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).

注：本协议的第3步涉及脊椎动物科目。该程序已被设计成动物获得了最人性化的治疗可能和已经批准的机构动物护理和使用委员会（IACUC）在北卡罗莱纳州威尔明顿的大学。 1）组织提取<ol><li>使用组织是在其天然水，而不是挤压，干涸的或过于潮湿的状态，因为这将改变次生代谢产物的体积浓度。切剁的组织，以片或片可插入到50ml离心管中。注意：新鲜组织可以在某些情?...

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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+of+a+resource+trade-off+between+growth+and+chemical+defenses+among+Caribbean+coral+reef+sponges.">Evidence of a resource trade-off between growth and chemical defenses among Caribbean coral reef sponges.</a> Marine Ecology Progress Series. 406, 71-78 (2010).</li><li>Leong, W., Pawlik, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+reproductive+patterns+among+7+Caribbean+sponge+species+does+not+reveal+a+resource+trade-off+with+chemical+defenses.">Comparison of reproductive patterns among 7 Caribbean sponge species does not reveal a resource trade-off with chemical defenses.</a> Journal of Experimental Marine Biology and Ecology. 401 (1-2), 80-84 (2011).</li><li>Pawlik, J. R., Loh, T. -. L., McMurray, S. E., Finelli, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sponge+Communities+on+Caribbean+Coral+Reefs+Are+Structured+by+Factors+That+Are+Top-Down,+Not+Bottom-Up.">Sponge Communities on Caribbean Coral Reefs Are Structured by Factors That Are Top-Down, Not Bottom-Up.</a> PLoS ONE. 8 (5), e62573 (2013).</li><li>Loh, T. -. L., Pawlik, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+defenses+and+resource+trade-offs+structure+sponge+communities+on+Caribbean+coral+reefs.">Chemical defenses and resource trade-offs structure sponge communities on Caribbean coral reefs.</a> Proceedings of the National Academy of Science. 111, 4151-4156 (2014).</li><li>Miller, A. M., Pawlik, J. 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R., Ilan, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+anti-predatory+defenses+of+Red+Sea+and+Caribbean+sponges.+I.+Chemical+defense.">Comparison of anti-predatory defenses of Red Sea and Caribbean sponges. I. Chemical defense.</a> Marine Ecology Progress Series. 252, 105-114 (2003).</li><li>Jones, A. C., Blum, J. E., Pawlik, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testing+for+defensive+synergy+in+Caribbean+sponges:+Bad+taste+or+glass+spicules.">Testing for defensive synergy in Caribbean sponges: Bad taste or glass spicules.</a> Journal of Experimental Marine Biology and Ecology. 322 (1), 67 (2005).</li></ol>

这里所描述的程序提供了一个相对简单的，生态的相关实验室的协议，以评估海洋生物antipredatory化学防御。在这里，我们回顾了由这套方法满足重要的标准： （1）适当的捕食者。这种喂养法采用双带锦鱼，Thalassoma bifasciatum，珊瑚礁整个加勒比地区最丰富的鱼类之一。该bluehead是一个通才食肉动物称为采样底栖无脊椎动物19各式各样。通才天敌是最好的?...

通过化学家和生态学家的合作化学生态学的发展。虽然陆地化学生态学的分支学科已经有一段时间了，那海洋化学生态学的只有几十年的历史，但已经提供了重要的见解海洋生物1-8的进化生态学和群落结构。以水肺潜水和核磁共振光谱的新兴技术的优势，有机化学迅速产生，描述上世纪70年代和80年代，从9底栖海洋无脊椎动物和藻类代谢产物的小说出版了大量。假设次级代谢产物必须为某种目的，许多这些出版物没有经验证据冲高重要生态价值属性的新化合物。大约在同一时间，生态学家还采取水肺潜水的来临优势和描述的分布和底栖动物和植物的前身来回的丰度米相对无效的抽样方法，如疏浚。这些研究人员的假设是，任何无柄和软体必须捍卫化学，以避免掠食者10消费。在努力引进经验什么是关于物种丰度，否则描述工作中，一些生态学家开始从毒性试验11推断的化学防御。大多数毒性测定法涉及的整鱼或其他生物体的脊椎动物组织的粗有机萃取水性悬浮液的曝光，随后确定负责杀死半数试验生物提取物的干重的浓度。然而，毒性试验不效仿其潜在的掠食者自然条件下感知猎物的方式，和随后的研究发现毒性和12-13适口性之间没有任何关系。令人惊讶的是，在著名期刊出版物使用具有很少或没有。生态技术升相关14-15，今天，这些研究仍然被广泛引用。而更令人震惊的要注意的是基于毒性数据研究继续出版16-18。这里所描述的生物测定方法的开发，在20世纪80年代后期，为海洋化工生态学家的生态相​​关的方法来评估antipredatory化学防御。该方法需要一个模型捕食样品从目标生物体的粗有机提取物在营养食品可比矩阵天然浓度，从而提供适口的数据，比毒性数据更加生态有意义。 一般的方法来评估海洋生物的组织的antipredatory活动包括四个重要的标准：（1）一个适当的通才捕食必须馈送测定法中使用，（2）所有极性有机代谢物必须详尽地从所述的组织中提取靶生物，（3）的代谢物必须BE在相同体积浓度混入营养合适的实验食品如在先前从其中提取，并且（4）的实验设计和统计方法必须提供一个有意义的度量，以指示相对于令人厌恶的生物体中发现。 下面的程序是专为评估加勒比海海洋无脊椎动物antipredatory化学防御。我们采用了双带锦鱼，Thalassoma bifasciatum，作为一种模式掠食性鱼类，因为这是种常见的对加勒比海的珊瑚礁，被称为采样底栖无脊椎动物19各式各样。从目标生物体组织被首先提取出来，然后与一种食物混合物，最后提供给T的组bifasciatum，观察他们是否拒绝提取物治疗食品。使用这种方法测定的数据提供了重要的见解海洋生物12,20-21，L的化学防御IFE历史权衡22-24和群落生态学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.

在这里，我们报告该生物测定6种常见加勒比海绵（ 图2）的结果。这些数据最初发表于1995年由Pawlik 等 12并证明了这种方法的力量调查共发生类群之间的化学防御策略的差异。结果被报告为标准误差（SE），针对每个物种食用+食物颗粒的平均数量。几乎没有颗粒被吃掉的测定与象耳海绵，Amphimedon牛鞭 ，并Aplysina cauliformis粗制有机提取物。与此相反，从Ca...

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

这种生物测定采用模型肉食性鱼类，以评估喂食威慑代谢产物从海洋生物的天然浓度的组织有机提取物用营养可比食品基质的存在。

了鱼喂食实验室生物测定，从海洋生物的组织评估次生代谢产物的Antipredatory活动

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

揭开看不见的玩家在海洋 - 一个领域指南水化学和海洋微生物

Here we introduce a series of thoroughly tested and well standardized research protocols adapted for use in remote marine environments. The sampling ...

科研

环境科学

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated (WATER-PAM) Fluorometry

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated WATER-PAM Fluorometry

小体积的生物测定评估细菌/浮游植物共培养使用水脉冲幅度调制（WATER-PAM）荧光

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be ...

Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples

自动凝胶尺寸选择，以提高下一代测序文库的质量，从环境水样准备

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA ...

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

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

VacuSIP，对改进的方法INEX<em&gt;原位</em&gt;颗粒物与测量溶解的化合物加工后主动悬挂给料机

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

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

Laboratory Simulation of an IronII-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria

一个铁（II）的实验室模拟的前寒武纪富海洋上升流系统探索光合细菌生长

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

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

生物毒理学方法与海洋细菌<em&gt;鳗弧菌</em&gt;评估环境污染物的急性毒性

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

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

从塑料颗粒中提取有机氯农药和塑料类型分析

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

Development of New Methods for Quantifying Fish Density Using Underwater Stereo-video Tools

利用水下立体视频工具量化鱼类密度的新方法研究进展

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 ...

The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

藤壶Balanus improvisus作为海洋模型的培养和基因表达

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

Long-term Video Tracking of Cohoused Aquatic Animals: A Case Study of the Daily Locomotor Activity of the Norway Lobster (Nephrops norvegicus)

Long-term Video Tracking of Cohoused Aquatic Animals: A Case Study of the Daily Locomotor Activity of the Norway Lobster Nephrops norvegicus

生态动物的长期视频跟踪: 挪威龙虾 (诺菲氏肾病) 日生物活动的案例研究

We present a protocol related to a video-tracking technique based on the background subtraction and image thresholding that makes it possible to ...