作者感谢皮分林小姐、魏隆先生、裴小姐和佳禄博士在实地抽样和实验室分析中对 "CATNET" 的实际开发和应用进行了协助。在取样远征期间, 船员和技术员对船上研究船海洋研究和海洋研究 II 的协助非常感谢。这项工作部分是由台湾科学技术部资助的91-2611 米-002-007、95-2611 米-002-009、96-2611 米-002-004、97-3114 米-002-006、104-2611 米-002-019。这篇手稿是为了纪念翬小姐对台湾海洋研究的巨大奉献和贡献而写的。

以下议定书涉及使用有害化学品。请仔细阅读安全数据表 (SDS), 并遵循机构化学安全指南。
1. 多层重力过滤取样器的制备
<ol><li>
取样器清洗
	<ol>
<li>用 1% (w/v) 的阴离子蛋白酶酶洗涤剂溶液填充油管和过滤单元, 浸泡24小时, 用反渗透双蒸馏水 (RO-DDW) 彻底冲洗多层重力过滤取样器, 然后将其填充 0.1% (v/v)盐酸 (盐酸, 试剂级) 和浸泡72小时。</li>
		<li>用反渗透双蒸馏去离子水 (RO-DIW) 将多层重力过滤取样器彻底冲洗三至五次, 每次至少20公升, 并将组合存放在塑料袋中。</li>
	</ol>
</li>
	<li>
微粒样品容器清洁/准备
	<ol>
<li>使用低密度聚乙烯 (聚乙烯, 125 毫升) 或氟化乙烯丙烯 (FEP, 125 毫升) 瓶作为容器的微粒。先用碱性洗涤剂 (微, 1%), 然后在 50% (v/v) 硝酸 (HNO3, 试剂级), 然后 10% (v/v) HCl 溶液的至少 24, 48 和 24 h, 清洗瓶子, 分别。在两个浸泡步骤之间用脱电离水 (RO-DIW) 冲洗瓶子。</li>
		<li>在最后的 HCl 浸泡后, 彻底冲洗瓶子与脱电离水 (RO-DIW), 并干燥的瓶子在一个干净的房间或 class-100 干净的长凳。 将清洗过的瓶子连接到多层重力过滤取样器, 或将清洗过的瓶子密封在 PE 拉链袋中, 并用双袋包装运输。</li>
	</ol>
</li>
	<li>
多层重力过滤取样器的组合
	<ol>
<li>将六4米长的耐化学热塑性弹性体管 (外径0.635 厘米) 连接到取样器顶部的六方向入口。</li>
		<li>将三种不同的网状尼龙过滤器与低密度聚乙烯样品容器 (125 毫升聚乙烯) 依次组装在洁净室 (工作台) 清洗后 (见下文), 用10µm 网格过滤器定位在外部, 63 µm 网格过滤器在中间, 和153µm 网格过滤器在里面。运输时, 将多层重力过滤取样器存储在两层聚乙烯 (PE) 袋中, 然后放入聚丙烯 (PP) 装运容器中。</li>
	</ol>
</li>
</ol>2. 取样
<ol><li>
示例集合
	<ol>
<li>抵达取样地点后, 有一人从研究船甲板上的装运集装箱中取出多层重力过滤取样器, 并用取样装置打开袋子。然后, 让他们穿上 PE 手套, 将六4米热塑性弹性体管连接到高架多瓶取样阵列上的六 20 L 聚四氟乙烯涂层取样瓶的水闸门, 并将海水引导到这个过滤单元中。海水将流经定向入口, 粒子/浮游生物将被轻轻地分离/分割通过网, 并落户到125毫升的低密度聚乙烯瓶, 在网底固定。</li>
		<li>海水流经后 (通常为120升为沿海海水, 240 升为开阔海水), 按顺序去除每个网 (首先, 153 µm, 然后63µm, 最后10µm) 在一个 class-100 干净的工作台, 然后喷洒净与微量金属清洁0.4 µm filtered 海水冲洗出网内表面的任何浮游生物。在125毫升聚乙烯瓶中收集浓缩颗粒/浮游生物的海水。</li>
		<li>将这些瓶子从网中拧开, 再通过酸性洗涤的真空过滤装置, 在低真空下用预称量、酸洗47毫米、10µm 孔径聚碳酸酯过滤器过滤溶液。条件 (< 5 帕)。</li>
		<li>要收集小于10µm 的微粒/浮游生物, 请等待至少20升的海水流经取样器, 然后在 5 L PE 容器中收集两到五升的水, 然后用酸性洗涤的真空过滤装置过滤这些水样, 并预重, 酸性水洗, 47 毫米, 0.4 µm 孔径聚碳酸酯过滤器。</li>
		<li>真空过滤后, 立即用高纯度的 DDW 水冲洗样品过滤器, 去除海水中的残留物, 尽量减少海水盐分对测定颗粒/浮游生物干重的影响。保持冲洗量只有几毫升, 以防止破坏脆弱的浮游生物。</li>
		<li>然后, 在此冲洗步骤后, 小心地将过滤器从真空过滤单元中取出, 将样品过滤器储存在酸洗、预压丙烯酸塑料培养皿中, 并密封在密封塑料袋中。把袋子放在-20 ˚C 的冰箱里, 直到回到陆地实验室进行进一步的样品预处理和化学分析。</li>
	</ol>
</li>
</ol>3. 样品处理
<ol><li>
颗粒的冷冻干燥和消化
	<ol>
<li>将过滤器与颗粒样品放在冷冻干燥机的收集室中, 并打开机器。当机器温度达到-40 摄氏度时, 打开机器的真空泵, 开始冷冻干燥过程。 注: 真空度应保持稳定在0.12 毫巴以下。请仔细阅读用户手册, 并按照制造商的指导方针为每个步骤。</li>
		<li>72小时后, 关闭冷冻干燥机, 取出干燥过滤器并称重。然后, 将干样过滤器放入预称量的 perfluoroalkoxy 烷烃 (粉煤灰) 容器 (60 毫升容量), 并将3毫升浓缩的超纯硝酸加入容器中2,3,6,7。</li>
		<li>用扭矩扳手将容器拧紧至2.5 公斤的恒定扭矩, 并将容器放在一个常规烤箱中, 在130摄氏度为12小时, 用于第一消化序列。冷却后, 从烤箱中取出容器, 打开容器, 将2毫升的超纯氢氟酸加入容器中2,3,6,7。</li>
		<li>拧紧的扭矩为2.5 公斤的船只, 并把这些船只在一个传统的烤箱在130°c 12 小时, 这是第二消化序列。冷却后, 打开容器, 并将16毫升4.5% 超纯硼酸溶液添加到容器中2,3,6,7。</li>
		<li>将容器拧紧至2.5 公斤米的恒定扭矩, 并在烤箱中以130°c 的样品消化, 最后消化顺序为12小时。冷却后, 权衡每一个容器, 并确定每一个消化溶液的最终质量和特定质量, 以产生最终的配药体积。 注意: 具体质量是通过测量准确的1.00 毫升配药的重量来确定的。</li>
		<li>小心倒入30毫升酸清洗 PE 瓶配药进一步的微量金属分析。</li>
	</ol>
</li>
	<li>
微量金属分析
	<ol>
<li>用石墨炉原子吸收光谱仪 (GF-原子吸收光谱法)6测定颗粒的消化溶液中的微量金属浓度 (镉、铜、铁、锰、镍和铝)。</li>
		<li>作为精度测试, 使用经认证的参考材料 (CRM), 如加拿大国家研究委员会的海洋沉积物参考材料, 河口泥沙标准参考材料从国家标准和技术研究所美国和浮游生物参考材料从欧洲委员会的科学和知识服务。该过程使 CRM 中提供的微量金属的认证价值得到95% 到107% 的恢复。</li>
	</ol>
</li>
</ol>

<ol>
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Oceanogr. 133, 32-42 (2015).</li><li>Collier, R., Edmond, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+trace+element+geochemistry+of+marine+biogenic+particulate+matter.">The trace element geochemistry of marine biogenic particulate matter.</a> Progr. Oceanogr. 13, 113-199 (1984).</li><li>Donat, J. R., Bruland, K. W., Steinnes, E., Salbu, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+elements+in+the+oceans.">Trace elements in the oceans.</a> Trace Elements in Natural Waters. , 247-280 (1995).</li><li>Wen, L. -. S., Santschi, P., Tang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction+between+radioactively+labeled+colloids+and+natural+particles:+evidence+for+colloidal+pumping.">Interaction between radioactively labeled colloids and natural particles: evidence for colloidal pumping.</a> Geochim. Cosmochim. Ac. 61, 2867-2878 (1997).</li><li>Wen, L. -. S., Warnken, K., Santschi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+organic+carbon,+iron,+and+aluminium+oxyhydroxides+as+trace+metal+carriers:+Comparison+between+the+Trinity+River+and+the+Trinity+River+Estuary+(Galveston+Bay,+Texas).">The role of organic carbon, iron, and aluminium oxyhydroxides as trace metal carriers: Comparison between the Trinity River and the Trinity River Estuary (Galveston Bay, Texas).</a> Mar. 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Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+elemental+composition+of+some+marine+phytoplankton.">The elemental composition of some marine phytoplankton.</a> J. Phycol. 39, 1145-1159 (2003).</li><li>McDonnell, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+oceanographic+toolbox+for+the+collection+of+sinking+and+suspended+marine+particles.">The oceanographic toolbox for the collection of sinking and suspended marine particles.</a> Prog. Oceanogr. 133, 17-31 (2015).</li><li>Wen, L. -. S., Li, W. -. H., Zhuang, G. -. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Multiple layer filtering and collecting device. , (2005).</li><li>Ho, T. -. Y., Wen, L. -. S., You, C. -. 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Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+metal+composition+of+size-fractionated+plankton+in+the+Western+Philippine+Sea:+the+impact+of+anthropogenic+aerosol+deposition.">Trace metal composition of size-fractionated plankton in the Western Philippine Sea: the impact of anthropogenic aerosol deposition.</a> Limnol Oceanogr. , (2017).</li><li>Grasshoff, K., Kremling, K., Ehrhardt, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods of seawater analysis. , (2007).</li></ol>

合著者艾伦. 创先生是该公司的专利共同所有者和总经理 (中信仪器有限公司), 为感兴趣的用户制造了这个收集设备。该专利于 5月9日 (th, 201512) 结束。

获得可靠的微量金属浓度的浮游生物和悬浮粒子在自然水域, 通常是在非常低浓度, 需要很大的照顾, 在样品收集, 处理, 预处理和分析, 目的是减少污染。因此, 设计和准备取样齿轮、样品容器和用于收集和处理样品的材料的程序是在海洋环境中获得高质量的微量金属数据的关键步骤。随着近几十年来新的粒子采集方法的发展, 我们对粒子动力学和微量元素地球化学的认识也在不断拓宽。本文介绍了...

海洋中的粒子在海洋生物地球化学循环中起着重要作用1。粒子的大部分属性, 如大小、矿物学和组成, 都可以从一个地质或水文设置到另一个2。此外, 海洋中元素的分布也与海洋浮游植物的生命周期有关: 生长、死亡、下沉和再矿化3,4。海洋颗粒的大小至少有4级, 从亚微米颗粒到大骨料 (> 5 毫米) 不等。大多数粒子都是从生物学上衍生出来的, 从病毒裂解、渗出、分泌、粪便颗粒的产生、等等。其他粒子是由细胞、细胞碎片或胆汁成石材料的物理凝固而形成的1。微粒的各种各样的化学和生物特征控制地球化学循环和生物过程发生在和在微粒之内4,5,6。这些微粒是重要栖所和食物来源为一些有机体, 例如浮游动物或 saprotrophs。因此, 粒子的命运常常与它们的大小有关, 而粒子的生物过程可以改变它们的尺寸。
取样海洋微粒通常需要过滤, 但这种方法在识别粒子的性质方面引入了一定的模糊性, 因为海洋颗粒在成分和大小上不均匀。悬浮粒子, 主要由几乎永久悬浮的小颗粒和低密度粒子组成, 在悬浮液中, 由于水动力条件, 在短时间内混合了不同数量的大颗粒和高密度粒子。7. 在研究船8上, 通过浮游生物拖曳或悬浮浮游生物网收集浮游生物样品的微量金属组成的第一个报告。作者经常在样品中发现金属微粒和油漆芯片, 这表明在化学分析中, 海洋微粒取样过程中存在严重的污染问题。其他努力包括用橡皮筏进行净拖曳或使用聚氯乙烯 (PVC) 手绞车3。对粒子进行可靠取样的困难使得我们对海洋粒子化学成分的理解更加困难, 特别是对于微量元素。因此, 关于浮游植物中微量元素浓度的最关键信息来自于文化研究9,10。这一认识促使海洋科学家在过去三十年中为研究海洋中的粒子创造了新的方法11。
海洋学家使用了各种取样技术, 包括船用过滤、原位过滤和沉积陷阱11。处理大量的海水来收集无污染的样品可能是一个挑战, 特别是对于开阔的海洋和深水域, 其中颗粒浓度非常低 (0.001-0.1 毫克/升)。还需要过滤大量的海水, 以获得足够数量的微粒来测量微量金属浓度。一些研究人员使用了粒度分馏法将悬浮粒子与下沉粒子分离。然而, 颗粒的大小、孔隙度、密度和形状都能影响颗粒的下沉速度。沉淀物陷阱不是收集悬浮粒子的实用工具, 因为这些是为下沉粒子设计的。因此, 有必要制定取样和处理方法, 以便收集足够数量的悬浮粒子, 并将其污染降到最低。因此, 在海洋学家的取样工具箱中, 通过原位过滤来进行尺寸分割仍然是一个有希望的工具, 因为它可以揭示海洋粒子动力学的关键信息。在这里, 我们描述了一个成功测试的微量金属清洁, 多层重力过滤取样装置, 它可以处理大容量 (120-240 升) 的船上的海水在一个通行证从聚四氟乙烯 (聚四氟乙烯) 涂层水取样瓶在一个多瓶取样阵列。该取样装置采用酸性水洗合成尼龙网, 并将网套在聚碳酸酯容器内, 以轻轻收集大小分级悬浮物质和浮游植物12,13, 14,15 (图 1)。这项工作的目的是提供一个更好的工具, 以研究在海洋环境中的金属粒子协会及其反应动力学, 并提高我们对各种浮游生物, 粒子和微量金属的命运的理解, 在这些环境。

<table>
	<tbody>
		<tr>
			<td>thermoplastic elastomer (C-Flex) Tubings</td>
			<td>Cole Palmer</td>
			<td>EW-06424-67</td>
			<td>O.D. 0.635 cm, Opaque White 1/8&#34;ID x 1/4&#34;OD, 25 ft/pack</td>
		</tr>
		<tr>
			<td>LDPE Bottle (Nalgene)</td>
			<td>ThermoFisher Scientific</td>
			<td>2103-0004</td>
			<td>125 mL, Nalgene Wide-Mouth LDPE Bottles with Closure</td>
		</tr>
		<tr>
			<td>anionic protease enzyme detergent detergent (Tergazyme)</td>
			<td>Alconox</td>
			<td>1104-1</td>
			<td>1&#215;4 lb box (1.8 kg)</td>
		</tr>
		<tr>
			<td>Hydrochloric Acid</td>
			<td>Sigma-Aldrich</td>
			<td>258148</td>
			<td>Reagent grade</td>
		</tr>
		<tr>
			<td>Nitric acid</td>
			<td>Sigma-Aldrich</td>
			<td>695025</td>
			<td>Reagent grade</td>
		</tr>
		<tr>
			<td>alkaline detergnet (Micro)</td>
			<td>Cole Palmer</td>
			<td>EW-99999-14</td>
			<td>Micro-90 Cleaning Solution</td>
		</tr>
		<tr>
			<td>polycarbonate filter, 47 mm, 0.4 &#181;m</td>
			<td>Sigma-Aldrich</td>
			<td>WHA111107</td>
			<td>Whatman Nuclepore Track-Etched Membranes, diam. 47 mm, pore size 0.4 &#956;m, polycarbonate</td>
		</tr>
		<tr>
			<td>polycarbonate filter, 47 mm, 10 &#181;m</td>
			<td>Sigma-Aldrich</td>
			<td>WHA111115</td>
			<td>Whatman Nuclepore Track-Etched Membranes, diam. 47 mm, pore size 10 &#956;m, polycarbonate</td>
		</tr>
		<tr>
			<td>PFA vessel, 60 ml capacity</td>
			<td>Savillex</td>
			<td>300-060-03</td>
			<td>60 mL Digestion Vessel, Flat Interior, Flat Exterior, Buttress Threaded Top</td>
		</tr>
		<tr>
			<td>Nitric acid, ultrapure</td>
			<td>Seastar Chemicals</td>
			<td>N/A</td>
			<td>BASELINE Nitric Acid</td>
		</tr>
		<tr>
			<td>HF, ultrapure</td>
			<td>Seastar Chemicals</td>
			<td>N/A</td>
			<td>BASELINE Hydrofluoric Acid</td>
		</tr>
		<tr>
			<td>Boric acid, ultrapure</td>
			<td>Seastar Chemicals</td>
			<td>N/A</td>
			<td>BASELINE Hydrobromic Acid</td>
		</tr>
		<tr>
			<td>polyethylene (PE) gloves</td>
			<td>Safty Zone</td>
			<td>GDPL-MD-5</td>
			<td>Clear Powder Free Polyethylene Gloves</td>
		</tr>
		<tr>
			<td>Multiple layer filtering and collecting device</td>
			<td>Sino Instrumnets Co. Ltd</td>
			<td>not available</td>
			<td>Multiple layer filtering and collecting device, CATNET</td>
		</tr>
		<tr>
			<td>10 um Nylon filters, Nitex</td>
			<td>Dynamic Aqua-Supply Ltd.</td>
			<td>NTX 10</td>
			<td>Nitex - Standard Widths (40 - 44 inches)</td>
		</tr>
		<tr>
			<td>60 um Nylon filters, Nitex</td>
			<td>Dynamic Aqua-Supply Ltd.</td>
			<td>NTX 60</td>
			<td>Nitex - Standard Widths (40 - 44 inches)</td>
		</tr>
		<tr>
			<td>150 um Nylon filters, Nitex</td>
			<td>Dynamic Aqua-Supply Ltd.</td>
			<td>NTX 150</td>
			<td>Nitex - Standard Widths (40 - 44 inches)</td>
		</tr>
		<tr>
			<td>torque wrench</td>
			<td>Halfords</td>
			<td>200238</td>
			<td>Halfords Professional Torque Wrench 8-60Nm</td>
		</tr>
		<tr>
			<td>multi-bottle sampling array, Rosette</td>
			<td>General Oceanics</td>
			<td>Model 1018</td>
			<td>Rosette Sampler</td>
		</tr>
		<tr>
			<td>PTFE-coated sampling bottles, GO-Flo</td>
			<td>General Oceanics</td>
			<td>108020T</td>
			<td>GO-Flo water sampler teflon coated</td>
		</tr>
		<tr>
			<td>Marine sediment reference materials</td>
			<td>National Research Council Canada</td>
			<td>MESS-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Estuarine sediment standard reference material</td>
			<td>National Institute of Standards and Technology</td>
			<td>1646a</td>
			<td></td>
		</tr>
		<tr>
			<td>Plankton reference material</td>
			<td>The European Commission&#39;s science and knowledge service</td>
			<td>CRM414</td>
			<td></td>
		</tr>
	</tbody>
</table>

an ultra-clean multilayer apparatus for collecting size fractionated marine plankton and suspended particles

海洋中许多微量元素的分布与海洋浮游生物的生长、死亡和再矿化以及悬浮/下沉粒子的沉积密切相关。在这里, 我们提出了一个全塑料 (聚丙烯和聚碳酸酯), 多层过滤系统收集悬浮微粒物质 (SPM) 在海上。这种超清洁取样装置是专门为微量元素研究而设计和开发的。对所有非金属材料进行细致的选择, 并利用在线流动过程, 最大限度地减少取样过程中可能发生的金属污染。该系统已成功测试和调整, 以确定痕量金属 (如, 铁, 铝, 锰, 镉, 铜, 镍) 对不同大小的颗粒在沿海和开阔海域。东南亚时间序列 (座位) 站南中国海的结果表明, 透光地区浮游生物的日变化和空间分布可以很容易地得到解决和识别。台湾海峡地表水中颗粒粒度的化学分析表明, 较大的颗粒 (> 153 µm) 主要是生物衍生的, 而较小的颗粒 (10-63 µm) 主要由无机物质组成。除 Cd 外, 金属 (铁、铝、锰、铜、镍) 的浓度随尺寸的增大而减小。

随着现代海洋学的发展, 使用 "清洁技术" 获得准确的海洋颗粒或浮游生物中微量金属浓度是目前的普遍做法。由于天然水体中的大多数粒子都处于低镁/升到µg 的范围内, 因此必须对大量海水进行处理, 以调查在环境环境中微量金属对各种粒子的地球化学和生物效应。使用清洁、多层重力过滤 ("CATNET") 取样技术 (图 1), 在确定使用常规加压死端过滤和 CA...

Watch this Scientific Journal Video about An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles at JoVE.com

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

海洋浮游生物和悬浮颗粒的超洁净多层采集装置

浮游生物和悬浮粒子在海洋生物地球化学循环中起着重要作用。在这里, 我们提供了一种超清洁, 低应力的方法, 以收集各种大小的颗粒和浮游生物在海上的能力, 处理大量的海水。

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

an-ultra-clean-multilayer-apparatus-for-collecting-size-fractionated

Research

JoVE Journal

Environment

8.7K 次观看.  National Taiwan University. 浮游生物和悬浮粒子在海洋生物地球化学循环中起着重要作用。在这里, 我们提供了一种超清洁, 低应力的方法, 以收集各种大小的颗粒和浮游生物在海上的能力, 处理大量的海水。

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

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.

This bioassay employs a model predatory fish to assess the presence of feeding-deterrent metabolites from organic extracts of the tissues of marine organisms at natural concentrations using a nutritionally comparable food matrix.

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 ...

科研

环境科学

A Protocol for Collecting and Constructing Soil Core Lysimeters

Leaching of nutrients from land applied fertilizers and manure used in agriculture can lead to accelerated eutrophication of surface water. Because the landscape has complex and varied soil morphology, an accompanying disparity in flow paths for leachate through the soil macropore and matrix structure is present. The rate of flow through these paths is further affected by antecedent soil moisture. Lysimeters are used to quantify flow rate, volume of water and concentration of nutrients leaching downward through soils. While many lysimeter designs exist, accurately determining the volume of water and mass balance of nutrients is best accomplished with bounded lysimeters that leave the natural soil structure intact. 
Here we present a detailed method for the extraction and construction of soil core lysimeters equipped with soil moisture sensors at 5 cm and 25 cm depths. Lysimeters from four different Coastal Plain soils (Bojac, Evesboro, Quindocqua and Sassafras) were collected on the Delmarva Peninsula and moved to an indoor climate controlled facility. Soils were irrigated once weekly with the equivalent of 2 cm of rainfall to draw down soil nitrate-N concentrations. At the end of the draw down period, poultry litter was applied (162 kg TN ha-1) and leaching was resumed for an additional five weeks. Total recovery of applied irrigation water varied from 71% to 85%. Nitrate-N concentration varied over the course of the study from an average of 27.1 mg L-1 before litter application to 40.3 mg L-1 following litter application. While greatest flux of nutrients was measured in soils dominated by coarse sand (Sassafras) the greatest immediate flux occurred from the finest textured soil with pronounced macropore development (Quindocqua).

A detailed method for extraction and assembly of intact soil core lysimeters and their use for study of leachate and associated loss of nutrients from surface applied poultry litter is demonstrated.

一种用于收集和构建土芯协议测渗计

Leaching of nutrients from land applied fertilizers and manure used in agriculture can lead to accelerated eutrophication of surface water. Because the ...

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.

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，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 ...

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

对水生生命值评价悬浮泥沙影响的流通系统曝光

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of scientific publications address microplastic pollution of the sea surface. The protocol below describes the methodology for sampling, sample preparation, separation and chemical identification of microplastic particles. A manta net fixed on an &#187;A frame&#171; attached to the side of the vessel was used for sampling. Microplastic particles caught in the cod end of the net were separated from samples by visual identification and use of stereomicroscopes. Particles were analyzed for their size using an image analysis program and for their chemical structure using ATR-FTIR and micro FTIR spectroscopy. The described protocol is in line with recommendations for microplastics monitoring published by the Marine Strategy Framework Directive (MSFD) Technical Subgroup on Marine Litter. This written protocol with video guide will support the work of researchers that deal with microplastics monitoring all over the world.

The protocol below describes the methodology for: microplastics sampling on the sea surface, separation of microplastic and chemical identification of particles. This protocol is in line with the recommendations for microplastics monitoring published by the MSFD Technical Subgroup on Marine Litter.

协议塑料微粒采样的海面和样品分析

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment during manufacturing and transport. Because of their environmental persistence, they are widely distributed in the oceans and on beaches all over the world. They can act as a vector of potentially toxic organic compounds (e.g., polychlorinated biphenyls) and might consequently negatively affect marine organisms. Their possible impacts along the food chain are not yet well understood. In order to assess the hazards associated with the occurrence of plastic pellets in the marine environment, it is necessary to develop methodologies that allow for rapid determination of associated organic contaminant levels. The present protocol describes the different steps required for sampling resin pellets, analyzing adsorbed organochlorine pesticides (OCPs) and identifying the plastic type. The focus is on the extraction of OCPs from plastic pellets by means of a pressurized fluid extractor (PFE) and on the polymer chemical analysis applying Fourier Transform-InfraRed (FT-IR) spectroscopy. The developed methodology focuses on 11 OCPs and related compounds, including dichlorodiphenyltrichloroethane (DDT) and its two main metabolites, lindane and two production isomers, as well as the two biologically active isomers of technical endosulfan. This protocol constitutes a simple and rapid alternative to existing methodology for evaluating the concentration of organic contaminants adsorbed on plastic pieces.

Microplastics act as vector of potentially toxic organic contaminants with unpredictable effects. This protocol describes an alternative methodology for assessing the levels of organochlorine pesticides adsorbed on plastic pellets and identifying the polymer chemical structure. The focus is on pressurized fluid extraction and attenuated total reflectance Fourier transform infrared spectroscopy.

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 ...

Preparation of Authigenic Pyrite from Methane-bearing Sediments for In Situ Sulfur Isotope Analysis Using SIMS

Different sulfur isotope compositions of authigenic pyrite typically result from the sulfate-driven anaerobic oxidation of methane (SO4-AOM) and organiclastic sulfate reduction (OSR) in marine sediments. However, unravelling the complex pyritization sequence is a challenge because of the coexistence of different sequentially formed pyrite phases. This manuscript describes a sample preparation procedure that enables the use of secondary ion mass spectroscopy (SIMS) to obtain in situ &#948;34S values of various pyrite generations. This allows researchers to constrain how SO4-AOM affects pyritization in methane-bearing sediments. SIMS analysis revealed an extreme range in &#948;34S values, spanning from -41.6 to +114.8&#8240;, which is much wider than the range of &#948;34S values obtained by the traditional bulk sulfur isotope analysis of the same samples. Pyrite in the shallow sediment mainly consists of 34S-depleted framboids, suggesting early diagenetic formation by OSR. Deeper in the sediment, more pyrite occurs as overgrowths and euhedral crystals, which display much higher SIMS &#948;34S values than the framboids. Such 34S-enriched pyrite is related to enhanced SO4-AOM at the sulfate-methane transition zone, postdating OSR. High-resolution in situ SIMS sulfur isotope analyses allow for the reconstruction of the pyritization processes, which cannot be resolved by bulk sulfur isotope analysis.

Preparation of Authigenic Pyrite from Methane-bearing Sediments for In Situ Sulfur Isotope Analysis Using SIMS

Analyses of the sulfur isotopic composition (&#948;34S) of pyrite from methane-bearing sediments have typically focused on bulk samples. Here, we applied secondary ion mass spectroscopy to analyze the &#948;34S values of various pyrite generations to understand the diagenetic history of pyritization.

原位含甲烷沉积物中自黄铁矿的制备及应用模拟器硫同位素分析

Different sulfur isotope compositions of authigenic pyrite typically result from the sulfate-driven anaerobic oxidation of methane (SO4-AOM) and ...

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

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 estimating fish abundance. We developed and implemented a rotating stereo-video camera tool that covers a full 360 degrees of sampling, which maximizes sampling effort compared to stationary camera tools. A variety of studies have detailed the ability of static, stereo-camera systems to obtain highly accurate and precise measurements of fish; the focus here was on the development of methodological approaches to quantify fish density using rotating camera systems. The first approach was to develop a modification of the metric MaxN, which typically is a conservative count of the minimum number of fish observed on a given camera survey. We redefine MaxN to be the maximum number of fish observed in any given rotation of the camera system. When precautions are taken to avoid double counting, this method for MaxN may more accurately reflect true abundance than that obtained from a fixed camera. Secondly, because stereo-video allows fish to be mapped in three-dimensional space, precise estimates of the distance-from-camera can be obtained for each fish. By using the 95% percentile of the observed distance from camera to establish species-specific areas surveyed, we account for differences in detectability among species while avoiding diluting density estimates by using the maximum distance a species was observed. Accounting for this range of detectability is critical to accurately estimate fish abundances. This methodology will facilitate the integration of rotating stereo-video tools in both applied science and management contexts.

We describe a new method for counting fishes, and estimating relative abundance (MaxN) and fish density using rotating stereo-video camera systems. We also demonstrate how to use distance from camera (Z distance) to estimate species-specific detectability.

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 ...

Methods for Image-based Surveys of Benthic Macroinvertebrates and Their Habitat Exemplified by the Drop Camera Survey for the Atlantic Sea Scallop

Underwater imaging has long been used in the field of marine ecology but decreasing costs of high-resolution cameras and data storage have made the approach more practical than in the past. Image-based surveys allow for initial samples to be revisited and are non-invasive compared to traditional survey methods that typically involve nets or dredges. Protocols for image-based surveys can vary greatly but should be driven by target species behavior and survey objectives. To demonstrate this, we describe our most recent methods for an Atlantic sea scallop (Placopecten magellanicus) drop camera survey to provide a procedural example and representative results. The procedure is divided into three critical steps that include survey design, data collection, and data products. The influence of scallop behavior and the survey goal of providing an independent assessment of the U.S. sea scallop resource on the survey procedure are then discussed in the context of generalizing the method. Overall, the broad applicability and flexibility of the University of Massachusetts Dartmouth School for Marine Science and Technology (SMAST) drop camera survey demonstrates the method could be generalized and applied to a variety of sessile invertebrates or habitat focused research.

Image based surveying is an increasingly practical, non-invasive method to sample the marine environment. We present the protocol of a drop camera survey that estimates the abundance and distribution of the Atlantic sea scallop (Placopecten magellanicus). We discuss how this protocol can be generalized for application to other benthic macroinvertebrates.

基于图像的海底无脊椎动物及其栖息地调查方法大西洋扇贝落相机调查

Underwater imaging has long been used in the field of marine ecology but decreasing costs of high-resolution cameras and data storage have made the ...

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

Barnacles are marine crustaceans with a sessile adult and free-swimming, planktonic larvae. The barnacle Balanus (Amphibalanus) improvisus is particularly relevant as a model for the studies of osmoregulatory mechanisms because of its extreme tolerance to low salinity. It is also widely used as a model of settling biology, in particular in relation to antifouling research. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae for studies. A protocol for the all-year-round culturing of B. improvisus has been developed and a detailed description of all steps in the production line is outlined (i.e., the establishment of adult cultures on panels, the collection and rearing of barnacle larvae, and the administration of feed for adults and larvae). The description also provides guidance on troubleshooting and discusses critical parameters (e.g., the removal of contamination, the production of high-quality feed, the manpower needed, and the importance of high-quality seawater). Each batch from the culturing system maximally yields roughly 12,000 nauplii and can deliver four batches in a week, so up to almost 50,000 larvae per week can be produced. The method used to culture B. improvisus is, probably, to a large extent also applicable to other marine invertebrates with free-swimminglarvae. Protocols are presented for the dissection of various tissues from adults as well as the production of high-quality RNA for studies on gene expression. It is also described how cultured adults and reared cyprids can be utilized in a wide array of experimental designs for examining gene expression in relation to external factors. The use of cultured barnacles in gene expression is illustrated with studies of possible osmoregulatory roles of Na+/K+ ATPase and aquaporins.

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

The barnacle Balanus (Amphibalanus) improvisus is a model for studying osmoregulation and antifouling. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae. Here, a protocol for the all-year-round culturing of B. improvisus is described, including the production of larvae. The use of cultured barnacles in gene expression studies is illustrated.

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 ...