Wir danken Manel Bolivar für seine Unterstützung bei der Feldarbeit. Wir sind dankbar, dass der &quot;Parc Natural del Montgrí, les Illes Medes i el Baix Ter&quot; für ihre Unterstützung unserer Forschungs- und Sampling-Berechtigungen. Die Unterwasser-Manipulator wurde von Ayelet Dadon-Pilosof entworfen und hergestellt von Herrn Pilosof. Diese Arbeit wurde von der spanischen Regierung Projekt CSI-Coral [Gewährungsnummer CGL2013-43106-R RC und MR] und durch eine FPU Stipendium &quot;Ministerio de Educación, Cultura y Deporte (MECD)&quot; auf TM unterstützt. Dies ist ein Beitrag aus dem Marine Biogeochemie und Global Change Forschungsgruppe von der katalanischen Regierung [Grant-Nummer 2014SGR1029] und ISF Zuschuss 1280/13 und BSF Zuschuss 2.012.089 G. Yahel finanziert.

<ol>
	<li>Gili, J. M., Coma, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Benthic+suspension+feeders:+their+paramount+role+in+littoral+marine+food+webs.">Benthic suspension feeders: their paramount role in littoral marine food webs.</a> Trends. Ecol. Evol. 13 (8), 316-321 (1998).</li><li>Reiswig, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+situ+pumping+activities+of+tropical+Demospongiae.">In situ pumping activities of tropical Demospongiae.</a> Mar. 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The authors have nothing to disclose.

Vorbereitende Schritte Collector Fläschchen für DOM und Nährstoffanalysen Da Kollektor Gefäße können mit gelösten Mikrokomponenten interagieren und die Sampler Wände 30-34, verschiedene Fläschchen für DOM und Nährstoff Sammlung wurden getestet kann ein Substrat für das Wachstum von Bakterien. Borosilikat ist nicht für die Kieselsäure Quantifizierung empfohlen 33,35, da Glasflaschen die...

Benthic Filtrierer spielen eine entscheidende Rolle in der Funktionsweise der marinen Ökosysteme 1. Durch das Filtern von großen Mengen Wasser 2,3, sie zu entfernen und Partikel (Plankton und Geröll) und gelösten Verbindungen 1 (und Referenzen darin) und sind ein wichtiger Agent der Benthos-pelagische Kopplung 4,5 und Nährstoffkreislauf 6,7 ausscheiden. Genau das Partikelmess und gelösten Verbindungen entfernt und durch benthische Filtrierer (wie Schwämme, Seescheiden, Polychaeten und bivalves) ausgeschieden ist von grundlegender Bedeutung, ihre Physiologie, Stoffwechsel zu verstehen und Nahrungsökologie. Zusammen mit Geschwindigkeitsmessungen Pumpen, sondern ermöglicht auch eine Quantifizierung der Nährstoffflüsse durch diese Organismen und ihre ökologischen Auswirkungen auf die Wasserqualität sowie auf Ökosystemebene Prozesse vermittelt. Die Wahl der geeigneten Methode zur Messung der Entfernung und Produktionsraten von partikulären und gelösten comPfund durch Suspension Filtrierer ist entscheidend für die zuverlässige Daten zu erhalten , die ihre Fraßaktivität 8. Wie durch Riisgård und andere unangemessene Methoden Bias-Ergebnisse darauf hingewiesen, verzerren experimentellen Bedingungen zu falschen Einschätzungen der Einnahme und Ausscheidung bestimmter Stoffe, und von diesen Organismen verarbeitet, um fehlerhafte Quantifizierung der Nährstoffflüsse führen kann. Die beiden am häufigsten verwendeten Methoden partikulären und gelösten Nährstoffflüsse in Filtrierer beinhalten messen entweder Inkubation (indirekte Techniken) oder simultane Erfassung von Umgebungs- und atmete aus Wasser (direkte Techniken). Die Inkubation Techniken basieren in der Konzentration von partikulären und gelösten Nährstoffen im Wasser inkubiert, die Änderungsrate auf Messen und 8 Produktionsraten oder Entfernung im Vergleich zu angemessenen Kontrollen abzuschätzen. Jedoch kann ein Organismus in einer Inkubationskabine umschließt seine feedin änderng und Pumpverhalten aufgrund von Änderungen in der natürlichen Strömungsbereich aufgrund einer Abnahme der Sauerstoff und / oder in der Nahrungsmittelkonzentration oder aufgrund der Ansammlung von Ausscheidungsverbindungen in der Inkubation 7,9 Wasser (und Referenzen darin). Zusätzlich zu den Auswirkungen der Confinement und modifizierte Wasserversorgung stammt ein großer Vorspannung der Inkubation Techniken aus re Filtrationswirkungen (siehe beispielsweise 10). Obwohl einige dieser methodischen Probleme haben mit der rechten Form und Volumen des Inkubationsgefäßes 11 oder mit der Einführung einer Umlaufglasglocke System in situ 12, diese Technik oft unterschätzt Entfernung und Produktionsraten überwunden. Quantifizieren des Metabolismus von gelösten Verbindungen, wie gelösten organischen Stickstoff (DON) und Kohlenstoff (DOC) oder anorganische Nährstoffe, hat sich als besonders zu sein , anfällig für Verzerrungen verursacht durch Inkubation Techniken 13. In den späten 60er und frühen 70er Jahre, Henry Reiswig9,14,15 Vorreiter bei der Anwendung der direkten Techniken zur Partikelentfernung von riesigen Karibik Schwämme durch getrenntes Abtasten des Wasser ein- und ausgeatmeten durch die Organismen in situ, zu quantifizieren. Aufgrund der Schwierigkeit Reiswig der Technik , die auf kleinere Filtrierer und in anspruchsvoller Unterwasserbedingungen, der Großteil der Forschung auf diesem Gebiet an das Labor wurde anzuwenden beschränkt (in vitro) 16 meist indirekte Inkubationstechniken einsetzt. Yahel und Kollegen wieder montiert Reiswig direkte in situ - Technik in kleinerem Maßstab Bedingungen zu arbeiten. Ihre Methode, genannt 16 INEX, basiert auf der gleichzeitigen Unterwasserprobenentnahme des Wassers inhaliert (In) und abgeatmet (Ex) durch ungestörte Organismen. Die unterschiedliche Konzentration einer Substanz (zB Bakterien) zwischen einem Paar von Proben (inex) liefert ein Maß für die Retention (oder Produktion) dieses Stoffes durch das Tier. Die INEX Technik verwendet offene Röhren undberuht auf der excurrent Strahl durch die Pumpaktivität des untersuchten Organismus produziert, um das umgebende Wasser in das Sammelrohr passiv ersetzen. Während Yahel und Kollegen diese Technik erfolgreich bei der Untersuchung von mehr als 15 verschiedenen Aussetzung beantragt haben Beschicker Taxa (zB 17), wobei das Verfahren durch das hohe Maß an Übung und Erfahrung erforderlich, von der winzigen Größe einiger excurrent Öffnungen beschränkt ist, und durch Seegang. Um diese Hindernisse zu überwinden, entwickelten wir eine alternative Technik basiert auf kontrollierten Absaugung des abgetasteten Wasser durch winzige Röhrchen (Außendurchmesser &lt;1,6 mm). Unser Ziel war es, eine einfache, zuverlässige und kostengünstige Vorrichtung zu schaffen , die in situ Wasserentnahme aus einem ganz bestimmten Punkt, wie die excurrent Öffnung benthischer Filtrierer sauber und kontrolliert erlauben würde. Um wirksam zu sein, hat das Verfahren nicht aufdringlich sein, um nicht die Umgebungsströmungsregime zu beeinflussen oder die b ändernehavior der untersuchten Organismen. Das Gerät präsentiert hier wird VACUSIP bezeichnet. Es ist eine Vereinfachung des SIP - System entwickelt von Yahel et al. (2007) 18 für ROV-basierte Punktabtastung in der Tiefsee. Die VACUSIP ist wesentlich billiger als die ursprüngliche SIP und es hat für SCUBA-basierte Arbeit angepasst. Das System wurde nach den Prinzipien entworfen präsentiert und getestet von Wright und Stephens (1978) 19 und Møhlenberg und Riisgård (1978) 20 für Labor - Einstellungen. Obwohl das VACUSIP System für in - situ - Untersuchungen des Stoffwechsels von benthic Filtrierer entwickelt wurde, kann es auch für Laborstudien verwendet werden , und überall dort , wo eine kontrollierte und saubere, Point-Source - Wasser - Probe erforderlich ist. Das System ist besonders nützlich , wenn die Integration über längere Zeit (min-Stunden) oder in situ Filtrationen erforderlich sind. Die VACUSIP ist seit 2011 erfolgreich am Yahel Labor verwendet, und hat auchwurde in zwei aktuellen Studien von Nährstoffflüsse vermittelt durch Karibik und Mittelmeer Schwammarten 21 (Morganti et al. eingereicht) eingesetzt. Die Verwendung spezieller Sampler, der verlängerten Abtastdauer, und den Feldbedingungen, in denen VACUSIP angelegt wird, ziehen einige Abweichungen von Standardprotokollen ozeanographischer zum Sammeln, Filtern und Proben für empfindliche Analyte zu speichern. Um das Risiko einer Kontamination durch das VACUSIP System oder das Risiko einer Modifikation des beprobten Wasser durch bakterielle Aktivität nach der Entnahme zu verringern, haben wir getestet , verschiedene in situ Filtration und Lagerungsverfahren. Verschiedene Filtervorrichtungen, Sammelgefäße und Speichern Verfahren wurden untersucht , um die am besten geeignete Technik für die Analyse von gelösten anorganischen (PO 4 3-, NO x -, NH 4 +, SiO 4) zu erreichen , und organische (DOC + DON) Verbindungen und ultra Plankton (&lt;1081; m) und partikulärer organischer (POC + PON) Probenahme. Um weiter das Risiko einer Kontamination zu reduzieren, vor allem unter Freilandbedingungen wurde die Anzahl der Handhabungsschritte auf ein Minimum reduziert. Die visuelle Format, in dem das Verfahren dargestellt ist, ausgerichtet Reproduzierbarkeit zu erleichtern und um die Zeit zu reduzieren, die erforderlich, um effizient die Technik anzuwenden. Systemübersicht Zur in situ gepumpte Wasser aus der Suspension Abzweige mit exhalant Öffnungen so klein wie 2 mm, die Pumpaktivität jeder Probe Die Probe wird zuerst durch Lösen gefiltert Fluorescein gefärbt Meerwasser neben der inhalant Öffnung (en) sichtbar gemacht und das Beobachten seiner Strömung von der excurrent Öffnung 16 (siehe auch 2B Abbildung in 18). Das Wasser ein- und ausgeatmeten durch die Studie Probe (angefallenen und excurrent) werden dann mit der Verwendung eines Paares von kleinen Röhren installiert maßgeschneiderte Manipulator oder auf zwei der &quot;ar gleichzeitig abgetastetms &quot;eines upside-down flexible tragbare Stativ (Abbildung 1 und Ergänzungs Video 1). Das Wasser in der Studie Organismus inhaliert wird durch sorgfältiges Positionieren des proximalen Endes eines Rohres innerhalb oder in der Nähe der inhalant Apertur der Studie Organismus gesammelt. Eine identische Rohr dann innerhalb des excurrent Öffnung positioniert ist. Diese Operation gute Pflege erfordert Kontakt oder Störung des Tieres zu vermeiden, beispielsweise durch Sediment Resuspension. das Abtasten zu beginnen, durchsticht ein Taucher ein Septum in den Auffangbehälter mit einer Injektionsnadel auf die anhänge distale Ende jedes Rohrs, so dass der Außenwasserdruck das abgetastete Wasser in den Behälter durch das Entnahmerohr zu zwingen. das Ansaugen durch die Vakuum zuvor in den Fläschchen und durch die Druckdifferenz zwischen dem externen Wasser und dem evakuierten Probenbehälter geschaffen eingeleitet wird . Um eine saubere Sammlung von ausgeatmeten Wasser gewährleisten und ein versehentliches Absaugen von Ambi zu vermeidenent Wasser 16, muss die Wasser Abtastrate bei einer signifikant niedrigeren Rate (&lt;10%) als die excurrent Flussrate gehalten werden. Die Saugleistung wird durch die Länge des Rohres und dessen Innendurchmesser (ID) gesteuert. Der kleine Innendurchmesser sorgt auch für eine vernachlässigbare Totvolumen (&lt;200 &amp; mgr; l pro Meter Leitung). Probenahme über längere Zeiträume (Minuten bis Stunden) ermöglicht es, die inhärente Lückenhaftigkeit der meisten Substanzen von Interesse zu integrieren. Um sicherzustellen , dass die Proben werden in längeren Unterwasserprobenahme Sitzungen sowie für den Transport zum Labor ausreichend konserviert, eine in situ Filtration wird für empfindliche Analyte empfohlen. Die Auswahl der Stichprobenbehälter, Filtersystem und Schläuche werden durch die Studie Organismen und der spezifischen Fragestellung bestimmt. Das Protokoll weiter unten beschrieben wird davon ausgegangen , dass eine vollständige metabolische Profil von Interesse ist (für eine Übersicht siehe Abbildung 2). Jedoch erlaubt die modulare Natur des Protokolls foder einfache Änderung aufzunehmen einfacher oder sogar sehr unterschiedliche Stichprobenpläne. Für eine vollständige metabolische Profil sollte die Probenahmeprotokoll die folgenden Schritte umfassen: (1) Strömungsvisualisierung; (2) Die Probenahme ultra Plankton Fütterung (Plankton &lt;10 &amp; mgr; m); (3) Die Probenahme anorganische Nährstoffe Aufnahme und Ausscheidung (in-line Filter verwendet wird); (4) Die Probenahme gelösten organischen Aufnahme und Ausscheidung (unter Verwendung von in-line-Filter); (5) Particulate Fütterung und Ausscheidung (unter Verwendung von in-line-Filter); (6) Wiederholen Sie Schritt 2 (Ultra-Plankton Fütterung als Qualitätskontrolle); (7) Visualisierung fließen. Wenn logistisch machbar, wird empfohlen , dass die metabolische Profilmessungen mit Pumprate kombiniert werden (zB Verfahren der Farbstofffront Geschwindigkeit, in 16) sowie mit der Atmung Messungen. Diese Messungen werden am besten am Anfang und am Ende der Stichprobensitzung genommen. Für die Atmung Messung sind unter Wasser Optodenbasis oder Mikroelektroden bevorzugt.

<table><tbody><tr><td>GorillaPod, Original</td><td>Joby</td><td>GP000001</td><td>flexible portable tripod&amp;nbsp;</td></tr><tr><td>Flangeless Ferrule</td><td>IDEX Health &amp;amp; Science&amp;nbsp;</td><td>P-200X</td><td>1/16&quot; in Blue/pk</td></tr><tr><td>Male Nut</td><td>IDEX Health &amp;amp; Science</td><td>P-205X&amp;nbsp;</td><td>1/16&quot; in Green/10&amp;nbsp;pk</td></tr><tr><td>Female to Female Luer</td><td>IDEX Health &amp;amp; Science&amp;nbsp;</td><td>P-658</td><td></td></tr><tr><td>Female-Male Luer</td><td>IDEX Health &amp;amp; Science&amp;nbsp;</td><td>P-655</td><td></td></tr><tr><td>Peek Tubing (254&amp;nbsp;&amp;micro;m ID)</td><td>IDEX Health &amp;amp; Science&amp;nbsp;</td><td>1531</td><td>1/16&quot; OD x 0.01&amp;nbsp;in ID x 5&amp;nbsp;ft length. Alternative ID can be used.</td></tr><tr><td>Two component resin epoxy</td><td>IVEGOR</td><td>9257</td><td>Mix well the two component resin before use</td></tr><tr><td>(TOC) EPA Vials</td><td>Cole -Parmer</td><td>03756-20</td><td>40 ml glass vials. Manufactured also by Thomas Scientific (ref. number 9711F09).</td></tr><tr><td>HDPE Vials</td><td>Wheaton</td><td>986701 (E78620)</td><td>20 ml high-density polyethylene vials</td></tr><tr><td>Vacuette Z no additive</td><td>Greiner bio-one</td><td>455001</td><td>pre-vacuum by the manufacturer&amp;nbsp;</td></tr><tr><td>Septum Sample Bottles</td><td>Thomas Scientific</td><td>1755C01</td><td>250 ml glass bottles&amp;nbsp;</td></tr><tr><td>Septum Cap 1</td><td>Wheaton</td><td>W240844SP (E7865R)</td><td>22-400 for HDPE vials&amp;nbsp;</td></tr><tr><td>Septum Cap 2</td><td>Wheaton</td><td>W240846 (1078-5553)</td><td>24-400 for glass vials and bottles. Also manufactured by Thermo Scientific National (ref. 03-377-42).</td></tr><tr><td>In-line stainless steel Swinney Filter holders</td><td>Pall</td><td>516-9067</td><td>13 mm of diameter</td></tr><tr><td>PTFE Seal Washer</td><td>Pall&amp;nbsp;</td><td>516-8064</td><td>ring for stainless steel filter holders</td></tr><tr><td>TCLP Glass Filters</td><td>Pall&amp;nbsp;</td><td>516-9126</td><td>binder-free glass fiber filters, 13&amp;nbsp;mm of diameter, pore size 0.7&amp;nbsp;&amp;micro;m</td></tr><tr><td>Polycarbonate Filter Holders</td><td>Cole -Parmer</td><td>17295</td><td>13 mm of diameter</td></tr><tr><td>Isopore Membrane Filters</td><td>Millipore</td><td>GTTP01300</td><td>13&amp;nbsp;mm of diameter, pore size 0.2&amp;nbsp;&amp;micro;m</td></tr><tr><td>Contrad 70 Solution&amp;nbsp;</td><td>Decon Labs</td><td>1002</td><td>highly soluble basic mix of anionic and non-ionic surfactant solution&amp;nbsp;</td></tr><tr><td>Sterile Syringe Filters</td><td>VWR International Eurolab S.L.</td><td>514-0061P</td><td>25&amp;nbsp;mm of diameter , pore size 0.2&amp;nbsp;&amp;micro;m</td></tr><tr><td>Fluorescein</td><td>Sigma-Aldrich</td><td>(old ref.28802) 46955-100G</td><td>100&amp;nbsp;g</td></tr><tr><td>Holdex, disposable,sterile</td><td>Greiner bio-one</td><td>450263</td><td>sterile, single-use tube holder with off-center luer for Vacuette</td></tr><tr><td>Sterile Needles</td><td>IcoGammaPlus</td><td>5160</td><td>0.7&amp;nbsp;mm x 30&amp;nbsp;mm</td></tr><tr><td>Cryovials Nalgene</td><td>Nalgene</td><td>V5007(Cat. No.5000-0020)</td><td>2&amp;nbsp;ml</td></tr><tr><td>Cryobox carton&amp;nbsp;</td><td>Rubilabor</td><td>M-600</td><td>145x145x55&amp;nbsp;mm p/microtube 1.5&amp;nbsp;ml</td></tr><tr><td>Orthophosphoric Acid</td><td>Sigma</td><td>79617 or Alternatively use Ultra-Pure Hydrochloric acid, final concertation 0.1%</td><td></td></tr><tr><td>Paraformaldehyde</td><td>Sigma</td><td>P6148</td><td>500&amp;nbsp;g</td></tr><tr><td>Glutaraldehyde</td><td>Merck</td><td>8,206,031,000</td><td>25%, 1 L</td></tr><tr><td>Hand Vacuum Pump&amp;nbsp;</td><td>B&amp;uuml;rkle&amp;nbsp;</td><td>5620-2181 or Alternatively use standard laboratory vacuum pump</td><td></td></tr></tbody></table>

vacusip, an improved inex method for in situ measurement of particulate and dissolved compounds processed by active suspension feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.

Optimierung von Meerwasser Erhebungsmethoden Die Auswahl der Sammler Fläschchen und Reinigungsverfahren VACUSIP-kompatible Auffanggefäße sollte ein Septum, die Probenahme eingeleitet werden können, indem sie mit einer Spritzennadel Piercing. Sie sollten die erhöhten Unterwasserdruck (2-3 bar bei typischen Taucharbeitst...

Watch this Scientific Journal Video about VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders at JoVE.com

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

VACUSIP eine verbesserte Inex Methode für<em&gt; In Situ</em&gt; Messung von partikulären und gelösten Verbindungen, die durch Active Suspension Feeders Verarbeitet

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

vacusip-an-improved-inex-method-for-situ-measurement-particulate

Research

JoVE Journal

Environment

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

10.8K Aufrufe.  Centre d’Estudis Avançats de Blanes (CEAB-CSIC). We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

Serie: VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

Quantitatively Measuring In situ Flows using a Self-Contained Underwater Velocimetry Apparatus (SCUVA)

The ability to directly measure velocity fields in a fluid environment is necessary to provide empirical data for studies in fields as diverse as oceanography, ecology, biology, and fluid mechanics. Field measurements introduce practical challenges such as environmental conditions, animal availability, and the need for field-compatible measurement techniques. To avoid these challenges, scientists typically use controlled laboratory environments to study animal-fluid interactions. However, it is reasonable to question whether one can extrapolate natural behavior (i.e., that which occurs in the field) from laboratory measurements. Therefore, in situ quantitative flow measurements are needed to accurately describe animal swimming in their natural environment.
We designed a self-contained, portable device that operates independent of any connection to the surface, and can provide quantitative measurements of the flow field surrounding an animal. This apparatus, a self-contained underwater velocimetry apparatus (SCUVA), can be operated by a single scuba diver in depths up to 40 m. Due to the added complexity inherent of field conditions, additional considerations and preparation are required when compared to laboratory measurements. These considerations include, but are not limited to, operator motion, predicting position of swimming targets, available natural suspended particulate, and orientation of SCUVA relative to the flow of interest. The following protocol is intended to address these common field challenges and to maximize measurement success.

Quantitatively Measuring In situ Flows using a Self-Contained Underwater Velocimetry Apparatus SCUVA

This protocol provides instructions on how to use a self-contained underwater velocimetry apparatus (SCUVA), which is designed for quantification of in situ animal-generated flows. In addition, this protocol addresses challenges posed by field conditions, and includes operator motion, predicting position of animals, and orientation of SCUVA.

Quantitativen Messung In situ Flows mit einem Self-Contained Underwater Velocimetry Apparatus (SCUVA)

The ability to directly measure velocity fields in a fluid environment is necessary to provide empirical data for studies in fields as diverse as ...

Forschung

Biotechnik

Clean Sampling and Analysis of River and Estuarine Waters for Trace Metal Studies

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination control. Developments of some techniques aiming to reduce trace metal contamination in the last couple of decades have resulted in concentrations reported now being orders of magnitude lower than those in the past. These low concentrations often necessitate preconcentration of water samples prior to instrumental analysis of samples. Since contamination can appear in all phases of trace metal analyses, including sample collection (and during preparation of sampling containers), storage and handling, pretreatments, and instrumental analysis, specific care needs to be taken in order to reduce contamination levels at all steps. The effort to develop and utilize "clean techniques" in trace metal studies allows scientists to investigate trace metal distributions and chemical and biological behavior in greater details. This advancement also provides the required accuracy and precision of trace metal data allowing for environmental conditions to be related to trace metal concentrations in aquatic environments.
This protocol that is presented here details needed materials for sample preparation, sample collection, sample pretreatment including preconcentration, and instrumental analysis. By reducing contamination throughout all phases mentioned above for trace metal analysis, much lower detection limits and thus accuracy can be achieved. The effectiveness of "clean techniques" is further demonstrated using low field blanks and good recoveries for standard reference material. The data quality that can be obtained thus enables the assessment of trace metal distributions and their relationships to environmental parameters.

Special care using "clean techniques" is required to properly collect and process water samples for trace metal studies in aquatic environments. A protocol for sampling, processing, and analytical procedures with the aim of obtaining reliable environmental monitoring data and results with high sensitivity for detailed trace metal studies is presented.

Saubere Probenahme und Analyse von River und Estuarine Waters für Spurenmetallforschung

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination ...

Umwelt

Continuous Instream Monitoring of Nutrients and Sediment in Agricultural Watersheds

Pollutant concentrations and loads in watersheds vary considerably with time and space. Accurate and timely information on the magnitude of pollutants in water resources is a prerequisite for understanding the drivers of the pollutant loads and for making informed water resource management decisions. The commonly used &#34;grab sampling&#34; method provides the concentrations of pollutants at the time of sampling (i.e., a snapshot concentration) and may under- or overpredict the pollutant concentrations and loads. Continuous monitoring of nutrients and sediment has recently received more attention due to advances in computing, sensing technology, and storage devices. This protocol demonstrates the use of sensors, sondes, and instrumentation to continuously monitor in situ nitrate, ammonium, turbidity, pH, conductivity, temperature, and dissolved oxygen (DO) and to calculate the loads from two streams (ditches) in two agricultural watersheds. With the proper calibration, maintenance, and operation of sensors and sondes, good water quality data can be obtained by overcoming challenging conditions such as fouling and debris buildup. The method can also be used in watersheds of various sizes and characterized by agricultural, forested, and/or urban land.

With the advancement of technology and the rise in end-user expectations, the need and use of higher temporal resolution data for pollutant load estimation has increased. This protocol describes a method for continuous in situ water quality monitoring to obtain higher temporal resolution data for informed water resource management decisions.

Kontinuierliche Instream Überwachung von Nährstoffen und Sediment in landwirtschaftlichen Wasserscheiden

Pollutant concentrations and loads in watersheds vary considerably with time and space. Accurate and timely information on the magnitude of pollutants in ...

Capturing Flow-weighted Water and Suspended Particulates from Agricultural Canals During Drainage Events

The purpose of this study is to describe the methods used to capture flow-weighted water and suspended particulates from farm canals during drainage discharge events. Farm canals can be enriched by nutrients such as phosphorus (P) that are susceptible to transport. Phosphorus in the form of suspended particulates can significantly contribute to the overall P loads in drainage water. A settling tank experiment was conducted to capture suspended particulates during discrete drainage events. Farm canal discharge water was collected in a series of two 200 L settling tanks over the entire duration of the drainage event, so as to represent a composite subsample of the water being discharged. Imhoff settling cones are ultimately used to settle out the suspended particulates. This is achieved by siphoning water from the settling tanks via the cones. The particulates are then collected for physico-chemical analyses.

Nutrients present in particulate form can contribute significantly to the overall loads in agricultural drainage waters. This study describes a novel method to capture flow-weighted water and suspended particulates from farm canal drainage over the entire duration of the drainage event.

Erfassung von Wasser fließen gewichtet und Schwebestaub aus landwirtschaftlichen Kanäle bei Drainage-Veranstaltungen

The purpose of this study is to describe the methods used to capture flow-weighted water and suspended particulates from farm canals during drainage ...

Continuous Hydrologic and Water Quality Monitoring of Vernal Ponds

Vernal ponds, also referred to as vernal pools, provide critical ecosystem services and habitat for a variety of threatened and endangered species. However, they are vulnerable parts of the landscapes that are often poorly understood and understudied. Land use and management practices, as well as climate change are thought to be a contribution to the global amphibian decline. However, more research is needed to understand the extent of these impacts. Here, we present methodology for characterizing a vernal pond's morphology and detail a monitoring station that can be used to collect water quantity and quality data over the duration of a vernal pond's hydroperiod. We provide methodology for how to conduct field surveys to characterize the morphology and develop stage-storage curves for a vernal pond. Additionally, we provide methodology for monitoring the water level, temperature, pH, oxidation-reduction potential, dissolved oxygen, and electrical conductivity of water in a vernal pond, as well as monitoring rainfall data. This information can be used to better quantify the ecosystem services that vernal ponds provide and the impacts of anthropogenic activities on their ability to provide these services.

Understanding the ecosystem services and processes provided by vernal ponds and the impacts of anthropogenic activities on their ability to provide these services requires intensive hydrologic monitoring. This sampling protocol using in-situ monitoring equipment was developed to understand the impact of anthropogenic activities on water levels and quality.

Kontinuierliche hydrologischen und Qualitätsüberwachung von Vernal Teichen

Vernal ponds, also referred to as vernal pools, provide critical ecosystem services and habitat for a variety of threatened and endangered species. ...

An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles

The distributions of many trace elements in the ocean are strongly associated with the growth, death, and re-mineralization of marine plankton and those of suspended/sinking particles. Here, we present an all plastic (Polypropylene and Polycarbonate), multi-layer filtration system for collection of suspended particulate matter (SPM) at sea. This ultra-clean sampling device has been designed and developed specifically for trace element studies. Meticulous selection of all non-metallic materials and utilization of an in-line flow-through procedure minimizes any possible metal contamination during sampling. This system has been successfully tested and tweaked for determining trace metals (e.g., Fe, Al, Mn, Cd, Cu, Ni) on particles of varying size in coastal and open ocean waters. Results from the South China Sea at the South East Asia Time-Series (SEATS) station indicate that diurnal variations and spatial distribution of plankton in the euphotic zone can be easily resolved and recognized. Chemical analysis of size-fractionated particles in surface waters of the Taiwan Strait suggests that the larger particles (&#62;153 &#181;m) were mostly biologically derived, while the smaller particles (10 - 63 &#181;m) were mostly composed of inorganic matter. Apart from Cd, the concentrations of metals (Fe, Al, Mn, Cu, Ni) decreased with increasing size.

Plankton and suspended particles play a major role in the biogeochemical cycles in the ocean. Here, we provide an ultra-clean, low stress method for the collection of various sizes of particles and plankton at sea with the capability of handling large volumes of seawater.

Eine Ultra-clean Multilayer Vorrichtung zum Sammeln von Größe fraktioniert Meeresplankton und Schwebeteilchen

The distributions of many trace elements in the ocean are strongly associated with the growth, death, and re-mineralization of marine plankton and those ...

Design and Use of an Apparatus for Quantifying Bivalve Suspension Feeding at Sea

As shellfish aquaculture moves from coastal embayments and estuaries to offshore locations, the need to quantify ecosystem interactions of farmed bivalves (i.e., mussels, oysters, and clams) presents new challenges. Quantitative data on the feeding behavior of suspension-feeding mollusks is necessary to determine important ecosystem interactions of offshore shellfish farms, including their carrying capacity, the competition with the zooplankton community, the availability of trophic resources at different depths, and the deposition to the benthos. The biodeposition method is used to quantify feeding variables in suspension-feeding bivalves in a natural setting and represents a more realistic proxy than laboratory experiments. This method, however, relies upon a stable platform to satisfy the requirements that water flow rates supplied to the shellfish remain constant and the bivalves are undisturbed. A flow-through device and process for using the biodeposition method to quantify the feeding of bivalve mollusks were modified from a land-based format for shipboard use by building a two-dimensional gimbal table around the device. Planimeter data reveal a minimal pitch and yaw of the chambers containing the test shellfish despite boat motion, the flow rates within the chambers remain constant, and operators are able to collect the biodeposits (feces and pseudofeces) with sufficient consistency to obtain accurate measurements of the bivalve clearance, filtration, selection, ingestion, rejection, and absorption at offshore shellfish aquaculture sites.

A flow-through device for using the biodeposition method to quantify filtration and feeding behavior of bivalve mollusks was modified for shipboard use. A two-dimensional gimbal table built around the device isolates the apparatus from boat motion, thereby allowing the accurate quantification of bivalve filtration variables at offshore shellfish aquaculture sites.

Gestaltung und Nutzung eines Apparates zur Quantifizierung der zweischaligen Suspension Fütterung auf hoher See

As shellfish aquaculture moves from coastal embayments and estuaries to offshore locations, the need to quantify ecosystem interactions of farmed bivalves ...

In Situ Chemotaxis Assay to Examine Microbial Behavior in Aquatic Ecosystems

Microbial behaviors, such as motility and chemotaxis (the ability of a cell to alter its movement in response to a chemical gradient), are widespread across the bacterial and archaeal domains. Chemotaxis can result in substantial resource acquisition advantages in heterogeneous environments. It also plays a crucial role in symbiotic interactions, disease, and global processes, such as biogeochemical cycling. However, current techniques restrict chemotaxis research to the laboratory and are not easily applicable in the field. Presented here is a step-by-step protocol for the deployment of the in situ chemotaxis assay (ISCA), a device that enables robust interrogation of microbial chemotaxis directly in the natural environment. The ISCA is a microfluidic device consisting of a 20 well array, in which chemicals of interest can be loaded. Once deployed in aqueous environments, chemicals diffuse out of the wells, creating concentration gradients that microbes sense and respond to by swimming into the wells via chemotaxis. The well contents can then be sampled and used to (1) quantify strength of the chemotactic responses to specific compounds through flow cytometry, (2) isolate and culture responsive microorganisms, and (3) characterize the identity and genomic potential of the responding populations through molecular techniques. The ISCA is a flexible platform that can be deployed in any system with an aqueous phase, including marine, freshwater, and soil environments.

Presented here is the protocol for an in situ chemotaxis assay, a recently developed microfluidic device that enables studies of microbial behavior directly in the environment.

In Situ Chemotaxis Assay zur Untersuchung des mikrobiellen Verhaltens in aquatischen Ökosystemen

Microbial behaviors, such as motility and chemotaxis (the ability of a cell to alter its movement in response to a chemical gradient), are widespread ...

Microparticle Manipulation by Standing Surface Acoustic Waves with Dual-frequency Excitations

We demonstrate a method for increasing the tuning ability of a standing surface acoustic wave (SSAW) for microparticles manipulation in a lab-on-a-chip (LOC) system. The simultaneous excitation of the fundamental frequency and its third harmonic, which is termed as dual-frequency excitation, to a pair of interdigital transducers (IDTs) could generate a new type of standing acoustic waves in a microfluidic channel. Varying the power and the phase in the dual-frequency excitation signals results in a reconfigurable field of the acoustic radiation force applied to the microparticles across the microchannel (e.g., the number and location of the pressure nodes and the microparticle concentrations at the corresponding pressure nodes). This article demonstrates that the motion time of the microparticle to only one pressure node can be reduced ~2-fold at the power ratio of the fundamental frequency greater than ~90%. In contrast, there are three pressure nodes in the microchannel if less than this threshold. Furthermore, adjusting the initial phase between the fundamental frequency and the third harmonic results in different motion rates of the three SSAW pressure nodes, as well as in the percentage of microparticles at each pressure node in the microchannel. There is a good agreement between the experimental observation and the numerical predictions. This novel excitation method can easily and non-invasively integrate into the LOC system, with a wide tenability and only a few changes to the experimental set-up.

A protocol for manipulating the microparticles in a microfluidic channel with a dual-frequency excitation is presented.

Mikropartikel Manipulation durch ständigen akustischen Oberflächenwellen mit Zweifrequenz-Erregungen

We demonstrate a method for increasing the tuning ability of a standing surface acoustic wave (SSAW) for microparticles manipulation in a lab-on-a-chip ...

Ingenieurwesen

Dissolved Solute Sampling Across an Oxic-Anoxic Soil-Water Interface Using Microdialysis Profilers

Biogeochemical processes shift rapidly in both spatial (millimeter scale) and temporal (hour scale to day scale) dimensions at the oxic-anoxic interface in response to disturbances. Deciphering the rapid biogeochemical changes requires in situ, minimally invasive tools with high spatial and temporal sampling resolution. However, the available passive sampling devices are not very useful in many cases either due to their disposable nature or the complexity and extensive workload for sample preparation.
To address this problem, a microdialysis profiler with 33 individual polyethersulfone nanomembrane tubes (semipermeable, &#60;20 nm pore size) integrated into the one-dimensional skeleton (60 mm) was established to iteratively sample the dissolved compounds in porewater across the soil-water interface at a high resolution of 1.8 mm (outer diameter plus one spacing, i.e., 0.1 mm between probes). The sampling mechanism is based on the principle of concentration gradient diffusion. The automatic loading of degassed water allows minimal disturbance to the chemical species across the oxic-anoxic interface.
This paper describes the procedures of device setup and continuous porewater sampling across the soil-water interface on a daily basis. Concentration-depth profiles were selectively measured before (on Day 6) and after (on Day 7) disturbances induced by irrigation. The results showed that concentration-depth profiles were undergoing rapid changes, especially for redox-sensitive elements (i.e., iron and arsenic). These protocols can help investigate the biogeochemical responses across the soil-water interface under various disturbances caused by physical, chemical, and biological factors. The paper thoroughly discusses the advantages and disadvantages of this method for potential use in the environmental sciences.

A microdialysis profiler is described to sample dissolved porewater solutes across an oxic-anoxic soil-water interface in situ with minimal disturbance. This device is designed to capture rapid changes in concentration-depth profiles induced by disturbances at the soil-water interface and beyond.

Probenahme gelöster gelöster Stoffe an einer oxisch-anoxischen Boden-Wasser-Grenzfläche mit Mikrodialyse-Profilern

Biogeochemical processes shift rapidly in both spatial (millimeter scale) and temporal (hour scale to day scale) dimensions at the oxic-anoxic interface ...

VACUSIP eine verbesserte Inex Methode für

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