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).

참고 :이 프로토콜의 3 단계는 척추 동물의 과목을 포함한다. 절차는 동물이 가능한 가장 인도적인 치료를받을 수 있도록 설계되었으며, 노스 캐롤라이나 윌 밍턴 대학의 기관 동물 관리 및 사용위원회 (IACUC)에 의해 승인되었다. 1) 조직 추출 <ol><li> 이 이차 대사 산물의 체적 농도를 변경하므로 수화가 아닌 압착, 건조 아웃 또는 지나치게 젖은의 자연 상태에있는 조직을 사?...

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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>

본원에 기술 된 절차에 antipredatory 해양 생물 화학적 방어를 평가하기위한 상대적으로 간단하고, 생태 학적으로 중요한 실험 프로토콜을 제공한다. 여기에서 우리는 방법이 세트에 만족하는 중요한 기준을 검토 : (1) 적절한 육식 동물.이 공급 분석은 bluehead 놀래기과의 바닷물 고기, Thalassoma의 bifasciatum, 카리브해 전역의 산호초에 가장 풍부한 물고기 중 하나를 ?...

화학 생태학 화학자 및 생태 학자의 협력을 통해 개발했다. 지상파 화학 생태학의 subdiscipline 몇 시간 동안 주변되었습니다 있지만, 해양 화학 생태학의는 몇 십 년 오래지만 해양 생물 1-8의 진화 생태학과 사회 구조에 중요한 통찰력을 제공하고 있습니다. 스쿠버 다이빙과 NMR 분광법의 응급 기술을 활용, 유기 화학자들은 빠르게 1970 년대와 1980 년대 9 저서 무척추 동물과 조류의 소설 대사를 설명하는 출판물의 큰 숫자를 생성합니다. 이차 대사 산물 경험적 증거도없이 새로운 화합물에 생태 학적으로 중요한 특성을 관찰 작용이 책의 많은 어떤 목적을 봉사해야한다는 것을 가정. 약 동시에, 생태도 분포 이전에 이리저리 알려진 저서 동물과 식물의 풍부함을 스쿠버 다이빙의 출현을 활용하고 설명했다준설 상대적으로 비효율적 인 샘플링 방법을 해요. 이러한 연구자의 ​​가정은 고착 아무것도 부드러운 바디 화학적 육식 동물 (10)에 의해 소비를 방지하기 위해 방어해야이었다. 종 중 농도에 달리 설명 작업 사용했던 경험주의를 소개하는 노력의 일환으로, 일부 생태 독성 분석 (11)에서 화학 방어를 외삽하기 시작했다. 대부분의 독성 분석은 절반 분석 유기체를 죽이는에 대한 책임 추출물의 건조 질량 농도의 후속 결정으로, 전체 물고기 나 무척추 동물 조직의 원유 유기 추출물의 수성 현탁액에 다른 생물의 노출을하고있었습니다. 그러나 독성 분석은 잠재적 인 포식자가 자연 상태에서 먹이를 인식하는 방식을 모방하지 않으며, 후속 연구는 독성과 기호성 12 ~ 13 사이의 관계를 발견했다. 그것은 권위있는 저널에 출판이 거의 또는 전혀 ecologica을 가진 기술을 사용하는 것은 놀라운 일이다L 관련성 14-15 것을이 연구가 아직 널리 인용된다 오늘. 그것은 독성 데이터를 기반으로 연구 16-18을 게시 할 계속주의하는 것이 더욱 놀랍다. 여기에 설명 된 생물 검정 방법은 antipredatory 화학 방어를 평가하기 위해 해양 화학 생태에 대한 생태 학적으로 중요한 접근 방법을 제공하기 위해 1980 년대 후반에 개발되었다. 방법은 생태 학적으로 의미있는 독성 데이터보다 기호성 데이터를 제공, 식품 영양 학적으로 유사한 매트릭스 천연 농도 대상 유기체로부터 조 유기 추출물을 샘플링 포식자 모델을 필요로한다. (1) 적절한 제너럴 육식 동물이 먹이 분석에 사용되어야한다, (2) 모든 극성 유기 대사가 철저의 조직에서 추출해야합니다 해양 생물의 조직의 antipredatory 활동을 평가하는 일반적인 방법은 네 가지 중요한 기준을 포함 생물체를 대상으로, (3) 대사 ㄴ해야같은 체적 농도 영양 적절한 실험 음식에 혼합 전자는 그들이 추출하고, (4) 실험 설계 및 통계 방법은 의미있는 메트릭이 상대 distastefulness을 표시하기 위해 제공해야하는 유기체에서 발견. 아래에 설명 된 절차는 카리브해 해양 무척추 동물에 antipredatory 화학 방어를 평가하기 위해 설계되었습니다. 이 종은 카리브해 산호초에 공통이며, 저서 무척추 동물 (19)의 넓은 구색을 샘플링 알려져 있기 때문에 우리는 모델 육식 물고기 bluehead 놀래기과의 바닷물 고기, Thalassoma의 bifasciatum을 사용합니다. 표적 조직은 유기체로부터 먼저 추출 식품 혼합물과 결합하고, 마지막 T. 그룹에 제공되며 bifasciatum은 추출물 처리 된 음식을 거부 여부를 관찰합니다. 이 방법을 사용하여 분석 데이터는 해양 생물 12,20-21, L의 수비 화학에 중요한 통찰력을 제공하고 있습니다IFE의 역사 절충 22-24, 지역 사회 생태 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.

여기에 우리가 일반적으로 카리브해 스폰지의 여섯 종 (그림 2)이 생물 검정의 결과를보고합니다. 이러한 데이터는 처음 Pawlik 등. (12)에 의해 1995 년에 출판 및 공동 발생하는 분류군 중 화학 방어 전략의 차이를 조사하기 위해이 방식의 힘을 보여되었다. 결과는 표준 오차 (SE) 각 종 + 먹은 음식 펠릿의 평균 숫자로보고되었다. 거의 펠릿 Agelas의 clathrodes, Amphimedon compre...

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

이러한 생물학적 검정은 식품 영양 학적으로 유사한 매트릭스를 이용 농도 천연 해양 생물의 조직 유기 추출물로부터 병입 억지 대사의 존재를 평가하기위한 모델 육 물고기를 이용한다.

물고기 먹이 실험 생물 검정은 해양 생물의 조직에서 이차 대사 산물의 Antipredatory 활동을 평가하기 위해

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

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