提交人要感谢 NEFSC 和 NOAA 水产养殖服务处提供资金。作者感谢他们的学术和工业合作伙伴, 斯科特 Lindell, 伍兹洞海洋学研究所的研究专家, 和卡特琳娜海洋牧场的首席执行官菲尔 Cruver, 他安排和提供进入近海贻贝生长地区的机会。如果没有下列工作平台, 这项工作就不可能实现;r/v船长杰克拥有由海洋生物实验室拥有和管理的卡特琳娜海牧场, 和Loosanoff 维克多渔业, 东北渔业科学中心经营的 r/v。我们还感谢船长吉姆 Cvitanovich 和比尔. 克林的专长。沃纳 Schreiner 提供了他的技术专长, 设计和制造框架, 框架表和压载舱, 头水箱, 和实验室。

1. 框架工作台和加料装置
<ol><li>构造和装配框架表, 包括两个框架, 一个框架表, 和一个压载坦克, 如图 1a所示。<ol>
<li>建立最外层的框架130厘米长, 92 厘米宽, 90 厘米高使用 0.65 cm 聚氯乙烯 (PVC) 股票。使用不锈钢螺母和螺栓形成框架。</li>
		<li>从4厘米 x 10 厘米聚氯乙烯 (PVC) 库存中建造最里面的框架 (长125厘米和80厘米宽)。适合在框架的短边的顶部加强的部分接受内部框架框架。永久修复不锈钢针脚, 使内部框架在外部框架内自由摆动。</li>
		<li>同样, 在内框的长边包括钢筋截面, 以容纳安装在框架表中的不锈钢销, 使其自由摆动。</li>
		<li>用可拆卸的镇流器将 PVC 立方体储存起来。将压载液舱装满85公斤海水, 并将50公斤锌重量插入压载液舱底部; 及它作为一个平衡, 以抑制, 但不限制, 摆动的表。 注: 压载液舱由不锈钢螺母和螺栓连接到框架表。</li>
	</ol>
</li>
</ol><img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58213/58213fig1.jpg"> 图 1: 在船上使用 biodeposition 方法为定量双壳悬浮饲料而研制的框架表和加料装置.(a) 此面板显示了带有加料装置的装配式框架表的图像。(b) 此面板显示了装配式送料装置的示意图。<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/58213/58213fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="2"><li>构造和装配送料装置, 由头水箱和2套10进料室组成 (图 1b)。<ol>
<li>用6.5 毫米 PVC 建造头部水箱, 长度 x 30 厘米, 高度为70厘米 (图 2a)。钻一个25毫米直径的孔在中心的左30厘米一侧3厘米从顶部。</li>
		<li>钻10孔13毫米直径通过每一个70厘米 PVC 件的矩形, 使每个孔的中心是2.5 厘米从基地。从头水箱一侧钻40毫米的第一孔;然后, 连续孔的中心相距69毫米。</li>
		<li>将塑料隔板连接器放置在每个孔内直径为7毫米的内螺纹, 以允许水离开头水箱。将6.5 毫米内径的硅管安装在接头上。在每管中间, 在头水箱和进料室之间, 将可调节阀与油管连接, 以控制进入进料室的流量。 注: 为确保颗粒保持在头水箱水中悬浮, 均匀分布在加料室内, 使用空气石或空气管在整个油箱中添加曝气。</li>
		<li>每个进给室的内部测量值为17.5 厘米, 长度 x 6 厘米, 宽度 x 6 厘米 (图 2b)。钻一个13毫米直径的孔在6厘米的一侧的中心, 使孔的中心是15毫米从底部。在每个腔的相对6厘米的一侧, 从底部钻一个13毫米直径的孔45毫米。</li>
		<li>在每个进料室内包括挡板;挡板是一个 PVC 件, 是3厘米的高度和6厘米宽, 将被放置3.5 厘米从 6 cm 一侧的喂养室, 有孔钻孔15毫米从底部。把挡板粘在房间的底部, 这样水就会流过它。</li>
		<li>包括可移动的第二挡板片, 50 毫米长, t 形件 (58 毫米宽在 t 的底部, 在15毫米从顶部; 它加宽到宽度72毫米)。该形状允许挡板在进料室壁上休息 , 并使水在腔内的挡板动 (图 2c) 。将活动挡板放在双壳的前面 1-2 厘米, 这会迫使水流直接流入房间底部的双壳贝类上。</li>
		<li>将头室和进料装置放在框架表的顶部, 用防滑垫将其放置到位。该系统采用模块化的方式设计, 便于包装、移动和存储。</li>
	</ol>
</li>
</ol><img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58213/58213fig2.jpg"> 图 2: 对头水箱和加料室的详细测量.(一) 这是一幅头部水箱的图纸, 详细测量。(二) 这是一幅有详细量度的食人室图。条纹线指示固定挡板的位置。(三) 这是一幅图画及可移动挡板的量度。<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/58213/58213fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
2. 加料室的流量标定
<ol><li>要校准流速, 请在进料室的出口处放置一个100毫升的玻璃或塑料分级气缸。立即开始用秒表记录时间。</li>
	<li>三十年代后, 取出已毕业的气缸并检查所收集的水的容积。理想情况下, 收集100毫升的水, 这等于从头部水箱到12升-1的喂养室的流量。 注:12 升-1的流速由以前的实验室试验确定, 在不加水循环的情况下, 水族馆之间的颗粒均匀分布。<ol>
<li>如果所收集的水量不在100毫升目标的5毫升以内, 则通过关闭或打开位于头水箱和进料室之间的阀门来调整流量。通过收集三十年代的水再次检查新的流速, 并重复此步骤, 直到获得所需的流速。</li>
	</ol>
</li>
	<li>在数据收集开始之前, 对每个进料室 (包括控制室) 重复相同的校准程序。</li>
</ol>3. Biodeposition 法过滤器的制备
注: 用25毫米直径的 GF/C 玻璃纤维过滤器, 测定水、pseudofeces 和粪便中的总、有机和无机颗粒物。在取样收集之前, 请确保过滤器被洗涤、烘干、烧毁和 preweighed。在所有过程中, 始终使用平尖钳来处理过滤器。如果过滤器断开或有孔, 则丢弃它而不使用它。
<ol><li>要清洗过滤器, 首先, 增加大约10过滤器到一个烧杯与200毫升蒸馏水并且手工搅动他们。在十五年代以后, 注意以前清澈的水有白色纤维在它;这些是松散的灰尘般的玻璃纤维释放的过滤器。别搅了</li>
	<li>醒酒烧杯中的水, 再加200毫升蒸馏水。彻底清洗过滤器3x。重复洗涤过程, 直到有足够的过滤器进行充分的喂养实验, 即约48过滤器的水过滤如果实验持续2小时, 水收集每15分钟, 和32过滤器的粪便和 pseudofeces 的16双壳.</li>
	<li>将过滤器干燥60摄氏度, 至少1小时, 在450摄氏度的消音器炉中烧干过滤器, 以去除任何污染有机物。从熔炉中取出过滤器, 将它们转移到干燥, 并允许过滤器进入室温。</li>
	<li>权衡分析天平上的过滤器, 并记录重量。跟踪过滤器权重的两种可能的方法如下所示。<ol>
<li>在非常边缘的每个过滤器编号, 在过滤过程中接收样品的区域之外, 使用软铅笔。在对过滤器进行编号后对其进行称量, 在笔记本中记录其数量和重量, 并在其原始过滤器框中称量后存储过滤器。</li>
		<li>单独称量每个过滤器, 然后将其包装在一张低沉的铝箔上, 并在箔上记录相应的重量。存储已包装的滤镜, 直到在该字段中使用, 并在收集完样本后记下笔记本的重量。</li>
	</ol>
</li>
</ol>4. 肠道转运时间
<ol><li>将五双壳贝类单独放置在玻璃或塑料烧杯中, 填充300毫升的环境, 未经过滤的海水。</li>
	<li>在每个烧杯中加入2毫升的卡德藻, 记录每只双壳贝类打开的时间, 这是由一个贝壳发出的哈欠发出的信号。 注:卡德藻sp 用于确定肠道转运时间, 因为它容易被双壳类所摄取, 由此产生的粪便颜色呈深绿色, 区别于天然的消化后产生的褐色粪便。浮游生物群落。</li>
	<li>每 3-5 分钟检查每个烧杯, 以确保双壳贝类保持开放和生产粪便。<ol>
<li>检查粪便是否密集, 由双壳贝类的消化过程引起的紧密的弦 (图 3), 并保持其结构时 pipetted。</li>
		<li>确保所收集的沉积物为粪便, 而不是 pseudofeces (图 3), 如果生产, 则由于过剩的卡德藻sp 而立即产生;pseudofeces 是轻包装的, 云状的沉积物的非摄取颗粒, 迅速并用重悬时, 收集与吸管。</li>
	</ol>
</li>
</ol><img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58213/58213fig3.jpg"> 图 3: 双壳类粪便与 pseudofeces 之间视觉差异的图示.左面板显示有肋贻贝 (Geukensia demissa), 箭头指示产生的粪便和 pseudofeces。右侧的面板详细地显示了在过滤卡德藻sp 后产生的绿色粪便和 pseudofeces, 以及在过滤了自然浮游植物群落后产生的褐色粪便和 pseudofeces。<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/58213/58213fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="4"><li>当绿色粪便出现时, 记录每双壳的时间。双壳贝类的开放时间长短与绿色粪便的产生是其肠道转运时间。平均所有五种双壳贝类的肠道转运时间, 以获得平均肠道转运时, 用于定时的偏移之间的取样收集水样和粪便样本。 注: 使用五复制, 以防一个或多个贝类无法打开或产生粪便。理想情况下, 平均肠道转运时间将基于三多个复制。</li>
</ol>5. 样品收集
<ol><li>收集从头水箱溢出的水样本, 控制室的水, 其中含有在实验中使用的同一双壳类的空壳 (每侧两个), 以及每双壳生产的粪便和 pseudofeces。清洁 epibionts 和其他不惑之年鲈生物的双壳贝类, 以避免其他动物在将双壳贝类放入喂食室之前进行过滤。 注: 放置在喂食室中的双壳贝类可以走动, 以便方便粪便和 pseudofeces 收集, 固定在每个房间内使用紧固件 (e. g,尼龙搭扣)。<ol>
<li>每15分钟收集300毫升水2小时, 分别通过 preweighed 过滤器 (即每时间点3个过滤器) 将溢水和水从两组控制室中过滤出来。当过滤器仍在过滤歧管上时, 用5毫升的等渗甲酸铵冲洗过滤器。</li>
		<li>将 biodeposit 收集的起始时间从收集的水中延迟到根据议定书4节所述确定的平均肠道运输时限的长度。例如, 如果平均肠道转运时间为1小时, 当喂食室中的双壳贝类打开时, 立即开始收集水。1小时后, 清除所有已生产的粪便和 pseudofeces 的分庭, 然后开始收集所有随后的粪便和 pseudofeces。<ol>
<li>在喂食室和肠道中转容器中遮蔽双壳贝类, 以增加开放饲料的贝类数量。</li>
		</ol>
</li>
		<li>收集粪便和 pseudofeces 与玻璃吸管, 并保持 biodeposits 在一个单独的容器 (烧瓶或管) 为每双壳在整个2小时收集期间。将每个容器中的 biodeposits 分别过滤到 preweighed 过滤器上, 并用5毫升的等位甲酸铵冲洗。 注: 在2小时的收集结束时, 将有16个集装箱与粪便收集和16个集装箱与 pseudofeces 收集, 总共32个容器过滤。</li>
		<li>将过滤器存储在培养皿中或在闷着的铝箔上, 用于输送到实验室。如果用低沉的铝箔用于运输, 首先将过滤器折成一半, 用过滤材料放在褶皱内部, 以防止通过与箔接触而导致过滤材料的任何损耗。将所有过滤器存储在冷却器和冰块中。</li>
		<li>在实验室, 烘干所有过滤器在烤箱在60°c 至少24小时。</li>
		<li>使用分析平衡称量含机油每个过滤器。减去从最终重量的初始重量, 以确定总颗粒物质。</li>
		<li>将450摄氏度的消音器炉内所有过滤器烧掉4小时. 从熔炉中取出过滤器, 将其转移到干燥, 并允许过滤器进入室温。在分析天平上重新称量过滤器。从干式过滤器重量中减去被烧毁的过滤器重量, 以确定微粒无机物质。 注: 颗粒有机质是颗粒物与微粒无机物质的区别。</li>
	</ol>
</li>
</ol>

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P., Vazquez, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modelling+of+interactions+between+inshore+and+offshore+aquaculture.">Modelling of interactions between inshore and offshore aquaculture.</a> Aquaculture. 426, 154-164 (2014).</li><li>Stevens, C., Plew, D., Hartstein, N., Fredriksson, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+physics+of+open-water+shellfish+aquaculture.">The physics of open-water shellfish aquaculture.</a> Aquacultural Engineering. 38 (3), 145-160 (2008).</li><li>Iglesias, J. I. P., Urrutia, M. 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U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficiency+of+particle+retention+in+13+species+of+suspension+feeding+bivalves.">Efficiency of particle retention in 13 species of suspension feeding bivalves.</a> Ophelia. 17, 239-246 (1978).</li><li>Shumway, S. E., Cucci, T. L., Newell, R. C., Yentsch, 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=Particle+selection,+ingestion,+and+absorption+in+filter-feeding+bivalves.">Particle selection, ingestion, and absorption in filter-feeding bivalves.</a> Journal of Experimental Marine Biology and Ecology. 91 (1-2), 77-92 (1985).</li><li>Velasco, L. A., Navarro, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeding+physiology+of+two+bivalves+under+laboratory+and+field+conditions+in+response+to+variable+food+concentrations.">Feeding physiology of two bivalves under laboratory and field conditions in response to variable food concentrations.</a> Marine Ecology Progress Series. 291, 115-124 (2005).</li><li>Galimany, E., Ram&#243;n, M., Ibarrola, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeding+behavior+of+the+mussel+Mytilus+galloprovincialis+(L.)+in+a+Mediterranean+estuary:+A+field+study.">Feeding behavior of the mussel Mytilus galloprovincialis (L.) in a Mediterranean estuary: A field study.</a> Aquaculture. 314 (1-4), 236-243 (2011).</li><li>Wilcox, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Understanding and applying basic statistical methods using R. , (2017).</li><li>Velasco, L. A., Navarro, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeding+physiology+of+two+bivalves+under+laboratory+and+field+conditions+in+response+to+variable+food+concentrations.">Feeding physiology of two bivalves under laboratory and field conditions in response to variable food concentrations.</a> Marine Ecology Progress Series. 291, 115-124 (2005).</li><li>Filgueira, R., Labarta, U., Fern&#225;ndez-Reiriz, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow-through+chamber+method+for+clearance+rate+measurements+in+bivalves:+design+and+validation+of+individual+chambers+and+mesocosm.">Flow-through chamber method for clearance rate measurements in bivalves: design and validation of individual chambers and mesocosm.</a> Limnology and Oceanography Methods. 4, 284-292 (2006).</li><li>Grizzle, R. E., Greene, J. K., Luckenbach, M. W., Coen, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+in+situ+method+for+measuring+seston+uptake+by+suspension-feeding+bivalve+molluscs.">A new in situ method for measuring seston uptake by suspension-feeding bivalve molluscs.</a> Journal of Shellfish Research. 25 (2), 643-649 (2006).</li><li>Riisg&#229;rd, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+measurement+of+filtration+rates+in+bivalves+-+the+stony+road+to+reliable+data:+review+and+interpretation.">On measurement of filtration rates in bivalves - the stony road to reliable data: review and interpretation.</a> Marine Ecology Progress Series. 211, 275-291 (2001).</li><li>Newell, C. R., Wildish, D. J., MacDonald, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+velocity+and+seston+concentration+on+the+exhalant+siphon+area,+valve+gape+and+filtration+rate+of+the+mussel+Mytilus+edulis.">The effects of velocity and seston concentration on the exhalant siphon area, valve gape and filtration rate of the mussel Mytilus edulis.</a> Journal of Experimental Marine Biology and Ecology. 262 (1), 91-111 (2001).</li></ol>

作者没有什么可透露的。

采用不同的方法对双壳贝类在实验室和田间的过滤和饲喂进行了研究。在使用天然 seston 时所做的测量将会产生最类似于自然环境24的喂养率。现有的用于测量双壳饲养25、26的便携喂养装置依赖于稳定的平台, 如土地或固定船坞;因此, 在这一领域, 对双壳贝类的过滤和喂养进行量化, 直到目前为止, 仅限于非常近岸水域。这里提出的新的仪器和方法代表了一个可靠的工具, 以量化在近海海域的双壳贝类的喂养性能, 在这种情况下, 贝类与环境之间的相互作用已被描述得很差。
biodeposition 方法在近海应用中的关键步骤包括: (1) 头水箱的曝气和所有进料室的流速校准, 以确保均匀的颗粒分布到双壳贝类;(2) 在 biodeposits 收集前, 准确确定实验性肠道转运时间;(3) 对双壳贝类生产的所有粪便和 pseudofeces 进行鉴定、分离和完整收集, 包括收集足够的 biodeposits 以超过有机和无机颗粒物的检测极限。由于 refiltration18、25、27、28等因素, 高流速对避免进料室的水循环有重要的意义, 可能会增加食品浓度减少现象。
在近海环境中, 粪便和 pseudofeces 的准确识别和分离是很有挑战性的。马萨诸塞州水域的粪便和 pseudofeces 的收集在测量的最后一个小时内很可能受到大浪的影响。使用这种方法的测量将受到海洋状态的限制, 影响采摘者清洁地分离和准确区分粪便、pseudofeces 和其他颗粒物 (如淤泥或颗粒) 的能力。喂食室。这一实验问题可以在结果数据中观察到, 在那里, pseudofeces 的有机含量比其他两个位置有更大的变异性 (图 7)。
有非常低颗粒物质的地点, 例如加利福尼亚, 将提出一个分析的挑战, 因为在这个实验中收集的微粒物质非常接近检测的极限, 即使每水样品过滤了 2 L 水。对总颗粒物的有机和无机贡献进行量化的方法是以质量平衡为基础的;因此, 在检测极限附近的小分析误差可能导致生理上不可能的贝类喂养结果, 如负排斥或清除率。此类错误的数据和适当的校正, 如图 8所示, 它绘制了清除率、过滤速率和加利福尼亚实验的吸收率的平均值。粪便数量是如此之小, 在这个地方, 一些被误认为 pseudofeces 由 biodeposit 采摘。收集的极少量 "pseudofeces" 与重量检测的极限非常接近, 由此产生的数据为几种复制提供了阴性贝类过滤和喂养数据, 这在生理上是不可能的, 因此,显然不正确。接近检测极限的微粒物质也产生了很高的变异性。这些结果可能是由于对过滤器称重的错误造成的, 但更有可能的原因是 pseudofeces 的识别不正确。后者的可能性得到进一步支持的观察, 即水总颗粒物太低, 以触发 pseudofeces 生产22,23。通过丢弃不正确的 pseudofeces 数据并只计算摄取通路来纠正数据 (图 8)。
在船上使用 biodeposition 法定量双壳贝类悬浮饲料的装置可以修改和适应几种双壳类。喂食室的大小可以略有变化, 以适应更宽或更窄的双壳壳。然而, 重要的是要注意, 改变喂食室的尺寸从这里所描述的, 要求在进行任何测量之前建立在喂食室的均匀颗粒分布。滤水量应根据当地条件进行调整。低 seston 的环境, 如加利福尼亚, 需要大量的水过滤超过检测限额的重量为基础的分析。同时, 如果过滤太多的水, 过滤器堵塞, 烤箱的干燥时间 (不温度) 需要增加。同样, biodeposit 集合可能需要在低 seston 环境中加长, 以收集足够的材料以超过分析检测限制。另一个有问题的 biodeposit 收集的指标是水与 pseudofeces 和粪便的相对有机含量。粪便和 pseudofeces 不得含有比水更大的有机物百分比;它们是从水中过滤和处理过的微粒的产物。在某些条件下, biodeposits 的有机含量可能略大于水, 因为双壳制加工食品颗粒的有机投资;然而, 这项投资至多会对粪便有机物产生轻微的增加。在这里报告的有机物的百分比远远高于可能归因于代谢性粪便损失的百分比。来自马萨诸塞州的 pseudofeces 样本说明了这个潜在的问题。如上所述, pseudofeces 的有机含量相当多变, 但有些复制产生的有机物含量大大超过了相应的水样。在 biodeposit 收集的最后一个小时的海洋中, pseudofeces 与外源有机物相结合, 人为地提升有机含量, 产生生理上不可能的结果 (图 7).如果公海国家在今后应用这种方法时有可能出现, 那么建议通过增加分庭增加更多的复制。
该方法的局限性在于该仪器旨在量化成人个体的喂养。由于 (伪) 粪便的体积小, 需要较长的实验以获得足够的材料, 以达到分析检测的极限, 因此, 从双壳种子中准确、完整地收集粪便和 pseudofeces 是很困难的。如果使用的是小型个体, 则可以在一个房间里汇集几个人, 以增加每室的粪便和 pseudofeces 产量。另外, 可以用更小的实验室重新设计这些设备。天气和海洋状态也可能是重要的限制, 因为这将影响 biodeposit 样品收集的准确性。极端温度和降雨可能会减少饲料中双壳的复制数量。在实验中, 对水泵的部署深度可能会有所不同, 以确保实验中使用的 seston 反映出双壳类养殖的深度 seston 典型。尽管有这些潜在的限制, 该方法提供了独特的机会, 以研究在自然条件下的贝类的过滤和喂养, 与自然 seston, 而不是模拟条件在实验室。所产生的数据比实验室实验更切合实际, 更有可能反映出在感兴趣地点的双壳贝类的性能。新的舰船测量方法大大扩大了潜在的地理范围。
对近海贻贝养殖的兴趣日益浓厚, 为今后应用此方法提供了一个理想的用户群。有意优化近海水产养殖业务选址的利益相关者可以利用这种方法检查拟议地点的双壳贝类性能。正在计划中的应用程序的一个例子是测试新英格兰南部沿岸海域的蓝贻贝悬浮培养最佳深度的假说 (Mizuta 和 Wikfors)。

野生捕捞渔业在全球范围内下降了1。因此, 未来海产品供应的增长必须来自水产养殖的扩大。水产养殖的生产一直在增长, 并将继续快速增长, 通过 2025, 使水生养殖最迅速增长的粮食生产系统2。养殖悬浮喂养的双壳贝类 (贻贝、牡蛎、扇贝和蛤) 被认为是最环保的水产养殖形式之一, 因为这些有机体不需要额外的喂养, 而是获得营养从自然浮游植物生产和转移有机质到底栖有机体3,4。事实上, 贝类养殖被认为是改善富营养化河口5、6的水质和营养结构的合法工具。尽管沿海海湾和河口的贝类养殖发展前景普遍看好, 但与其他沿海海洋利益的冲突, 如商业和娱乐渔业、娱乐活动和美学沿海土地所有者的愿望--在 "社会承载能力" 一词下聚集的社会限制, 使一些人看到了 "开阔的海洋", 以便大规模扩大贝类养殖7。
在公海上移动贝类养殖, 为贝类养殖扩展提供了巨大的潜力, 但也给海洋生态系统8的生物体带来了前所未有的挑战。首先, 大多数养殖的、悬浮喂养的双壳贝类是河口生物体, 它们在不同的环境中进化, 从9的开放海洋生态系统中有许多差异。盐度、温度和水化学的季节性和昼夜时间变化, 以及沿海水域高和可变养分供应所刺激的强烈生物活性选择了行为和生理贻贝、牡蛎、扇贝和蛤的特性, 在相对恒定的、稀释的海洋环境中, 可能没有什么好处10。众所周知, 双壳贝类应对这些环境变化作出反应, 调节其过滤, 利用良好的水质周期, 并优化其食品购置11,12。在一个更加恒定的环境中, 如开阔水域, 不清楚双壳贝类是否能有效地调节其泵浦和过滤速率, 以维持一个快速增长的正能量平衡。近海贝类养殖面临的第二个挑战也与海洋中相对较低的 seston 食物供应有关。由于浮游植物密度远低于河口, 目前在河口成功养殖的双壳贝类是否能找到足够的食物来维持新陈代谢和生长？目前使用线、袜子、笼子或其他围栏在河口饲养贝类的做法导致三维过滤器, 即使在富营养化的沿海水域13,14, 也可能在当地消耗浮游植物。对养殖齿轮的设计、放养密度、线间距、作物周期时间等方面的假设需要重新考虑, 以管理农场的生产承载能力和当地海洋生态系统的生态承载能力。15,16. 在近岸进行的集约化贝类养殖可能需要加以修改, 以适应海洋的稀薄环境。
为了进一步了解沿海贝类养殖的做法可能需要加以修改, 以便在近海取得成功, 关于贝类与近海地区的 seston 之间如何相互作用的定量数据是必不可少的。通过悬浮喂养的双壳贝类对微粒进行定量过滤、清除、摄取、排斥和吸收的技术已经发展了17、18。其中一些方法已被优化, 以检测在非常短的时间尺度上的变化, 不同的粒子类型之间的选择, 或对各种环境变化的生理反应19,20,21.最近, 改进所谓的 biodeposition 方法已导致接受这种方法作为一个合法的工具, 以量化大多数重要的过滤和喂养变量的贻贝, 牡蛎, 蛤17,22.
biodeposition 方法, 一般, 使用一个质量平衡的方法, 以无机 seston 组分作为追踪者, 定量划分由各自的贝类有机和无机 seston 组分成比例被捕获, 被拒绝, 被摄取,并吸收超过17小时的时间刻度。为使这种方法准确, 至关重要的是, 运送到个别贝类的水流速率是恒定的, 确切地说是已知的, 而且贝类不会受到身体的干扰, 从而保持其不断的过滤行为。此外, 还必须同步收集在双壳贝类进食时的水样, 并收集消化后产生的粪便样本 (即排遗)。这两个过程 (摄取和排遗) 是抵消了一个微粒物质通过双壳肠道转运所需的时间长短。肠道转运时间表示摄取食物和以粪便形式释放未消化物质之间的时间。此外, 从实际的角度来看, biodeposits 需要由研究人员定量收集, 然后再按水运动分类。由于这些原因, 使用 biodeposition 方法对双壳类过滤进行量化的器具和程序仅限于非常近岸的地点, 那里有稳定的平台--干燥的土地或固定的码头--足够接近贝类种群研究。为了在近海使用 biodeposition 方法, 有必要找到一种满足船上稳定平台的方法要求的方法。
几个世纪前, 水手寻求解决相同的基本问题, 如何隔离船上的文章从船舶的运动发展了框架。一个框架在连接到船的平台和被隔离的文章之间引入一个或多个支点, 使孤立的文章对引力的反应比对船的运动更有响应。我们使用了也许最简单的框架设计-别针枢轴在90°角度-在设计一个设备修改了从由 Galimany 和工友报告的一个22。在本报告中, 该仪器的有效功能通过测量验证: 1) 与船的运动相比, 表与贝类室的运动, 2) 流速通过20个复制分庭在海上, 3) 的一致性在三艘不同船只上三个离岸地点测试的贻贝过滤数据。

<table>
	<tbody>
		<tr>
			<td>GF/C glass microfibre filters</td>
			<td>Whatman</td>
			<td>1822-025</td>
			<td>25 mm diameter circles</td>
		</tr>
		<tr>
			<td>Submersible Utility Pump</td>
			<td>Utilitech</td>
			<td>PPSU33</td>
			<td>1/3 HP</td>
		</tr>
		<tr>
			<td>Filtration manifold</td>
			<td>Sterlitech</td>
			<td>313400</td>
			<td>3-place manifold, PVC</td>
		</tr>
		<tr>
			<td>Filter forceps</td>
			<td>Millipore</td>
			<td>XX6200006P</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter funnel</td>
			<td>Ace Glass</td>
			<td>D140942</td>
			<td>300 ml; glass</td>
		</tr>
		<tr>
			<td>Frit support</td>
			<td>Fisher Scientific</td>
			<td>09-753-14</td>
			<td>25mm diameter; glass</td>
		</tr>
		<tr>
			<td>Vacuum Filter Holders</td>
			<td>Fisher Scientific</td>
			<td>09-753-4</td>
			<td>For 25mm filter funnels and frit supports</td>
		</tr>
		<tr>
			<td>Drying Oven</td>
			<td>Fisher Scientific</td>
			<td>15-103-0503</td>
			<td>Gravity convection</td>
		</tr>
		<tr>
			<td>Box Furnace Oven</td>
			<td>ThermoFisher Scientific</td>
			<td>BF51794C</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium formate</td>
			<td>Fisher Scientific</td>
			<td>A666-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetraselmis sp.</td>
			<td>National Center for Marine Algae and Microbiota</td>
			<td></td>
			<td>119 strains of Tetraselmis sp. are available for sale by NCMA, and specific strain should be selected based on temperature of planned experiments. As such, we have not recommended a specific catalog number here.</td>
		</tr>
		<tr>
			<td>Glass petri dish</td>
			<td>Fisher Scientific</td>
			<td>08-747A</td>
			<td>60 mm diameter</td>
		</tr>
	</tbody>
</table>

design and use of an apparatus for quantifying bivalve suspension feeding at sea

由于贝类养殖从沿海海湾和河口转移到离岸地区, 需要量化养殖的双壳贝类 (即贻贝、牡蛎和蛤) 的生态系统相互作用, 这带来了新的挑战。为了确定离岸贝类养殖场的重要生态系统相互作用, 包括它们的承载能力、与浮游动物群落的竞争, 需要对悬浮式软体动物饲养行为的定量数据,不同深度的营养资源的可用性和海底的沉积。biodeposition 方法用于定量在自然环境下悬浮喂养的双壳贝类中的喂养变量, 代表比实验室实验更现实的代理。然而, 这种方法依赖于一个稳定的平台, 以满足对贝类提供的水流速率保持不变和双壳贝类不受干扰的要求。通过在该装置周围建立一个二维框架表, 通过使用 biodeposition 方法对双壳贝类的饲养量进行量化的流动装置和过程。Planimeter 数据揭示了包含试验贝类的腔室的最小间距和偏航尽管船运动, 房间内的流速保持不变, 操作员能够收集 biodeposits (粪便和 pseudofeces), 足够一致性, 以获得准确测量的双壳贝类的清除, 过滤, 选择, 摄取, 排斥, 并吸收在近海贝壳养殖地点。

biodeposition 方法确定了双壳饲养的数量, 并提供了一种利用自然 seston 在野外环境中获取双壳贝类过滤和饲喂性能的综合数据的机制。biodeposition 方法以前的应用只能在岸上的位置进行, 因为该方法需要一个稳定的平台。对近海海域的双壳贝类过滤和饲养的研究需要以船舶为基础的测量, 而船舶在平淡条件下也不够稳定。我们已经设计和测试了添加一个框架表到现有的滤料装置, 以创建一个稳定的平台, 需要正确使用 biodeposition 方法。
随着双壳贝类的稳定平台的过滤, 我们报告的数据表明, 均匀颗粒分布在每个房间内的喂食装置 (p = 0.997 从一个泛化的韦尔奇的测试20% 修剪手段23;图 4)。这种均匀分布的悬浮物表明, 将颗粒从头水箱输送到单个腔体是一致的;因此, 所有的双壳贝类都暴露在相同的食物数量和质量, 可以被认为是真实的复制。
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58213/58213fig4.jpg"> 图 4: 空腔颗粒分布试验中每个加料室的平均细胞丰度.此面板显示了在质量保证试验期间从每个喂食室的出口管收集的海水中浮游植物细胞/mL (@ SD) 的平均数量 (标记为 1-20), 以确保颗粒在流经系统中均匀分布。<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/58213/58213fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
在三个地点进行了四次船上试验, 其中三种贻贝品种的 seston 量和成分非常不同 (图 5)。研究的不同物种可能是或目前正在近海养殖;我们用多个物种来测试仪器的一般适用性。蓝贻贝 (贻) 被用于第一个康涅狄格州 (CT) 实验和马萨诸塞州 (MA)。第二次 CT 实验采用肋贻贝 (Geukensia demissa)。地中海贻贝 (贻地中海) 在加利福尼亚 (CA) 实验中使用。两个实验是在海岸 CT, 在长岛的声音, 1.5 公里的米尔福米在2013年6月12日和2013年6月19日。第三个实验是在1公里外的 Menemsha 在 2013年7月23日, 在葡萄园的声音, 在沿海马进行。第四项试验是在2013年8月20日离长滩10公里的近海 CA 进行的。
在这三个地点的条件范围涵盖了对贝类水产养殖进行评估的近海环境的预期。水总颗粒物的 CT 最高, MA 低, CA 最低 (所有p≤0.001 从 Dunnett 的 T3 程序的推广和引导t技术23)。与此相反, seston 的有机含量最高, MA 低, CT 最低 (所有p≤ 0.01, 从 Dunnett 的 T3 程序的推广到修剪方法和引导t技术23;图 5)。
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/58213/58213fig5.jpg"> 图 5: 三个实验地点水中颗粒物的组成和数量.该面板显示了平均颗粒有机质 (POM) 和灰色的数据和误差条, 以及平均微粒无机物质 (PIM), 从3个不同实验地点收集到的水中的白色和误差条中的数据。整条 (灰 + 白) 表示颗粒物总量 (TPM)。CT 1 = 康涅狄格实验 1;CT 2 = 康涅狄格实验 2;马萨诸塞州实验;加利福尼亚州实验。<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/58213/58213fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
贝类饲养行为既依赖于物种, 又依赖于环境条件。个体根据水中微粒物质的数量和类型 (有机和无机) 的差异来调整喂养行为。因此, 来自三个地点的四种滤食性实验的结果反映了对食物数量和质量的塑性生理反应, 以及四实验中三的物种差异。在第一次 ct 实验中, 贻贝的吸收效率明显高于第二次, 而第一次 ct 实验比 CA 高, 但所有配对比较均无显著性, 可能是由于在MA 和 CA 测量 (在α = 0.05 测试的意义, 调整到控制多项测试; 从 Dunnett 的 T3 程序的推广到修剪方法和引导t技术;23图 6)。被拒绝的被过滤的材料的比例是最高的 CT, 低在 MA, 并且是零在 CA (所有p≤0.005 从 Dunnett 的 T3 做法的概括为修剪的手段和引导t技术23)。
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/58213/58213fig6.jpg"> 图 6: 在船上试验中, 贻贝对总颗粒物的排斥和有机物的吸收.该面板显示了贻贝在三个实验地点的排斥和吸收百分比。CT 1 = 康涅狄格实验 1;CT 2 = 康涅狄格实验 2;马萨诸塞州实验;加利福尼亚州实验。蓝贻贝 (贻) 用于 CT 1 和 MA。肋贻贝 (Geukensia demissa) 用于 CT 2。地中海贻贝 (贻地中海) 在加利福尼亚州使用.<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/58213/58213fig6large.jpg" target="_blank">请点击这里查看这个数字的大版本.</a>
MA 和 CA 的实验说明了在环境条件变化过程中可能出现的常见问题。高海洋状态导致 MA 中 pseudofeces 的测定有机含量有较高的相对变异性。
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/58213/58213fig7v2.jpg"> 图 7: 三个实验地点的水、粪便和 pseudofeces 的有机含量.这一小组显示了在3个地点进行的四项不同试验中, 三种贻贝种类的水和粪便中有机质 (pseudofeces) 的平均百分比。CT 1 = 康涅狄格实验1与蓝色贻贝 (贻);CT 2 = 康涅狄格实验2与肋贻贝 (Geukensia demissa);马萨诸塞州实验用蓝色贻贝;加利福尼亚州的实验与地中海贻贝 (贻地中海)。<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/58213/58213fig7v2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
从 CA 的喂养行为结果中, 说明了通常与低颗粒物质区相关的分析问题, 其中一些小 pseudofeces 最初被误认为是粪便。
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/58213/58213fig8.jpg"> 图 8: biodeposits 的误认对船载试验中贻贝的摄食行为数据的影响.该面板显示了来自加利福尼亚的样本数据, 显示了 misidentifying 小粪便在低总颗粒物质 (TPM) 环境中的 pseudofeces 效应。在这种情况下, TPM 太低, 无法触发 pseudofeces 生产, 但粪便太小, 有些被误认为是 pseudofeces。数据通过结合粪便和 "pseudofeces" 的重量和只计算摄取途径来纠正。CR = 清除率, 通过贻贝 (L/小时) 鳃循环的水量;FR = 过滤速率, 残留在鳃中的颗粒量 (毫克/小时);AR = 吸收率, 吸收在贻贝消化系统 (毫克/小时) 摄取的微粒物质的数量。<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/58213/58213fig8large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
如图 7和图 8所示的案例研究将在讨论部分更详细地说明。

Watch this Scientific Journal Video about Design and Use of an Apparatus for Quantifying Bivalve Suspension Feeding at Sea at JoVE.com

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

海上双壳贝类悬浮饲料定量装置的设计与应用

采用 biodeposition 方法对双壳贝类的过滤和饲养行为进行了定量的流动装置的改进, 供船用。在该装置周围建立的二维框架表将设备与船的运动隔离开来, 从而使近海贝类养殖地的双壳类过滤变量得到准确的量化。

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

design-use-an-apparatus-for-quantifying-bivalve-suspension-feeding-at

Universidad de Cartagena UDEC

Research

JoVE Journal

Environment

7.7K 次观看.  National Oceanic and Atmospheric Administration (NOAA) Fisheries Service. 采用 biodeposition 方法对双壳贝类的过滤和饲养行为进行了定量的流动装置的改进, 供船用。在该装置周围建立的二维框架表将设备与船的运动隔离开来, 从而使近海贝类养殖地的双壳类过滤变量得到准确的量化。

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

Design and Construction of an Urban Runoff Research Facility

As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use due to increased demand for landscape irrigation. Improper irrigation practices and large rainfall events can result in runoff from urban landscapes which has potential to carry nutrients and sediments into local streams and lakes where they may contribute to eutrophication. A 1,000 m2 facility was constructed which consists of 24 individual 33.6 m2 field plots, each equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated urban landscapes. Runoff volumes from the first and second trials had coefficient of variability (CV) values of 38.2 and 28.7%, respectively. CV values for runoff pH, EC, and Na concentration for both trials were all under 10%. Concentrations of DOC, TDN, DON, PO4-P, K+, Mg2+, and Ca2+ had CV values less than 50% in both trials. Overall, the results of testing performed after sod installation at the facility indicated good uniformity between plots for runoff volumes and chemical constituents. The large plot size is sufficient to include much of the natural variability and therefore provides better simulation of urban landscape ecosystems.

This paper describes the design, construction, and function of a 1,000 m2 facility containing 24 individual 33.6 m2 field plots equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated home lawns.

设计一个城市径流研究设施的建设

As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use ...

科研

环境科学

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 ...

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 ...

Assaying Predatory Feeding Behaviors in Pristionchus and Other Nematodes

This protocol provides multiple methods for the analysis and quantification of predatory feeding behaviors in nematodes. Many nematode species including Pristionchus pacificus display complex behaviors, the most striking of which is the predation of other nematode larvae. However, as these behaviors are absent in the model organism Caenorhabditis elegans, they have thus far only recently been described in detail along with the development of reliable behavioral assays 1. These predatory behaviors are dependent upon phenotypically plastic but fixed mouth morphs making the correct identification and categorization of these animals essential. In P. pacificus there are two mouth types, the stenostomatous and eurystomatous morphs 2, with only the wide mouthed eurystomatous containing an extra tooth and being capable of killing other nematode larvae. Through the isolation of an abundance of size matched prey larvae and subsequent exposure to predatory nematodes, assays including both "corpse assays" and "bite assays" on correctly identified mouth morph nematodes are possible. These assays provide a means to rapidly quantify predation success rates and provide a detailed behavioral analysis of individual nematodes engaged in predatory feeding activities. In addition, with the use of a high-speed camera, visualization of changes in pharyngeal activity including tooth and pumping dynamics are also possible.

Assaying Predatory Feeding Behaviors in Pristionchus and Other Nematodes

Behavioral assays provide powerful tools for understanding neuronal function. Here we present several protocols for quantifying predatory feeding behavior found in the model nematode Pristionchus pacificus and its relatives. Additionally, we provide methods for analyzing predatory feeding adaptations including mouth structures and teeth.

测定掠夺取食行为<em&gt; Pristionchus</em&gt;及其他线虫

This protocol provides multiple methods for the analysis and quantification of predatory feeding behaviors in nematodes. Many nematode species including ...

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 ...

Protocols for Quantifying Transferable Pesticide Residues in Turfgrass Systems

Plant canopies in established turfgrass systems can intercept an appreciable amount of sprayed pesticides, which can be transferred through various routes onto humans. For this reason, transferable pesticide residue experiments are required for registration and re-registration by the United States Environmental Protection Agency (USEPA). Although such experiments are required, limited specificity is required pertaining to experimental approach. Experimental approaches used to assess pesticide transfer to humans including hand wiping with cotton gloves, modified California roller (moving a roller of known mass over cotton cloth) and soccer ball roll (ball wrapped with sorbent strip) over three treated turfgrass species (creeping bentgrass, hybrid bermudagrass and tall fescue maintained at 0.4, 5 and 9 cm, respectively) are presented. The modified California roller is the most extensively utilized approach to date, and is best suited for use at low mowing heights due to its reproducibility and large sampling area. The soccer ball roll is a less aggressive transfer approach; however, it mimics a very common occurrence in the most popular international sport, and has many implications for nondietary pesticide exposure from hand-to-mouth contact. Additionally, this approach may be adjusted for other athletic activities with limited modification. Hand wiping is the best approach to transfer pesticides at higher mowing heights, as roller-based approaches can lay blades over; however, it is more subjective due to more variable sampling pressure. Utility of these methods across turfgrass species is presented, and additional considerations to conduct transferable pesticide residue research in turfgrass systems are discussed.

Transferable pesticide residue experiments in turfgrass systems are integral components of human risk exposure assessments. Experimental approaches to measure transferable residues should be adjusted to the human interaction of interest and turfgrass system dynamics. Three transferable pesticide residue protocols are presented and the suitability across three turfgrass systems is discussed.

协议在草坪系统量化转换农药残留

Plant canopies in established turfgrass systems can intercept an appreciable amount of sprayed pesticides, which can be transferred through various routes ...

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.

海洋浮游生物和悬浮颗粒的超洁净多层采集装置

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

The Calibration and Use of Capacitance Sensors to Monitor Stem Water Content in Trees

Water transport and storage through the soil-plant-atmosphere continuum is critical to the terrestrial water cycle, and has become a major research focus area. Biomass capacitance plays an integral role in the avoidance of hydraulic impairment to transpiration. However, high temporal resolution measurements of dynamic changes in the hydraulic capacitance of large trees are rare. Here, we present procedures for the calibration and use of capacitance sensors, typically used to monitor soil water content, to measure the volumetric water content in trees in the field. Frequency domain reflectometry-style observations are sensitive to the density of the media being studied. Therefore, it is necessary to perform species-specific calibrations to convert from the sensor-reported values of dielectric permittivity to volumetric water content. Calibration is performed on a harvested branch or stem cut into segments that are dried or re-hydrated to produce a full range of water contents used to generate a best-fit regression with sensor observations. Sensors are inserted into calibration segments or installed in trees after pre-drilling holes to a tolerance fit using a fabricated template to ensure proper drill alignment. Special care is taken to ensure that sensor tines make good contact with the surrounding media, while allowing them to be inserted without excessive force. Volumetric water content dynamics observed via the presented methodology align with sap flow measurements recorded using thermal dissipation techniques and environmental forcing data. Biomass water content data can be used to observe the onset of water stress, drought response and recovery, and has the potential to be applied to the calibration and evaluation of new plant-level hydrodynamics models, as well as to the partitioning of remotely sensed moisture products into above- and belowground components.

The hydraulic capacitance of biomass is a key component of the vegetation water budget, which serves as a buffer against short and long-term drought stresses. Here, we present a protocol for the calibration and use of soil moisture capacitance sensors to monitor water content in the stems of large trees.

电容式传感器监测林木茎含水量的标定及应用

Water transport and storage through the soil-plant-atmosphere continuum is critical to the terrestrial water cycle, and has become a major research focus ...

Data Collection on Marine Litter Ingestion in Sea Turtles and Thresholds for Good Environmental Status

The following protocol is intended to respond to the requirements set by the European Union&#39;s Marine Strategy Framework Directives (MSFD) for the D10C3 Criteria reported in the Commission Decision (EU), related to the amount of litter ingested by marine animals. Standardized methodologies for extracting litter items ingested from dead sea turtles along with guidelines on data analysis are provided. The protocol starts with the collection of dead sea turtles and classification of samples according to the decomposition status. Turtle necropsy must be performed in authorized centers and the protocol described here explains the best procedure for gastrointestinal (GI) tract isolation. The three parts of the GI (esophagus, stomach, intestine) should be separated, opened lengthways and contents filtered using a 1 mm mesh sieve. The article describes the classification and quantification of ingested litter, classifying GI contents into seven different categories of marine litter and two categories of natural remains. The quantity of ingested litter should be reported as total dry mass (weight in grams, with two decimal places) and abundance (number of items). The protocol proposes two possible scenarios to achieve the Good Environmental Status (GES). First: &#34;There should be less than X% of sea turtles having Y g or more plastic in the GI in samples of 50-100 dead turtles from each sub-region&#34;, where Y is the average weight of plastic ingested and X% is the percentage of sea turtles with more weight (in grams) of plastic than Y. The second one, which considers the food remain versus plastic as a proxy of individual health, is: &#34;There should be less than X% of sea turtles having more weight of plastic (in grams) than food remains in the GI in samples of 50-100 dead turtles from each sub-region&#34;.

The protocol focuses on the collection of sea turtle samples, describing all the steps from the animal recovery and necropsy to the classification and quantification of ingested marine litter. Moreover, the representative results show how to use the collected data to elaborate the possible thresholds for Good Environmental Status.

海洋垃圾摄入海龟和环境状态良好的阈值数据收集

The following protocol is intended to respond to the requirements set by the European Union&#39;s Marine Strategy Framework Directives (MSFD) for the ...

A Method for Quantifying Foliage-Dwelling Arthropods

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density requires a method with high detection probability and a known sampling area. While most described methods provide a qualitative or semi-quantitative estimate adequate for describing species presence, richness, and diversity, few provide an adequately consistent detection probability and known or consistent sampling areas to provide an index or estimate with adequate precision to detect differences in abundance across environmental, spatial, or temporal variables. We describe how to quantify leaf-dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate and quantify them. As we demonstrate, this method can be used at a landscape scale to quantify leaf-dwelling arthropods with adequate precision to test for and describe how spatial, temporal, environmental, and ecological variables influence arthropod richness and abundance. This method allowed us to detect differences in density, richness, and diversity of leaf-dwelling arthropods among 5 genera of trees commonly found in southeastern deciduous forests.

We describe how to quantify leaf dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate for quantification.

定量叶类节肢动物的方法

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density ...

海上双壳贝类悬浮饲料定量装置的设计与应用

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