Name: a fish-feeding laboratory bioassay to assess the antipredatory activity of secondary metabolites from the tissues of marine organisms
Uploaded: 2015-01-11T14:00:00
Description: Watch this Scientific Journal Video about A Fish-feeding Laboratory Bioassay to Assess the Antipredatory Activity of Secondary Metabolites from the Tissues of Marine Organisms at JoVE.com

We thank James Maeda and Aaron Cooke for assistance with the filming and editing of this video. Funding was provided by the National Science Foundation (OCE-0550468, 1029515).

HINWEIS: Schritt 3 dieses Protokolls betrifft Wirbeltier Themen. Das Verfahren ist so konzipiert, dass die Tiere erhalten die humane Behandlung möglich und wurde von der Institutional Animal Care und Verwenden Committee (IACUC) an der University of North Carolina Wilmington zugelassen. 1) Gewebeentnahme <ol><li> Verwenden Gewebe, das in seinem natürlichen Zustand der Flüssigkeitszufuhr und nicht gequetscht, ausgetrocknete oder übermäßig nass ist, da dies die Volumenkonzentration von ...

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Das hier beschriebene Verfahren stellt ein relativ einfaches, ökologisch relevanten Laborprotokoll für die Bewertung antipredatory chemische Abwehr in Meeresorganismen. Hier beschreiben wir die wichtigsten Kriterien, die von diesem Satz von Methoden erfüllt werden: (1) Geeignete Raubtier. Das Fütterungstest verwendet den bluehead Lippfische, Thalassoma bifasciatum, einer der am häufigsten vorkommenden Fische auf Korallenriffe in der Karibik. Die bluehead ist ein Genera...

Chemische Ökologie durch die Zusammenarbeit von Chemikern und Ökologen entwickelt. Während der Teildisziplin der terrestrischen chemischen Ökologie gibt es schon seit einiger Zeit, dass der Meeres chemische Ökologie ist nur ein paar Jahrzehnte alt, hat aber wichtige Einblicke in die Evolutionsökologie und Gemeinschaftsstruktur von Meeresorganismen 8.1 zur Verfügung gestellt. Sie profitieren von den neuen Technologien von Tauchen und NMR-Spektroskopie, Organiker schnell erzeugt eine große Anzahl von Veröffentlichungen, die neuartige Stoffwechselprodukte von benthischen wirbellosen Meerestieren und Algen in den 1970er und 1980er Jahren 9. Unter der Annahme, dass sekundäre Pflanzenstoffe muss einen Zweck haben viele dieser Publikationen ist ohne empirische Beweise zugeschrieben ökologisch wichtigen Eigenschaften, neue Verbindungen zu dienen. Etwa zur gleichen Zeit wurden Ökologen auch die Vorteile der Einführung von Tauchen und Beschreibung der Verteilungen und Häufigkeiten von Bodentieren und Pflanzen früher her bekanntbin relativ unwirksam Probenahmeverfahren wie Baggerarbeiten. Die Annahme dieser Forscher war, dass nichts festsitzenden und weichen Körper muss chemisch verteidigt werden Verbrauch von Raubtieren 10 zu vermeiden. In dem Bemühen, den Empirismus zu dem, was sonst beschreibende Arbeiten an Arten Häufigkeiten einzuführen, begannen einige Ökologen Extrapolation chemische Abwehr von Toxizitätstests 11. Die meisten Toxizitätstests ging es um die Exposition der ganze Fische oder andere Organismen, um wässrige Suspensionen von rohen organischen Extrakte der wirbellosen Gewebe, mit anschließender Bestimmung der Trockenmasse Konzentrationen von Extrakten für die Tötung der Hälfte der Testorganismen verantwortlich. Allerdings sind Toxizitätstests nicht emulieren die Art, in der potenzielle Räuber wahr Beute unter natürlichen Bedingungen, und nachfolgende Studien haben keinen Zusammenhang zwischen Toxizität und Schmackhaftigkeit 12-13 gefunden. Es überrascht, dass Publikationen in renommierten Fachzeitschriften verwendet Techniken, die wenig oder gar keine ecological Relevanz 14-15 und dass diese Studien sind immer noch weit zitiert heute. Es ist noch alarmierender zu beachten, dass Studien, die auf Toxizitätsdaten weiterhin veröffentlicht 16-18 werden. Die hier beschriebene Bioassay-Methode wurde in den späten 1980er Jahren entwickelt, um eine ökologisch relevanten Ansatz für die Meeres chemischen Ökologen sorgen zu antipredatory chemische Abwehr zu bewerten. Das Verfahren erfordert ein Modell Raubtier, um eine rohe organische Extrakt aus den Zielorganismus zu einem natürlichen Konzentration in einer ernährungs vergleichbaren Lebensmittelmatrix probieren und bietet Schmackhaftigkeit Daten, die ökologisch sinnvoller als Toxizitätsdaten. Der allgemeine Ansatz zur Bewertung der antipredatory Aktivität der Gewebe von Meeresorganismen enthält vier wichtige Kriterien: (1) eine angemessene Generalist Räuber müssen in Fütterungstests verwendet werden, (2) Bio-Stoffwechselprodukte aller Polaritäten müssen erschöpfend aus dem Gewebe der extrahiert werden Zielorganismus, (3) die Metaboliten muss be an der gleichen Volumenkonzentration in einer ernährungs geeigneten experimentellen Nahrung vermischt als in dem Organismus, aus dem sie entnommen wurden, und (4) das experimentelle Design und statistische Ansatz müssen einen sinn Metrik relativ distastefulness hindeuten. Die unten aufgeführten Verfahren ist speziell auf antipredatory chemische Abwehr in der Karibik wirbellose Meerestiere zu bewerten. Wir beschäftigen die bluehead Lippfische, Thalassoma bifasciatum, als Modell Raubfische, weil diese Art häufig auf karibischen Korallenriffe und ist dafür bekannt, ein breites Sortiment von Makrozoobenthos 19 kosten. Gewebe aus dem Zielorganismus wird zuerst extrahiert, dann kombiniert mit einer Futtermischung, und schließlich, um Gruppen von T. angeboten bifasciatum zu beobachten, ob sie den Extrakt behandelten Lebensmitteln abzulehnen. Assay-Daten mit dieser Methode haben wichtige Einblicke in die Defensive Chemie mariner Organismen 12,20-21, l bereitgestelltife Geschichte Kompromisse 22-24 und Gemeinschaftsökologie 25-26.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Dichloromethane</td>
			<td>Fisher Scientific</td>
			<td>D37-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Fisher Scientific</td>
			<td>A41220</td>
			<td></td>
		</tr>
		<tr>
			<td>Anhydrous Calcium Chloride</td>
			<td>Fisher Scientific</td>
			<td>C614-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Cryocool Heat Transfer Fluid</td>
			<td>Fisher Scientific</td>
			<td>20-548-146</td>
			<td>For vacuum concentrator</td>
		</tr>
		<tr>
			<td>Alginic Acid Sodium Salt High Viscosity</td>
			<td>MP Biomedicals</td>
			<td>154723</td>
			<td></td>
		</tr>
		<tr>
			<td>Squid mantle rings</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Can be purchased at grocery store</td>
		</tr>
		<tr>
			<td>Denatonium benzoate</td>
			<td>Aldrich</td>
			<td>D5765</td>
			<td></td>
		</tr>
		<tr>
			<td>50 ml graduated centrifuge tube</td>
			<td>Fisher Scientific</td>
			<td>14-432-22</td>
			<td></td>
		</tr>
		<tr>
			<td>20 ml scintillation vial</td>
			<td>Fisher Scientific</td>
			<td>03-337-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Disposable Pasteur pipets</td>
			<td>Fisher Scientific</td>
			<td>13-678-20D</td>
			<td></td>
		</tr>
		<tr>
			<td>Rubber bulbs for Pasteur pipets</td>
			<td>Fisher Scientific</td>
			<td>03-448-24</td>
			<td></td>
		</tr>
		<tr>
			<td>Red bulbs for pellet delivery</td>
			<td>Fisher Scientific</td>
			<td>03-448-27</td>
			<td></td>
		</tr>
		<tr>
			<td>250 ml round-bottom flask</td>
			<td>Fisher Scientific</td>
			<td>10-067E</td>
			<td></td>
		</tr>
		<tr>
			<td>Scintillation vial adapter for rotavap</td>
			<td>Fisher Scientific</td>
			<td>K747130-1324</td>
			<td></td>
		</tr>
		<tr>
			<td>Weightboats</td>
			<td>Fisher Scientific</td>
			<td>02-202B</td>
			<td></td>
		</tr>
		<tr>
			<td>Microspatula</td>
			<td>Fisher Scientific</td>
			<td>21-401-10</td>
			<td></td>
		</tr>
		<tr>
			<td>5 ml graduated syringe</td>
			<td>Fisher Scientific</td>
			<td>14-817-53</td>
			<td></td>
		</tr>
		<tr>
			<td>10 ml graduated syringe</td>
			<td>Fisher Scientific</td>
			<td>14-817-54</td>
			<td></td>
		</tr>
		<tr>
			<td>Razor blade</td>
			<td>Fisher Scientific</td>
			<td>S17302</td>
			<td></td>
		</tr>
	</tbody>
</table>

a fish-feeding laboratory bioassay to assess the antipredatory activity of secondary metabolites from the tissues of marine organisms

Marine chemical ecology is a young discipline, having emerged from the collaboration of natural products chemists and marine ecologists in the 1980s with the goal of examining the ecological functions of secondary metabolites from the tissues of marine organisms. The result has been a progression of protocols that have increasingly refined the ecological relevance of the experimental approach. Here we present the most up-to-date version of a fish-feeding laboratory bioassay that enables investigators to assess the antipredatory activity of secondary metabolites from the tissues of marine organisms. Organic metabolites of all polarities are exhaustively extracted from the tissue of the target organism and reconstituted at natural concentrations in a nutritionally appropriate food matrix. Experimental food pellets are presented to a generalist predator in laboratory feeding assays to assess the antipredatory activity of the extract. The procedure described herein uses the bluehead, Thalassoma bifasciatum, to test the palatability of Caribbean marine invertebrates; however, the design may be readily adapted to other systems. Results obtained using this laboratory assay are an important prelude to field experiments that rely on the feeding responses of a full complement of potential predators. Additionally, this bioassay can be used to direct the isolation of feeding-deterrent metabolites through bioassay-guided fractionation. This feeding bioassay has advanced our understanding of the factors that control the distribution and abundance of marine invertebrates on Caribbean coral reefs and may inform investigations in diverse fields of inquiry, including pharmacology, biotechnology, and evolutionary ecology.

Hier berichten wir über Ergebnisse der biologischen Tests für sechs Arten von gemeinsamen karibischen Schwämmen (Abbildung 2). Diese Daten wurden ursprünglich im Jahr 1995 von Pawlik et al. 12 veröffentlicht und zeigen, die Macht dieser Ansatz für die Unterschiede in der chemischen Abwehrstrategien unter Co-auftretende Taxa Umfrage. Die Ergebnisse wurden als mittlere Anzahl von Futterpellets Standardfehler (SE) für die einzelnen Arten gegessen + wiesen. Fast keine Pellets wurd...

Watch this Scientific Journal Video about A Fish-feeding Laboratory Bioassay to Assess the Antipredatory Activity of Secondary Metabolites from the Tissues of Marine Organisms at JoVE.com

A Fish-feeding Laboratory Bioassay to Assess the Antipredatory Activity of Secondary Metabolites from the Tissues of Marine Organisms

Dieser Bioassay verwendet ein Modell Raubfische, die Anwesenheit von Futter Abschreckung Metaboliten aus organischen Extrakte der Gewebe von Meeresorganismen auf natürlichen Konzentrationen mit einem ernährungsphysiologisch vergleichbaren Lebensmittelmatrix zu bewerten.

Ein Fisch-Fütterung Labor Bioassay die antipredatory Aktivität von Sekundärmetaboliten aus dem Gewebe mariner Organismen beurteilen

Marine chemical ecology is a young discipline, having emerged from the collaboration of natural products chemists and marine ecologists in the 1980s with ...

Research

JoVE Journal

Environment

Unraveling the Unseen Players in the Ocean - A Field Guide to Water Chemistry and Marine Microbiology

Entwirren der unsichtbaren Spieler im Ozean - ein Feld Führer zu Wasserchemie und Marine Mikrobiologie

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Forschung

Umwelt

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Laboratory Simulation of an Iron(II)-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria

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Labor Simulation eines Eisen (II) -reichen Präkambrium Meeres Upwelling-System das Wachstum von Bakterien zu erkunden Photosynthetic

A conventional concept for the deposition of some Precambrian Banded Iron Formations (BIF) proceeds on the assumption that ferrous iron [Fe(II)] upwelling ...

Ecotoxicological Method with Marine Bacteria Vibrio anguillarum to Evaluate the Acute Toxicity of Environmental Contaminants

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Bacteria are an important component of the ecosystem, and microbial community alterations can have a significant effect on biogeochemical cycling and food ...

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Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment ...

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The use of video camera systems in ecological studies of fish continues to gain traction as a viable, non-extractive method of measuring fish lengths and ...

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

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