzenlinien mildert Bodenerosion und Abbau und erhöht Wasser, organischem Kohlenstoff und Stickstoff - Retention 1-3. Darüber hinaus unterhält Bodenabdeckung und erhöht die biologische Vielfalt, die das Gleichgewicht zwischen den Pflanzenschädlinge und ihre natürlichen Feinde zu unterstützen. Die Landwirte sind in der Regel Unkraut zu beseitigen, indem agrochemischen Produkten Anwendung oder eine Mähmaschine Maschine; wodurch Nährstoffkonkurrenz zwischen den Kulturen und Bewuchs. Eine kostengünstige Möglichkeit, Bodendecker zu steuern würde die Verwendung von kleinen Wiederkäuern Weide sein. Ein weiterer Vorteil von Tier Beweidung ist die Verbesserung der Bodengesundheit und Fruchtbarkeit. Allerdings sind die Landwirte zögern, diese Praxis aufgrund von kleinen Wiederkäuern durch den Verzehr von jungen Blättern und Sprossen zu schädigen die Pflanzen zu implementieren. Um Schäden zu vermeiden potenzielle Ernte ist es nützlich, konditionierten Geschmacksaversion (CTA) in den Schafen oder Ziegen in der Herde oder Herde zu induzieren. Das CTA ist leicht für n etabliertew - Feeds, aufgrund der angeborenen Futter neophobia Verhalten 4,5 von kleinen Wiederkäuern, und weil vertraute Feeds positiv mit einem &quot;Erlernte Sicherheit&quot; Status zugeordnet sind , die schwieriger zu ändern oder 6 manipulieren. Tiere lernen, einen bestimmten Feed (bedingten Reiz) aufgrund seiner negativen postingestive Wirkung (unbedingter Reiz) abzulehnen. Zu induzieren CTA Richtung schmackhafter und ungiftig Pflanzen, Lithiumchlorid (LiCl; Induktionsmittel) oral verabreicht wird, nachdem das Tier die Zielpflanze verbraucht. Während es andere Induktor Mittel (zB Apomorphin, ciclosphosphamide, Thiabendazol) sind, zeigten LiCl die stärkste und persistent CTA aufgrund seiner Wirkung auf das Brechmittel - System durch Stimulation der Chemorezeptor - Triggerzone Bereich und gastrointestinale Beschwerden 7,8 mit milden Anzeichen der allgemeinen Beschwerden. Lithium (Li) aus dem oberen Magen - Darm - Trakt resorbiert und in das Gesamtkörperwasser Raum 9 verteilt. Die Tiere can eine Erholungsphase so kurz wie 2 Tage 7,10,11. Der LiCl kann es durch Mischen mit dem Lebensmittel 12,13 oral 15-17 durch eine Durchnässung Pistole verabreicht in einer Gelatinekapsel 13,14 oder in einer Lösung verabreicht werden. Obwohl LiCl Lösung ätzend ist, wurde keine Verletzung im Mund oder der Speiseröhre beschrieben. LiCl wird im Bereich von 100 bis 400 mg LiCl / kg Körpergewicht (BW), mit besseren Ergebnissen (hartnäckiger CTA) mit höheren Dosen 16,18 verwendet. Dennoch, die bekannten Dosierung Auswirkungen auf verschiedene Arten und Rassen unter Berücksichtigung, die letale Wirkung in einigen Fällen beginnen bei 400 mg LiCl / kg KG. Die empfohlene Dosierung für eine effektive langfristige CTA beginnt bei 200 mg / kg KG für Ziegen und 225 mg / kg KG für Schafe 10,17,19. Li bei diesen Dosierungen innerhalb der ersten 4 Tage nach der Verabreichung ausgeschieden wird, hauptsächlich durch den Urin (92 ± 4%), gefolgt von Fäzes (6,5 ± 1,3%) und Milch (2,8 ± 0,4%) 11. Das Ganzegeschätzte Wartezeit für eine Einzeldosis von LiCl in Plasma 9 und 11 Tagen bei Schafen und Ziegen sind. Aufgrund der geringen Li - Ausscheidung in der Milch, kann CTA nicht natürlich off-Frühling 11,20 in den gestillten hergestellt werden. Die langfristigen CTA Persistenz bei Schafen wurde während einer gesamten Weidesaison (3-4 Monate) , wenn eine alternative Futterquelle 14,21 verfügbar war berichtet, mit einer einzigen Dosis LiCl bei der nächsten Beweidung zu einer fast vollständigen Aversion wieder hergestellt werden Saison (9 Monate später) 14. Darüber hinaus CTA persistences von 2 und 3 Jahren in Kühe unter Weidebedingungen, ohne die Notwendigkeit einer Verstärkung Dosen, wenn das Ziel - Feed ist eine giftige , aber schmackhaft Pflanze 22,23 berichtet. Die Möglichkeit, eine alternative Futtermittel unter Berücksichtigung ist von entscheidender Bedeutung für das Tier das CTA gegen einen nicht-toxischen Pflanze zu erhalten. Jedesmal, wenn das Tier verbraucht mehr als 10 g des abgewendet Anlage ohne resultierende in gastrointestinalen discomfort würde die CTA 24 beeinträchtigt werden.

<table border="0" cellpadding="0" cellspacing="0" width="598">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Lithium Chloride PRS</td>
			<td style="width:71px;">Panreac</td>
			<td align="right" style="width:119px;">141392.1209</td>
			<td style="width:192px;">Different amounts of same product can be supplied by the same company.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Labelvage drencher 70 mL</td>
			<td style="width:71px;">Labelvage</td>
			<td align="right" style="width:119px;">240040</td>
			<td style="width:192px;">Similar product can be used (different brand or volume).</td>
		</tr>
	</tbody>
</table>

how to create conditioned taste aversion for grazing ground covers in woody crops with small ruminants

Conditioned taste aversion (CTA) is a learning behavior process where animals are trained to reject certain feed after gastrointestinal discomfort has been produced. Lithium chloride (LiCl) is the preferred agent used in livestock to induce CTA because it specifically stimulates the vomit center. In addition, LiCl is commercially available, and easy to prepare and administer using a drenching gun. Nevertheless, some factors have to be considered to obtain an effective long-lasting CTA, which allows small ruminants to graze during the cropping season. A key aspect is to use animals with no previous contact with the target plant (the plant chosen to be avoided; new feed). Due to their native neophobic feeding behavior, small ruminants can easily associate the negative feedback effects with the new feed, resulting in a strong and persistent CTA. The recommended doses are 200 and 225 mg LiCl/kg body weight (BW) for goats and sheep, respectively. To induce CTA, 100 g of the target plant should be individually offered for at least 30 min, and LiCl administered thereafter if the intake is greater than 10 g. Each time the animal eats the target plant without negative consequences, the CTA becomes weaker. Consequently, to minimize the risk of target plant consumption, it is essential to have sufficient palatable ground cover available. The presence of an alternative feed (of quality and quantity) prevents the accidental consumption of the target plant. A close monitoring of the flock is recommended to remove and re-dose any animal consuming more than 4 bites or 10 g of the target plant. At the beginning of each grazing season, check the CTA status of each animal before moving them to the crop.

Die im Folgenden beschriebenen Ergebnisse wurden durch mehrere Studien in CTA zu Gehölzen bei kleinen Wiederkäuern an der Universitat Autònoma de Barcelona durchgeführt erhalten und sie den Nachweis erbringen, das Protokoll vorgeschlagen zu etablieren. Dosen von 175 und 200 mg LiCl / kg KG bei Ziegen und 200 und 225 mg LiCl / kg KG bei Schafen wurden erfolgreich eingesetzt CTA zu induzieren gegen Gehölzen mit einer einzig...

Watch this Scientific Journal Video about How to Create Conditioned Taste Aversion for Grazing Ground Covers in Woody Crops with Small Ruminants at JoVE.com

How to Create Conditioned Taste Aversion for Grazing Ground Covers in Woody Crops with Small Ruminants

Wie konditionierten Geschmacksaversion zu erstellen für die Beweidung Bodendecker in Gehölzen mit kleinen Wiederkäuer

Feeding behavior preferences in livestock can be modified to implement a grazing management plan in woody crops. Here, we present a protocol to show a lithium chloride dose after eating a new plant that induces conditioned taste aversion.

Conditioned taste aversion (CTA) is a learning behavior process where animals are trained to reject certain feed after gastrointestinal discomfort has ...

how-to-create-conditioned-taste-aversion-for-grazing-ground-covers

Research

JoVE Journal

Environment

7.2K Aufrufe.  Universitat Autònoma de Barcelona. Feeding behavior preferences in livestock can be modified to implement a grazing management plan in woody crops. Here, we present a protocol to show a lithium chloride dose after eating a new plant that induces conditioned taste aversion. 

Serie: How to Create Conditioned Taste Aversion for Grazing Ground Covers in Woody Crops with Small Ruminants

Surface Renewal: An Advanced Micrometeorological Method for Measuring and Processing Field-Scale Energy Flux Density Data

Advanced micrometeorological methods have become increasingly important in soil, crop, and environmental sciences. For many scientists without formal training in atmospheric science, these techniques are relatively inaccessible. Surface renewal and other flux measurement methods require an understanding of boundary layer meteorology and extensive training in instrumentation and multiple data management programs. To improve accessibility of these techniques, we describe the underlying theory of surface renewal measurements, demonstrate how to set up a field station for surface renewal with eddy covariance calibration, and utilize our open-source turnkey data logger program to perform flux data acquisition and processing. The new turnkey program returns to the user a simple data table with the corrected fluxes and quality control parameters, and eliminates the need for researchers to shuttle between multiple processing programs to obtain the final flux data. An example of data generated from these measurements demonstrates how crop water use is measured with this technique. The output information is useful to growers for making irrigation decisions in a variety of agricultural ecosystems. These stations are currently deployed in numerous field experiments by researchers in our group and the California Department of Water Resources in the following crops: rice, wine and raisin grape vineyards, alfalfa, almond, walnut, peach, lemon, avocado, and corn.

Surface renewal is a micrometeorological method that is being used increasingly to determine energy fluxes, but its technical complexity makes it inaccessible to a broad audience. We describe the steps needed to set up and calibrate a surface renewal field station, to acquire and process data, and to correctly interpret results.

Oberflächen Erneuerung: Ein Advanced Mikrometeorologische Verfahren zur Messung und Verarbeitung Feld Skala Energieflussdichte Daten

Advanced micrometeorological methods have become  increasingly important in soil, crop, and environmental sciences.  For many scientists without formal ...

Forschung

Umwelt

A Small Volume Procedure for Viral Concentration from Water

Small-scale concentration of viruses (sample volumes 1-10 L, here simulated with spiked 100 ml water samples) is an efficient, cost-effective way to identify optimal parameters for virus concentration. Viruses can be concentrated from water using filtration (electropositive, electronegative, glass wool or size exclusion), followed by secondary concentration with beef extract to release viruses from filter surfaces, and finally tertiary concentration resulting in a 5-30 ml volume virus concentrate. In order to identify optimal concentration procedures, two different electropositive filters were evaluated (a glass/cellulose filter [1MDS] and a nano-alumina/glass filter [NanoCeram]), as well as different secondary concentration techniques; the celite technique where three different celite particle sizes were evaluated (fine, medium and large) followed by comparing this technique with that of the established organic flocculation method. Various elution additives were also evaluated for their ability to enhance the release of adenovirus (AdV) particles from filter surfaces. Fine particle celite recovered similar levels of AdV40 and 41 to that of the established organic flocculation method when viral spikes were added during secondary concentration. The glass/cellulose filter recovered higher levels of both, AdV40 and 41, compared to that of a nano-alumina/glass fiber filter. Although not statistically significant, the addition of 0.1% sodium polyphosphate amended beef extract eluant recovered 10% more AdV particles compared to unamended beef extract.

An approach was developed for identifying optimal viral concentration conditions for small volume water samples using spikes of human adenovirus. The techniques described here are used to identify concentration parameters for other viral targets, and applied to large-scale viral concentration experimentation.

Ein kleines Volumen Verfahren für Viral Konzentration von Wasser

Small-scale concentration of viruses (sample volumes 1-10 L, here simulated with spiked 100 ml water samples) is an efficient, cost-effective way to ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. The standardized sampling component of the method concentrates viruses that may be present in water by passage of a minimum specified volume of water through an electropositive cartridge filter. The minimum specified volumes for surface and finished/ground water are 300 L and 1,500 L, respectively. A major method limitation is the tendency for the filters to clog before meeting the sample volume requirement. Studies using two different, but equivalent, cartridge filter options showed that filter clogging was a problem with 10% of the samples with one of the filter types compared to 6% with the other filter type. Clogging tends to increase with turbidity, but cannot be predicted based on turbidity measurements only. From a cost standpoint one of the filter options is preferable over the other, but the water quality and experience with the water system to be sampled should be taken into consideration in making filter selections.

EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.

EPA-Methode 1615 Messung der Enterovirus und Norovirus Vorkommen in Wasser von Kultur und RT-qPCR. I. Sammlung von Virenproben

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

A Technical Perspective in Modern Tree-ring Research - How to Overcome Dendroecological and Wood Anatomical Challenges

Dendroecological research uses information stored in tree rings to understand how single trees and even entire forest ecosystems responded to environmental changes and to finally reconstruct such changes. This is done by analyzing growth variations back in time and correlating various plant-specific parameters to (for example) temperature records. Integrating wood anatomical parameters in these analyses would strengthen reconstructions, even down to intra-annual resolution. We therefore present a protocol on how to sample, prepare, and analyze wooden specimen for common macroscopic analyses, but also for subsequent microscopic analyses. Furthermore we introduce a potential solution for analyzing digital images generated from common small and large specimens to support time-series analyses. The protocol presents the basic steps as they currently can be used. Beyond this, there is an ongoing need for the improvement of existing techniques, and development of new techniques, to record and quantify past and ongoing environmental processes. Traditional wood anatomical research needs to be expanded to include ecological information to this field of research. This would support dendro-scientists who intend to analyze new parameters and develop new methodologies to understand the short and long term effects of specific environmental factors on the anatomy of woody plants.

Here we present a protocol outlining how to sample wooden specimens for the overall assessment of their growth structures. Macro- and microscopic preparation and visualization techniques necessary to generate well-replicated and highly resolved wood anatomical and dendroecological dataset, are described are described.

Technischer Sicht in Modern Jahrringforschung - Wie Dendroökologische und Holz anatomische Herausforderungen zu meistern

Dendroecological research uses information stored in tree rings to understand how single trees and even entire forest ecosystems responded to ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a standardized method for use in multiple analytical laboratories during monitoring period 3 of the Unregulated Contaminant Monitoring Rule. Herein we present the protocol for extraction of viral ribonucleic acid (RNA) from water sample concentrates and for quantitatively measuring enterovirus and norovirus concentrations using reverse transcription-quantitative PCR (RT-qPCR). Virus concentrations for the molecular assay are calculated in terms of genomic copies of viral RNA per liter based upon a standard curve. The method uses a number of quality controls to increase data quality and to reduce interlaboratory and intralaboratory variation. The method has been evaluated by examining virus recovery from ground and reagent grade waters seeded with poliovirus type 3 and murine norovirus as a surrogate for human noroviruses. Mean poliovirus recoveries were 20% in groundwaters and 44% in reagent grade water. Mean murine norovirus recoveries with the RT-qPCR assay were 30% in groundwaters and 4% in reagent grade water.

Here we present a procedure to quantify enterovirus and norovirus in environmental and drinking waters using reverse transcription-quantitative PCR. Mean virus recovery from groundwater with this standardized procedure from EPA Method 1615 was 20% for poliovirus and 30% for murine norovirus.

EPA-Methode 1615 Messung der Enterovirus und Norovirus Vorkommen in Wasser, Kultur und RT-qPCR. Teil III. Virenerkennung durch RT-qPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a ...

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 ...

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in nutrient and gas cycles. After cores with intact sediment-water interfaces are collected, they are submerged in incubation tanks and kept under aerobic conditions at in situ temperatures. To initiate a time course of overlying water chemistry, cores are sealed without bubbles using a top cap with a suspended stirrer. Time courses of 4-7 sample points are used to determine the rate of sediment water exchange. Artificial illumination simulates day-time conditions for shallow photosynthetic sediments, and in conjunction with dark incubations can provide net exchanges on a daily basis. The net measurement of N2 is made possible by sampling a time course of dissolved gas concentrations, with high precision mass spectrometric analysis of N2:Ar ratios providing a means to measure N2 concentrations. We have successfully applied this approach to lakes, reservoirs, estuaries, wetlands and storm water ponds, and with care, this approach provides valuable information on biogeochemical balances in aquatic ecosystems.

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

Here, we present small core incubations for the measurement of sediment-water gas and solute exchange. These will provide reliable measurements of sediment-water exchange that assess the role of sediment in influencing biological and biogeochemical processes in aquatic ecosystems.

Die Benthic Austausch von O<sub&gt; 2</sub&gt;, N<sub&gt; 2</sub&gt; Und gelöste Nährstoffe mit kleinen Kern Inkubationen

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in ...

Tracking Infiltration Front Depth Using Time-lapse Multi-offset Gathers Collected with Array Antenna Ground Penetrating Radar

A Ground Penetrating Radar (GPR) system based on a ground-coupled, densely populated antenna array was used to collect data during an infiltration experiment conducted at a test site near the Tottori Sand Dune, Japan. The antenna array used in this study consists of 10 transmitting antennas (Tx) and 11 receiving antennas (Rx). For this experiment, the system was configured to use all possible Tx-Rx pairings, resulting in a Multi-Offset Gather (MOG) consisting of 110 Tx-Rx combinations. The array was left stationary at a position directly above the infiltration area and data were collected every 1.5 seconds using a time-based trigger. Common-Offset Gather (COG) and Common Mid-Point (CMP) data cubes were reconstructed from the MOG data during post-processing. There have been few studies that used time-lapse CMP data to estimate changes in velocity of propagation. In this study, electromagnetic (EM) wave velocity was estimated heuristically at 1-minute intervals from the reconstructed CMP data through curve fitting, using the hyperbola equation. We then proceeded to calculate the depth of the wetting front. The evolution of the wetting front over time obtain through this method is consistent with the observations from a soil moisture sensor which was placed at a depth below 20 cm. The results obtained in this study demonstrate the ability of such array GPR system to monitor a subsurface dynamic process like water infiltration accurately and quantitatively.

Here we present a Ground Penetrating Radar (GPR) system based on a ground-coupled, densely populated antenna array for monitoring the dynamic process of subsurface water infiltration. A time-lapse radar image of the infiltration process allowed estimating the depth of the wetting front during the course of the infiltration process.

Tracking-Infiltration, die vorderen Tiefe mit Zeitraffer Multi-Offset sammelt mit Array Antenne Bodenradars gesammelt

A Ground Penetrating Radar (GPR) system based on a ground-coupled, densely populated antenna array was used to collect data during an infiltration ...

Experimental Methods of Dust Charging and Mobilization on Surfaces with Exposure to Ultraviolet Radiation or Plasmas

Electrostatic dust transport has been hypothesized to explain a number of observations of unusual planetary phenomena. Here, it is demonstrated using three recently developed experiments in which dust particles are exposed to thermal plasma with beam electrons, beam electrons only, or ultraviolet (UV) radiation only. The UV light source has a narrow bandwidth in wavelength centered at 172 nm. The beam electrons with the energy of 120 eV are created with a negatively biased hot filament. When the vacuum chamber is filled with the argon gas, a thermal plasma is created in addition to the electron beam. Insulating dust particles of a few tens of microns in diameter are used in the experiments. Dust particles are recorded to be lofted to a height up to a few centimeters with a launch speed up to 1 m/s. These experiments demonstrate that photo and/or secondary electron emission from a dusty surface changes the charging mechanism of dust particles. According to the recently developed &#34;patched charge model&#34;, the emitted electrons can be re-absorbed inside microcavities between neighboring dust particles below the surface, causing the accumulation of enhanced negative charges on the surrounding dust particles. The repulsive forces between these negatively charged particles may be large enough to mobilize and lift them off the surface. These experiments present the advanced understanding of dust charging and transport on dusty surfaces, and laid a foundation for future investigations of its role in the surface evolution of airless planetary bodies.

Dust charging and mobilization is demonstrated in three experiments with exposure to thermal plasma with beam electrons, beam electrons only, or ultraviolet (UV) radiation only. These experiments present the advanced understanding of electrostatic dust transport and its role in shaping the surfaces of airless planetary bodies.

Experimentelle Methoden der Aufladung Staub und Mobilisierung auf Oberflächen mit der Exposition gegenüber ultravioletter Strahlung oder Plasmen

Electrostatic dust transport has been hypothesized to explain a number of observations of unusual planetary phenomena. Here, it is demonstrated using ...

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current measures to combat such pathogens have proved too conservative and, thus, not sufficiently effective, and they can potentially pose environmental risks. Therefore, it is extremely necessary to identify host-resistance factors to assist in controlling plant diseases naturally through the identification of resistant germplasm, the isolation and characterization of resistance genes, and the molecular breeding of resistant cultivars. In this regard, there is need to establish an accurate, rapid, and large-scale inoculation method to breed and develop plant resistance genes. The rice blast fungal pathogen Magnaporthe grisea causes severe disease symptoms and yield losses. Recently, M. grisea has emerged as a model organism for studying the mechanisms of plant-fungal pathogen interactions. Hence, we report the development of a plant virulence test method that is specific for M. grisea. This method provides for both spray inoculation with a conidial suspension and wounding inoculation with mycelium cubes or droplets of conidial suspension. The key step of the wounding inoculation method for detached rice leaves is to make wounds on plant leaves, which avoids any interference caused by host penetration resistance. This spray/wounding protocol contributes to the rapid, accurate, and large-scale screening of the pathotypes of M. grisea isolates. This integrated and systematic plant infection method will serve as an excellent starting point for gaining a broad perspective of issues in plant pathology.

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Here, we present a protocol to test plant virulence with the plant pathogen Magnaporthe grisea. This report will contribute to the large-scale screening of the pathotypes of fungi isolates and serve as an excellent starting point for understanding the resistant mechanisms of plants during molecular breeding.

Die Pflanze-Infektion-Test: Spray und Wunde-vermittelten Inokulation mit Pflanzen Pathogen Magnaporthe Grisea

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current ...