此协议的发展由IPA亚得里亚海跨境合作计划2007 - 2013年成立后，DeFishGear项目（1°STR / 00010）之内。

1.海面上的塑料微粒采样<ol><li>部署使用用线和登顶三角帆景气或»A型架«船只侧面的蝠鲼网。 </li><li>为了防止收集受尾区域内的湍流水 - （从船上4米距离约3）部署蝠净出唤醒区。 </li><li>写在数据表中的初始GPS坐标和初始时间。 </li><li>开始在一条直线方向约7倍的速度移动。 2 - 3节的30分钟，开始时间测量。 </li><li> 30分钟后停止了船，写下最后的GPS坐标，路线长度（最正确的方法是计算从GPS坐标的长度），平均船速到所提供的数据表和解除曼塔净出水中。 </li><li>从海水中网采用从船上WA潜水泵或水外彻底清洗净曼塔之三水库。冲洗从曼塔嘴鳕鱼端的方向，以集中附着在网成鳕鱼结束所有粒子。 注意:不要通过网络，以防止污染开幕冲洗样本。 </li><li>安全删除鳕鱼端和筛在鳕鱼样品通过一条300微米的筛孔尺寸筛或更少结束。 </li><li>从外部彻底冲洗鳕端，并通过筛倾倒样品的其余部分。直到不再有鳕结束内部的任何颗粒重复此步骤。 </li><li>集中在筛上的所有材料在筛中的一个组成部分。 </li><li>与使用漏斗，冲洗筛入，用70％乙醇的玻璃瓶或塑料瓶。 </li><li>关闭瓶，用纸巾擦拭，并贴上标签的盖子和样品名称和日期防水标志的jar之外（你也应该把对帆纸上用铅笔写了第二个标签在一个罐子里，以避免可能的损失的由于在罐子擦除标签样品名称）。转移标记塑料瓶放入冷藏箱。 注意:一般取样条件:风速不应超过2博福特，因为海浪过高，净是不是海面稳定。在拖网期间保持在恒定速度的平稳线性当然是很重要的。曼塔网开放一半应取样过程中浸没。采样的持续时间应为30分钟（在情况下，有大量的天然材料，如浮游生物，采样的持续时间可以更短）。避免使用塑料工具和容器。避免合成布（如羊毛），绳索和与容器蝠净的接触，以防止样品污染。要非常小心，不要损坏蝠网或船体，同时部署和捕捉网。 </li></ol> 2.分离塑料微粒从海面样本<ol><li>如果样品中不包含大于25毫米的任何产品，似乎是干净的，直接与第3步继续。 </li><li>倾通过筛（≤300微米筛目大小）样品，并删除从样品尺寸&gt; 5毫米（宏和女中音屑）所有的天然或人造的垫料的目的，使用视觉识别和镊子。小心用蒸馏水，以去除附着在任何的微型垃圾仔细冲洗去除每个对象。存放在单独的容器中所有的天然和人造垃圾对象。干燥在干燥器（或在开放的空气，但在一个封闭的培养皿）中的所有天然和人工垫料对象和称重它们。确定根据项目凋落16的类别的主列表中的所有垃圾对象&gt; 25毫米（宏屑）。 </li><li>删除所有较大的物体后，集中所有剩余部分用喷瓶或自来水筛的一部分。用70％的乙醇的最小量与funn的帮助下将样品倒入玻璃容器埃尔。 注:在这一步骤中使用70％的乙醇的关键是要保持样品。此外，在样品的目视检查的步骤，乙醇有助于变色生物体多彩塑料因此变得更容易找到。 </li><li>取样品（子样本）少量，倒入玻璃培养皿。与使用立体显微镜的分析样品（20 - 80倍变焦）并搜索微型塑料颗粒。 </li><li>每个微型塑料粒子应归类到在表1中列出的类别之一，并放入培养皿或其它玻璃小瓶中，标有一个类别名称。培养皿需要在任何时候都被关闭。 注意:当您的样品中分离塑料微粒是保守的，并选择更多，而不是分析更小的颗粒。颗粒的实际化学结构仍将稍后确定。一定要分析来自各方的更大的物体如塑料微粒可能被卡住，因此较大的项目下隐藏。它也可能是有用的已经分析对象移到培养皿的一侧。 </li><li>把培养皿测量设备（由千分尺滑或图像分析软件校准目镜尺），在显微镜下，测量各颗粒的尺寸（测量最长的对角线），除长丝，并记下其颜色。每个子样本应该由其他人进行审查。小心冲洗含有样品，使附着在玻璃墙的所有粒子洗净，放入培养皿玻璃容器。 </li><li>分别通过使用分析级的权衡每个类别的微型塑料颗粒。微型塑料颗粒需要事先称量待干燥。封闭培养皿可以放入一个干燥器或样品可以放置干燥在一个封闭的培养皿直到颗粒变得干燥（封闭培养皿的带颗粒的重量为常数）。 </li><li>确定微型垃圾。在分析搜索塑料微粒的样本，请考虑一些颗粒会轻易可见（颜色，形状，尺寸），而另一些可能是棘手找到。下面是确定样品中的微型颗粒的几个特点:比如，没有细胞结构，参差不齐，尖锐，边缘弯曲，粗细均匀，颜色鲜明（蓝，绿，黄等）。 </li></ol> 3.塑料微粒的化学鉴定<ol><li> ATR-FTIR光谱 <ol><li>在分析之前用酒精清洁无绒布检测系统。 </li><li>记录背景光谱。放置在样品架中样品和收集的光谱。用与数据库中的光谱所得到的频谱的一个自动化比较识别所获得的ATR- FTIR光谱。 </li></ol></li><li> 微ATR-FTIR光谱 <ol><li>在分析之前用酒精清洁无绒布检测系统。 </li><li>放置在玻璃过滤器的样品。注:其他滤镜可我们ED但他们的聚合物性质可以表征干扰。 </li><li>将过滤器与自动扫描台样品，并使用操纵杆来定位样本。 </li><li>记录的光学图像并标记的区域（例如，20 20微米），其中所述样品将被表征。 </li><li>记录背景光谱。 </li><li>放置在样品架中样品，并在预定位置收集的光谱。 </li><li>用与数据库中的光谱所得到的频谱的一个自动化比较识别所获得的微ATR-FTIR光谱。 </li></ol></li></ol>

<ol>
	<li>Law, K. L., 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=Plastic+accumulation+in+the+North+Atlantic+subtropical+gyre.">Plastic accumulation in the North Atlantic subtropical gyre.</a> Science. 329 (5996), 1185-1188 (2010).</li><li>Thompson, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microplastics+in+the+marine+environment:+Sources,+consequences+and+solutions.">Microplastics in the marine environment: Sources, consequences and solutions.</a> Marine anthropogenic litter. , 185-200 (2015).</li><li>Lusher, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microplastics+in+the+marine+environment:+distribution,+interactions+and+effects.">Microplastics in the marine environment: distribution, interactions and effects.</a> Marine anthropogenic litter. , 245-307 (2015).</li><li>Arthur, C., Baker, J., Bamford, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Proceedings of the International Research Workshop on the Occurrence, Effects, and Fate of Microplastic Marine Debris, September 9-11. , (2008).</li><li>Andrady, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microplastics+in+the+marine+environment.">Microplastics in the marine environment.</a> Marine pollution bulletin. 62 (8), 1596-1605 (2011).</li><li>Browne, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sources+and+pathways+of+microplastics+to+habitats.">Sources and pathways of microplastics to habitats.</a> Marine anthropogenic litter. , 229-244 (2015).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marine+litter:+an+analytical+overview.">Marine litter: an analytical overview.</a> UNEP's REGIONAL SEAS PROGRAMME. , (2005).</li><li>van der Wal, M., 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=.">.</a> SFRA0025: Identification and Assessment of Riverine Input of (Marine) Litter, Final Report for the European Commission DG Environment under Framework Contract No ENV.D.2/FRA/2012/0025. , (2015).</li><li>Set&#228;l&#228;, O., Fleming-Lehtinen, V., Lehtiniemi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ingestion+and+transfer+of+microplastics+in+the+planktonic+food+web.">Ingestion and transfer of microplastics in the planktonic food web.</a> Environmental pollution. 185, 77-83 (2014).</li><li>Farrell, P., Nelson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trophic+level+transfer+of+microplastic:+Mytilus+edulis.+(L.)+to+Carcinus+maenas+(L).">Trophic level transfer of microplastic: Mytilus edulis. (L.) to Carcinus maenas (L).</a> Environmental Pollution. 177, 1-3 (2013).</li><li>Wright, S. L., Thompson, R. C., Galloway, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+physical+impacts+of+microplastics+on+marine+organisms:+a+review.">The physical impacts of microplastics on marine organisms: a review.</a> Environmental Pollution. 178, 483-492 (2013).</li><li>Bakir, A., Rowland, S. J., Thompson, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transport+of+persistent+organic+pollutants+by+microplastics+in+estuarine+conditions.">Transport of persistent organic pollutants by microplastics in estuarine conditions.</a> Estuarine, Coastal and Shelf Science. 140, 14-21 (2014).</li><li>Cole, M., Lindeque, P., Halsband, C., Galloway, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microplastics+as+contaminants+in+the+marine+environment:+a+review.">Microplastics as contaminants in the marine environment: a review.</a> Marine pollution bulletin. 62 (12), 2588-2597 (2011).</li><li>Zarfl, C., 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=Microplastics+in+oceans.">Microplastics in oceans.</a> Marine Pollution Bulletin. 62, 1589-1591 (2011).</li><li>Hanke, G., 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=.">.</a> MSFD GES technical subgroup on marine litter. Guidance on monitoring of marine litter in European Seas. , (2013).</li><li>L&#246;der, M. G. J., Gerdts, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methodology+used+for+the+detection+and+indentification+of+microplastics+-+A+critical+appraisal.">Methodology used for the detection and indentification of microplastics - A critical appraisal.</a> Marine anthropogenic litter. , 201-227 (2015).</li><li>Kang, J. H., Kwon, O. Y., Lee, K. W., Song, Y. K., Shim, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marine+neustonic+microplastics+around+the+southeastern+coast+of+Korea.">Marine neustonic microplastics around the southeastern coast of Korea.</a> Marine pollution bulletin. 96 (1), 304-312 (2015).</li><li>Lusher, A. L., Tirelli, V., O'Connor, I., Officer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microplastics+in+Arctic+polar+waters:+the+first+reported+values+of+particles+in+surface+and+sub-surface+samples.">Microplastics in Arctic polar waters: the first reported values of particles in surface and sub-surface samples.</a> Scientific reports. 5, (2015).</li><li>Shu, J. -. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+Marangoni+waves+due+to+impulsive+motion+of+a+submerged+body.">Transient Marangoni waves due to impulsive motion of a submerged body.</a> International Applied Mechanics. 40 (6), 709-714 (2004).</li><li>Rabaud, M., Moisy, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ship+wakes:+Kelvin+or+Mach+angle.">Ship wakes: Kelvin or Mach angle.</a> Physical Review Letters. 110 (21), 214503 (2013).</li><li>Hidalgo-Ruz, V., Gutow, L., Thompson, R. C., Thiel, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microplastics+in+the+marine+environment:+a+review+of+the+methods+used+for+identification+and+quantification.">Microplastics in the marine environment: a review of the methods used for identification and quantification.</a> Environmental science &amp; technology. 46 (6), 3060-3075 (2012).</li><li>L&#246;der, M. G. J., Kuczera, M., Mintenig, S., Lorenz, C., Gerdts, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+plane+array+detector-based+micro-Fourier-transform+infrared+imaging+for+the+analysis+of+microplastics+in+environmental+samples.">Focal plane array detector-based micro-Fourier-transform infrared imaging for the analysis of microplastics in environmental samples.</a> Environmental Chemistry. 12 (5), 563-581 (2009).</li><li>Ioakeimidis, C., 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+degradation+potential+of+PET+bottles+in+the+marine+environment:+An+ATR-FTIR+based+approach.">The degradation potential of PET bottles in the marine environment: An ATR-FTIR based approach.</a> Scientific reports. 6, 23501 (2016).</li><li>McDermid, K. J., McMullen, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+analysis+of+small-plastic+debris+on+beaches+in+the+Hawaiian+archipelago.">Quantitative analysis of small-plastic debris on beaches in the Hawaiian archipelago.</a> Marine pollution bulletin. 48 (7), 790-794 (2004).</li><li>Eriksen, M., 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=Microplastic+pollution+in+the+surface+waters+of+the+Laurentian+Great+Lakes.">Microplastic pollution in the surface waters of the Laurentian Great Lakes.</a> Marine pollution bulletin. 77 (1-2), 177-182 (2013).</li></ol>

The authors have nothing to disclose.

由蝠净海面上塑料微粒采样为海面上塑料微粒的采样一种广泛使用的方法，但迄今还没有统一的方法。大体积的水可通过蝠网进行过滤，从而捕集塑料微粒的相关数的可能性高，结果被认为是可靠的。不同的样品中结果的可比性是通过规范化放心。在我们的情况下，该浓度通过由净开口的水平宽度拖网距离乘以相关的取样区。另一种选择是使用一个流量计，固定在网的开口。使用流量计是可能的，因为净曼塔其侧翼是海面上非常稳定，因此在跳浪是最小的。流量计记录的过滤水的体积，从而使每水样16卷结果的正常化。 <p class="jove_content"&gt;最常用蝠网具有大约300微米的筛目大小和3 - 4.5米长。这些尺寸进行优化，以避免网的堵塞，并允许一定量的水的采样尽可能大。拖网速度推荐为介于2 - 结3，但它是依赖于波高度，风速和海流。这是很重要的，该蝠网是在监督下采样期间的全部时间，如果它开始跳频，在拖网速度必须减小。该拖网时间，建议大约30分钟，而是取决于悬浮物的浓度。它可能发生浮游物有时会堵塞蝠网。在这种情况下，拖网，必须立即停止，否则微型塑料颗粒可能丢失和净可能会损坏。蝠网是最经常从容器的侧面固定。这也是最合适的选择，而蝠鲼网肯定是出了醒区。在一些调查曼塔网从船的船尾固定17，18，但你必须要确保网是出来之后区域的情况下。的距离，在其上拖网设置为取样，应单独确定，因为所引起的容器湍流的区域从容器的尺寸和从船上19，20的速度而变化。 从海面样本的微型颗粒分离是最常见的视觉识别21刚刚完成。颗粒大于1毫米可以很容易地用肉眼识别，而颗粒小于1mm需要使用体视显微镜。以减少混淆非塑料颗粒与塑料的，使用上立体显微镜偏振的光的可能性，建议。塑料颗粒误认的可能性变得更小颗粒的高。由此颗粒&gt; 0.5，MM可以仅通过使用立体显微镜来识别视觉上21。为颗粒小于0.5mm额外的，更精确的方法是必需的如微ATR-FTIR光谱仪21。 在塑料微粒分离来自样品的处理样品污染与空气长丝的可能性非常高。出于这个原因，控制悬空在工作台上培养皿强烈建议潜在的污染物空气中颗粒物的鉴定。即，数据的质量强烈依赖于:1）用样品，2）的质量和立体显微镜的放大倍数，以及3）的有机物质的量的样品16中工作的人的精度。视觉识别后，强烈建议进行分析排序的颗粒为材料8的化学鉴定可用的技术之一。 几种方法聚合物鉴定存在，其中FTIR光谱和拉曼光谱是最frequenTLY使用22。红外和拉曼光谱是互补的技术以及其准确性是相似的。在我们的协议中，红外光谱和显微红外光谱与"衰减全反射"（ATR）介绍。它们是使用简单，它们能够快速和准确的结果。塑料聚合物具有与不同的带图案高度特异性的红外（IR）光谱，从而使红外光谱的最佳技术用于塑料微粒21的识别。与样品，使特征IR光谱22的测量交互时的IR辐射的能量激励一个特定的分子的振动。红外光谱还可提供关于颗粒，如氧化23和降解24的水平的强度的额外信息。而ATR-FTIR适合较大颗粒的化学识别（&gt; 0.5mm）的，微ATR-FTIR光谱可以提供关于颗粒＆＃的化学结构信息60 0.5毫米，因为它结合了显微镜和红外光谱仪的功能。 使用FTIR和微FTIR光谱之前，微型塑料颗粒必须预先干燥的，因为水强烈吸收IR辐射22，和纯化的，在情况下，它们都覆盖着生物膜和/或其它的有机和无机粘剂，其可以影响红外光谱。以纯化样品的大多数非侵入性的方式是通过搅拌，并用新鲜水25漂洗。如果这还不够，则建议使用30％的过氧化氢。所有其它的方法可对微型塑料颗粒的负面影响（如超声清洗可以进一步细分颗粒，强酸性或碱性溶液可能会损坏几个塑料聚合物等），因此不推荐使用。更有前途的是采用了连续酶促消化，为塑料友好纯化步骤。使用不同技术的酶纯化（例如脂肪酶，mylase，蛋白酶，几丁质酶，纤维素酶，蛋白酶K）已成功地应用于减少浮游生物的生物基质，从而被证明是一个有价值的技术，以尽量减少在红外光谱测量22矩阵文物。 通过视觉识别和选出的颗粒的化学鉴定塑料微粒的分离都非常耗时的过程。这项工作必须由一个准确和耐心的人谁具有立体显微镜，不仅在识别塑料粒子，而且在识别生物物质的经验来进行。即使是有经验的人无法从甲壳素或硅藻片段22歧视明确所有潜在的微型颗粒。因此，视觉分拣的误差率范围为20％26至70％21，并与减小粒径增大。

Microplastic pollution in the sea represents a growing concern to contemporary society, due to the constant increase in plastic production and its subsequent disposal and accumulation in the marine environment1. Even if plastic macro litter would no longer enter the seas, microplastic pollution would continue to grow due to fragmentation of already existing plastic litter in the sea2. The majority of microplastic pollution studies were carried out in marine and fresh water ecosystems and mainly addressed sea surface pollution3.
The term microplastic refers to plastic particles smaller than 5 mm in size4. This term describes a heterogeneous mixture of particles, which can differ in size (from a few microns to several millimeters), color and shape (from very different shapes of fragments to long fibers). Microplastic particles can be of a primary or secondary origin5. Microplastic of primary origin is manufactured as small particles used in the cosmetics industry (pilling cr&#232;me etc.) or chemical industry as precursor for other plastic products (e.g. plastic pellets used in plastic industry). Microplastic of secondary origin arise via the degradation of larger plastic pieces in the environment due to physical and chemical processes, induced by light, heat, oxygen, water and organisms6. In 2015, four types of microplastic sources were defined: larger plastic litter, cleaning products, medicines and textiles6. The main source (80 %) of larger plastic litter is assumed to be land based7. Microplastic from cosmetic products, medicines and textile enters water ecosystems through sewage and storm waters6. Microplastic particles most frequently found in water ecosystems are fragments from larger plastic litter and textile fibers8. 
Microplastics have several negative effects on the environment. Their small size allows them to enter the food web through ingestion by marine organisms9, 10. Ingested particles can cause physical damage or block the digestive system of animals11. Particles can also be carriers of persistent organic pollutants (POPs). Their hydrophobic surface and favorable ratio of large surface area to small volume, enables POPs to adsorb onto the microplastics12. In the environment or digestive systems of animals who ingest them, POPs and other plastic additives can be leached from microplastic particles13. 
Previous studies reported the ubiquitous presence of microplastics in the marine environment3, from the water column to the bottom sediments. The threat of microplastic pollution was already identified by the Marine Strategy Framework Directive in the EU and, consequently, mandatory monitoring of microplastics was advised14. Accordingly, the EU Technical Subgroup on Marine Litter (TSG-ML) prepared recommendations for monitoring of microplastics in the European seas15. Thus, the video guidelines for microplastics sampling are of high importance, as they support comparative monitoring and a coherent management process all over the world.
This protocol was developed within the DeFishGear project for the first monitoring of microplastic pollution in the Adriatic Sea. Recommendations from the document "Guidance on Monitoring of Marine Litter in European Seas" by TSG-ML15 were taken into account. This protocol describes the methodology for microplastics sampling on the sea surface, separation of microplastics from the samples, and chemical analysis of microplastic particles to confirm that particles are from plastic material and to identify the type of plastic. Sampling was done by the use of a manta net, which is the most suitable equipment for sampling in calm waters16. Separation of microplastics from the samples was carried out by visual identification using a stereomicroscope. Isolated particles were later chemically identified using Fourier transform infrared (FTIR) spectroscopy and micro FTIR spectroscopy.

<table><tbody><tr><td>In this protocol no specific equipment or reagents were used.</td><td></td><td></td><td></td></tr></tbody></table>

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.

所描述的协议的第一个结果是根据其视觉特征（见表1）分为六类微型塑料颗粒。第一类，且通常是最丰富的，是片段（图1）。他们是刚性的，厚厚的，用锋利的边缘弯曲和不规则的形状。它们可以在各种不同的颜色。第二类是膜（图2）。他们同时出现在不规则形状，但与片段相比，他们很瘦，灵活，通常是透明的。第三类是颗粒（图3），一般从塑料工业始发。它们是不规则的，圆的形状，并且通常在尺寸更大，约5毫米的直径。它们通常是在一侧平坦，并且可以是各种颜色。第四类是颗粒（图4）。在与粒料相比较，它们具有规则圆形形状，并且通常更小的尺寸，大约1毫米的直径。它们出现在自然的色彩（白色，米色，褐色）。第五类是丝（图5）。它们是，毗邻的片段，最丰富的类型的微型颗粒。它们可以是短或长，具有不同的厚度和颜色。最后一类是泡沫（图6）。他们最常来自发泡胶的大颗粒。它们是柔软的，形状不规则和白色至黄色。 塑料微粒取样和样品分析的主要结果是每个样品的微型颗粒的数目。这些数据可以是每平方公里进一步规范。用于标准化的计算公式为: 每个样品/采样区域的微型颗粒， （图;图7表2,3），其中取样区被由蝠净的开口的宽度采样距离乘以计算的。此外，颗粒可与即时进行分析年龄分析软件。结果包括每个粒子（表4）的最大长度和面积。图8a显示的图像分析之前颗粒和图8b是图像分析，其中每个粒子测量和编号之后。最后，建议每个样品颗粒的总或最高可能数量的化学分析。使用傅立叶变换红外光谱的选定颗粒的光谱获取，如图上图9这个频谱然后与从软件库（图10）的频谱进行比较。最后的结果将显示，如果一个给定的粒子是塑料或不与指示的塑料从化学结构的类型。 <table fo:keep-together.within-page="1"><tbody><tr><td> 1 </td><td>片段</td></tr><tr><td> 2 </td><td>电影</td></tr><tr><td> 3 </td><td>小球</td></tr><tr><td> 4 </td><td>颗粒</td></tr><tr><td>五</td><td>花丝</td></tr><tr><td> 6 </td><td>泡沫<html>小号</td></tr></tbody></table> 表1:微型塑料颗粒的类别。 <img alt="图1" src="/files/ftp_upload/55161/55161fig1.jpg" /> 图1:片段:从类别颗粒的例子。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/55161/55161fig2.jpg" /> 图2:电影:从类别颗粒的例子。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> /55161fig3.jpg"/&gt; 图3:颗粒:从类别颗粒的例子。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/55161/55161fig4.jpg" /> 图4:从类颗粒的实施例:粒剂。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/55161/55161fig5.jpg" /> 图5:从类颗粒的实施例:长丝。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图6:泡沫:从类别颗粒的例子。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <table fo:keep-together.within-page="1"><tbody><tr><td>采样距离[公里] </td><td> 2 </td></tr><tr><td>曼塔宽度[公里] </td><td> 0.0006 </td></tr><tr><td>取样面积[公里2] </td><td> 0.0012 </td></tr></tbody></table> 表2:从调查数据，用于每平方公里的微型颗粒的计算的实施例 。 <table fo:keep-together.within-page="1"><tbody><tr><td></td><td> 没有 </td><td> 无/ 平方公里 </td></tr><tr><td>片段 </td><td> 301 </td><td> 250833 </td></tr><tr><td> 影片 </td><td> 45 </td><td> 37500 </td></tr><tr><td> 小球 </td><td> 15 </td><td> 12500 </td></tr><tr><td> 颗粒 </td><td> 8 </td><td> 6667 </td></tr><tr><td> 泡沫 </td><td> 33 </td><td> 27500 </td></tr><tr><td> 花丝 </td><td> 223 </td><td> 185833 </td></tr></tbody></table> 表3:从调查，其中该分类的数据分为6组进行计数和每公里 2 标准化结果的实施例 （没有-粒子数）。 <img alt="图7" src="/files/ftp_upload/55161/55161fig7.jpg" /> 图7:的对视觉分类后，代表性的结果示例文章（无 - 粒子数）。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <table fo:keep-together.within-page="1"><tbody><tr><td> 区域指数 </td><td> 区域[平方毫米] </td><td> 最大长度（mm） </td></tr><tr><td> 1 </td><td> 8.010 </td><td> 5.506 </td></tr><tr><td> 2 </td><td> 10.517 </td><td> 5.628 </td></tr><tr><td> 3 </td><td> 12.185 </td><td> 5.429 </td></tr><tr><td> 4 </td><td> 3.367 </td><td> 3.367 </td></tr><tr><td>五</td><td> 2.475 </td><td> 2.155 </td></tr><tr><td> 6 </td><td> 1.809 </td><td> 2.943 </td></tr><tr><td> 7 </td><td> 6.604 </td><td> 5.238 </td></tr><tr><td> 8 </td><td> 5.779 </td><td> 4.037 </td></tr><tr><td> 9 </td><td> 4.472 </td><td> 3.791 </td></tr><html><tr><td> 10 </td><td> 16.907 </td><td> 5.355 </td></tr><tr><td> 11 </td><td> 7.246 </td><td> 3.733 </td></tr><tr><td> 12 </td><td> 7.867 </td><td> 4.622 </td></tr><tr><td> 13 </td><td> 64.11 </td><td> 5.056 </td></tr><tr><td> 14 </td><td> 3.281 </td><td> 3.070 </td></tr><tr><td> 15 </td><td> 12.937 </td><td> 5.554 </td></tr><tr><td> 16 </td><td> 6.709 </td><td> 3.716 </td></tr></tbody></table> 表4:其中区域[毫米 2]和各颗粒的最大长度（mm）进行测量 的图像分析结果的实施例 。 <img alt="图8" src="/files/ftp_upload/55161/55161fig8.jpg" /> 图8:图像的实施例之前获得的a）和b）用图像分析软件颗粒图像分析后。ecsource.jove.com/files/ftp_upload/55161/55161fig8large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <img alt="图9" src="/files/ftp_upload/55161/55161fig9.jpg" /> 图9:对选定的颗粒测得的标记峰和它们的波长[-1]一个光谱的实施例。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig9large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图10" src="/files/ftp_upload/55161/55161fig10.jpg" /> 图10:从选定的粒子从ATR-FTIR光谱库最佳匹配取得的频谱进行比较的例子。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55161/55161fig10large.jpg" target="_blank">请点击此处查看该F的放大版本</a>igure。

Watch this Scientific Journal Video about Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis at JoVE.com

Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

以下协议描述的方法:海面上的塑料微粒采样，颗粒的微型和化学鉴定分离。该协议是由MSFD技术小组对海洋垃圾公布了塑料微粒监测的建议。

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 ...

protocol-for-microplastics-sampling-on-sea-surface-sample

Nanyang Technological University

Research

JoVE Journal

Environment

49.3K 次观看.  Institute for water of the Republic of Slovenia. 以下协议描述的方法:海面上的塑料微粒采样，颗粒的微型和化学鉴定分离。该协议是由MSFD技术小组对海洋垃圾公布了塑料微粒监测的建议。 

视频: Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

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

Here we introduce a series of thoroughly tested and well standardized research protocols adapted for use in remote marine environments. The sampling protocols include the assessment of resources available to the microbial community (dissolved organic carbon, particulate organic matter, inorganic nutrients), and a comprehensive description of the viral and bacterial communities (via direct viral and microbial counts, enumeration of autofluorescent microbes, and construction of viral and microbial metagenomes). We use a combination of methods, which represent a dispersed field of scientific disciplines comprising already established protocols and some of the most recent techniques developed. Especially metagenomic sequencing techniques used for viral and bacterial community characterization, have been established only in recent years, and are thus still subjected to constant improvement. This has led to a variety of sampling and sample processing procedures currently in use. The set of methods presented here provides an up to date approach to collect and process environmental samples. Parameters addressed with these protocols yield the minimum on information essential to characterize and understand the underlying mechanisms of viral and microbial community dynamics. It gives easy to follow guidelines to conduct comprehensive surveys and discusses critical steps and potential caveats pertinent to each technique.

Here, we present a comprehensive protocol to assess the organic and inorganic nutrient availability and the abundance and structure of microbial and viral communities in remote marine environments.

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

Clean Sampling and Analysis of River and Estuarine Waters for Trace Metal Studies

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination control. Developments of some techniques aiming to reduce trace metal contamination in the last couple of decades have resulted in concentrations reported now being orders of magnitude lower than those in the past. These low concentrations often necessitate preconcentration of water samples prior to instrumental analysis of samples. Since contamination can appear in all phases of trace metal analyses, including sample collection (and during preparation of sampling containers), storage and handling, pretreatments, and instrumental analysis, specific care needs to be taken in order to reduce contamination levels at all steps. The effort to develop and utilize "clean techniques" in trace metal studies allows scientists to investigate trace metal distributions and chemical and biological behavior in greater details. This advancement also provides the required accuracy and precision of trace metal data allowing for environmental conditions to be related to trace metal concentrations in aquatic environments.
This protocol that is presented here details needed materials for sample preparation, sample collection, sample pretreatment including preconcentration, and instrumental analysis. By reducing contamination throughout all phases mentioned above for trace metal analysis, much lower detection limits and thus accuracy can be achieved. The effectiveness of "clean techniques" is further demonstrated using low field blanks and good recoveries for standard reference material. The data quality that can be obtained thus enables the assessment of trace metal distributions and their relationships to environmental parameters.

Special care using "clean techniques" is required to properly collect and process water samples for trace metal studies in aquatic environments. A protocol for sampling, processing, and analytical procedures with the aim of obtaining reliable environmental monitoring data and results with high sensitivity for detailed trace metal studies is presented.

清洁取样和痕量金属研究河和河口水域分析

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination ...

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 ...

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 ...

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 ...

Sampling, Sorting, and Characterizing Microplastics in Aquatic Environments with High Suspended Sediment Loads and Large Floating Debris

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only recently have microplastics in freshwater systems, such as rivers and lakes, been quantified. Microplastic sampling at the surface usually consists of deploying drift nets behind either a stationary or moving boat, which limits the sampling to environments with low levels of suspended sediments and floating or submerged debris. Previous studies that employed drift nets to collect microplastic debris typically used nets with &#8805;300 &#181;m mesh size, allowing plastic debris (particles and fibers) below this size to pass through the net and elude quantification. The protocol detailed here enables: 1) sample collection in environments with high suspended loads and floating or submerged debris and 2) the capture and quantification of microplastic particles and fibers &lt;300 &#181;m. Water samples were collected using a peristaltic pump in low-density polyethylene (PE) containers to be stored before filtering and analysis in the lab. Filtration was done with a custom-made microplastic filtration device containing detachable union joints that housed nylon mesh sieves and mixed cellulose ester membrane filters. Mesh sieves and membrane filters were examined with a stereomicroscope to quantify and separate microplastic particulates and fibers. These materials were then examined using a micro-attenuated total reflectance Fourier transform infrared spectrometer (micro ATR-FTIR) to determine microplastic polymer type. Recovery was measured by spiking samples using blue PE particulates and green nylon fibers; percent recovery was determined to be 100% for particulates and 92% for fibers. This protocol will guide similar studies on microplastics in high velocity rivers with high concentrations of sediment. With simple modifications to the peristaltic pump and filtration device, users can collect and analyze various sample volumes and particulate sizes.

Most microplastic research to date has occurred in marine systems where suspended solid levels are relatively low. Focus is now shifting to freshwater systems, which may feature high sediment loads and floating debris. This protocol addresses collecting and analyzing microplastic samples from aquatic environments that contain high suspended solid loads.

高悬浮泥沙荷载和大型漂浮碎屑水生环境中 Microplastics 的取样、分类和表征

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only ...

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 ...

Seawater Sampling and Collection

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass concentration, nucleic acid purification, cell abundance, nutrient and trace gas analyses. For today's demonstration samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. An A-frame derrick, with a multi-purpose winch and cable system, is used in combination with Niskin or Go-Flo water sampling bottles. Conductivity, Temperature, and Depth (CTD) sensors are also used to sample the underlying water mass. To minimize outgassing, trace gas samples are collected first.  Then, nutrients, water chemistry, and cell counts are determined. Finally, waters are collected for biomass filtration. The set-up and collection time for a single cast is ~1.5 hours at a maximum depth of 215 meters. Therefore, a total of 6 hours is generally needed to complete the collection series described here.

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass concentration, nucleic acid purification, cell abundance, nutrient and trace gas analyses.

海水采样和收集

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass ...

生物学

Small Volume (1-3L) Filtration of Coastal Seawater Samples

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For today s demonstration samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. This video documents small volume (~1 L) filtration of microbial biomass from the water column. The protocol is an extension of the large volume sampling protocol described earlier, with one major difference: here, there is no pre-filtration step, so all size classes of biomass are collected down to the 0.22 &mu;m filter cut-off. Samples collected this way are ideal for nucleic acid analysis. The set-up, filtration, and clean-up steps each take about 20-30 minutes. If using two peristaltic pumps simultaneously, up to 8 samples may be filtered at the same time. To prevent biofilm formation between sampling trips, all filtration equipment must be rinsed with dilute HCl and deionized water and autoclaved immediately after use.

Small Volume 1-3L Filtration of Coastal Seawater Samples

This video documents small volume (~1 L) filtration of microbial biomass from the water column.

沿海海水样品的过滤体积小（1 - 3L）

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For today s ...

Sampling, Identification and Characterization of Microplastics Release from Polypropylene Baby Feeding Bottle during Daily Use

Microplastics (MPs) are becoming a global concern due to the potential risk to human health. Case studies of plastic products (i.e., plastic single-use cups and kettles) indicate that MP release during daily use can be extremely high. Precisely determining the MP release level is a crucial step to identify and quantify the exposure source and assess/control the corresponding risks stemming from this exposure. Though protocols for measuring MP levels in marine or freshwater has been well developed, the conditions experienced by household plastic products can vary widely. Many plastic products are exposed to frequent high temperatures (up to 100 &#176;C) and are cooled back to room temperature during daily use. It is therefore crucial to develop a sampling protocol that mimics the actual daily-use scenario for each particular product. This study focused on widely used polypropylene-based baby feeding bottles to develop a cost-effective protocol for MP release studies of many plastic products. The protocol developed here enables: 1) prevention of the potential contamination during sampling and detection; 2) realistic implementation of daily-use scenarios and accurate collection of the MPs released from baby feeding bottles based on WHO guidelines; and 3) cost-effective chemical determination and physical topography mapping of MPs released from baby feeding bottles. Based on this protocol, the recovery percentage using standard polystyrene MP (diameter of 2 &#181;m) was 92.4-101.2% while the detected size was around 102.2% of the designed size. The protocol detailed here provides a reliable and cost-effective method for MP sample preparation and detection, which can substantially benefit future studies of MP release from plastic products.

This study detailed a reliable and cost-effective protocol for microplastics collection and detection from the daily use of plastic products.

每日使用期间从聚丙烯婴儿喂养瓶中释放的微塑料的采样、鉴定和特征

Microplastics (MPs) are becoming a global concern due to the potential risk to human health. Case studies of plastic products (i.e., plastic single-use ...