这项研究部分由美国国家科学基金会的快速程序（NSF-1048925，1048919，和1048914）艾莉森Keimowitz，明郭李笃Okeke，和詹姆斯·桑德斯的支持。

1.设备的研制<ol><li>核心内衬的制备<ol><li>计算将使用卷来获得核心片=πR2×厚度厚度;最终的体积必须&lt;50 立方厘米 。用10cm的纤芯直径，可以得到2厘米厚的片。 注意:这是没有必要具有体积是一个完整的50毫升，但得到的孔隙水体积将成比例越小。 </li><li>使用线锯（或类似的）切片一个芯衬垫，或相同直径的塑料管，成2厘米的环（或其它的厚度，如果使用不同的直径芯）。获得3-5环。 </li><li>清洁所有的塑料材料会在接触沉积物，包括核心套，瓶盖核心，戒指，核心切片机，离心管，注射器，和一次性勺子。 （浸泡的塑料材料，在10％HCl中24小时后，冲洗塑料材料nanopure（22兆欧）水3倍，并且允许材料在空气中干燥，优选在层流罩，包装前）。 </li></ol></li><li>杰克实验室制备<ol><li>切下一块胶合板的，采用线锯，以覆盖实验室千斤顶的顶板的6"×6"的大小，并在使用空穴这个夹板的中心钻一个孔锯安装到钻。 注意:这个洞应该只是比PVC扩片直径略大;对于这里使用的尺寸，孔应该是2.75"。 </li><li>钻四个在此夹板的边缘小孔上使用一个手电钻或钻床常规钻头以匹配实验室千斤顶的顶板上的孔。拧紧夹板用扎带实验室插孔。 注:应该有这个夹板没有"回旋"。 </li></ol></li><li>钻在约2&#39;×1.5&#39;胶合板比使用孔锯芯衬垫的外径稍大的中心的孔。对于直径10厘米的核心班轮，一个10.5-11.5厘米的洞会appropria德。这种木材是核心指导板。 </li><li>芯挤压机的制备（活塞） <ol><li>获得完全吻合核心衬里内的橡胶塞的实验室。如果没有可用的，用剃刀刮胡子之一大小。不要使用过小的瓶塞。 </li><li>螺杆实验室塞子约1&#39;-1.5"长，1"直径的木钉或用扫帚柄，与防水绝缘胶带塞子的面盖螺丝头。 注意:动器的较大的面应在销了。 </li></ol></li><li>切PVC管的约1.5&#39;长成〜6"长部分。 注:内部直径应比在步骤1.4.2销钉较大，但比在步骤1.4.1止动小。这个协议假定使用标准2"的PVC的2.375真正的外径。"这些PVC片是核心扩展。 </li><li>装配在材料清单中列出的所有其他必要的设备。 注:此SH乌尔德在家庭实验室中收集并带​​到野外实验室。 </li></ol> 2.设置场站实验室<ol><li>建立手套袋<ol><li>夹紧芯引导板的工作表面（台面，实验室表面等 ）。确保为核心班轮孔是在工作面上，而且是开放的。 </li><li>将在核心导向板一次性手套包，并运行从手套包管道进入到N 2坦克的调节器上的管道。确保燃油箱是用气缸板凳钳牢牢固定。 </li><li>大盘油管手套袋由环绕周围袋用电工胶带管道的外部封住这个入口点。确保约8"管延伸入手套袋的内部的管夹滑到手套袋内的管道;离开该夹紧开放。 </li><li>在手套袋在底部切一个"X"孔用一个盒子刀或直剃刀的核心指导板。这个x应大于芯衬垫直径小。 </li><li>加载与表1中找到的项的手套袋中。 </li></ol></li><li>建立核心<ol><li>将实验室插孔，其中手套包贴在工作区下方的地板上。放置芯通过在手套袋的底部，在步骤2.1.4的"X"切，保持在直立位置。 注:约4-6"的核心应该扩展核心稳定板之上。 </li><li>保持核心稳定，让研究人员2至2.2.6执行步骤2.2.3。 </li><li>磁带核心的塑料手套袋围绕"X"，用大量的密封性好电工胶带。 </li><li>插入，芯挤压机的手柄成"直径，〜6"2 PVC长垫片，随后〜3"长耦合，接着另一个垫片。继续这种模式，直到PVC完全盖手柄;它可以延伸短的距离近的手柄的端部。放置芯挤压机（与PVC间隔件和联接器）的核心的底部的下方。 </li><li>支持核心挤出机实验室千斤顶使挤出机可以支持的核心。此时，保持插座尽可能低，并使用核心隔板尽可能。 </li><li>使用一箱刀围绕底部核心帽精心切割。该切口应当导致芯盖的环留围绕芯衬套的外部和芯盖的代替对芯材的平坦圆形部分。 </li><li>将手放入手套袋（研究员1）。持芯稳定，使得它不会在芯稳定板之上更进一步移动，而研究员2开始转动实验室千斤顶慢慢提高的核心。核心挤出机应该进入核心班轮，并开始推泥沙像向上推弹出。 </li><li>扭动芯挤压机侧小幅如果东北侧EDED将其插入到所述芯衬垫（研究员2）。对于轻微的流行，当它进入做好准备。 </li><li>继续提高芯直到芯材料的顶部处或附近的芯衬垫的顶部（研究者1和2）。请注意，核心上限应在此过程中保持在芯的顶上。 </li></ol></li><li>密封袋手套<ol><li>仔细检查所有必需的生活用品在手套袋;打开便携式氧计。 </li><li>开所有离心管，水瓶，以及包含截留的空气的其他项目。如果核心有积水上面顶空，打开顶盖以清除这个顶部空间。 </li><li>打开调节器，使氮的通过管的流动到手套袋以中等速度。一般一个在所有阀门打开为宜最后调节器级〜15磅的压力。 注:这应该是速度不够快，感觉就像皮肤上强风，但不是那么强大，它驱散手套袋SUP层数。 </li><li>点入氮气手套袋的各个领域，并推动了氮的主要开放的最好的一个人的能力。关掉氮气。 </li><li>通过重复步骤2.3.6至2.3.9三次吹扫手套袋中三次。 注意:一个人可以做到这一点，但它可能对两个人更容易，使人们可以保持他或她的手在属于手套袋的一部分的手套。 </li><li>通过围绕其1-2蹦极绳索几次封袋手套的主要开幕。打开氮气的类似流如在2.3.3。地方研究员1的手在手套袋内置手套。 </li><li>从袋的一个区域移动至氮气管到另一个填充氮气的手套袋中。点管子插入任何缝隙，如打开离心管中，水在上面喷瓶， 等等。 </li><li>通过关闭管夹关掉氮气。注意，这可以在不除去水库来完成earcher 1的手从手套包。 </li><li>打开手套袋的前面，并使用研究者1的身体，推尽可能多的气体从袋子尽可能的。 注:包应该在这一点上是它里面的周边耗材持平。 </li><li>填充手套袋到舒适的压力和使用的管夹关掉氮气。一些试验和错误可能需要找到舒适的压力，因为太满，这是难以移动一个人的手臂，而太空，这是很难看到并操纵对象。打开短期内重新填充袋，如果它似乎有一个缓慢的泄漏或越来越以任何理由空虚氮管夹。打开包的前面，让出少量的氮气，如果它变得不舒服满。 </li><li>检查溶解氧在便携式氧计的水平。它应该是在1％以下。 </li><li>将手套袋手套到一次性手套。这将提高灵活性。改变他们时，他们GEŧ污渍或损坏;丢弃在袋中的废弃物容器中。 </li></ol></li></ol> 3.切片核心注:该过程的这个部分更容易与两位研究者来完成。 <ol><li>除去使用注射器积水。注射器过滤这种水并放入50ml离心管中。 </li><li>删除一个核心部分， <ol><li>上方放置芯衬垫的顶部的芯切片环。提高核心起来（第3.3节），直到沉淀物的顶部是在环的顶部。 </li><li>插入芯衬垫和环的顶部之间的芯切片机。沉积物部分现在坐的核心切​​片机顶上。 </li><li>移动使用一次性勺子沉积物至50ml管中。帽管紧，一旦它充满。 </li><li>冲洗核心切片机和核心切片环与超纯水;喷脏废为手套袋内废物容器。一次性勺子也可以冲洗，或丢弃，这取决于所需要的数量。擦干核心切片机切片核心环纸巾。 </li><li>重复步骤3.2直到没有进一步的芯材依然存在。 </li></ol></li><li>提高核心<ol><li>贯穿3.2有必要提高的核心。在使用实验室千斤顶小的增量执行此操作。执行以下三个步骤一次实验室千斤顶完全展开。 </li><li>握住手套袋（研究员1）代替的核心。 </li><li>放下实验室插孔最低设置，当它被完全压缩（研究员2）。确保核心是与手套包和研究员2的双手从下方支持在研究1的手的地方举行。 </li><li>填充芯挤压机的底端和与PVC间隔件和管件的插孔之间的空间。确保核心是由核心扩展让核心的顶部走之前牢固地支撑。 </li></ol></li></ol> 4.惠tracting孔隙水<ol><li>打开手套袋并取出离心管的机架。 </li><li>根据需要通过消灭纸巾，水等空废物容器内取出任何土壤，液体或冷凝清理杂物袋。 </li><li>用蹦极绳索松散重新密封手套袋中。 </li><li>权衡其现在包含剖开的沉积物（如需要）离心管中。 </li><li>包住离心管的顶部用绝缘带密封的盖/管交界处。 </li><li>用胶带后称重来平衡离心管。添加或删除电工胶带至0.5g中得到的权重。 注意:通常情况下，从芯部的顶部管会比从底部打火机。 </li><li>在离心机和离心以最大加速度地方管（推荐1100 XG）20分钟。 注意:较高的利率离心将允许孔隙水的分离较大。 </li><li>从centrifu删除电工胶带戈管或者通过unpeeling或通过与之前的管返回到手套袋中的剃刀切割。返回离心管离心后机架，并返回到手套袋中。 </li><li>添加另一组离心管，注射器和注射器过滤器到手套袋中。这些新的管将持有孔隙水和可预先标记。 </li><li>再次清除手套塑料袋，在步骤2.3.5。如果手套袋只简单地打开，取出管，未清洁的，这将是可接受的清除它只有两次。如果正在使用氧气监视器，保证在手套袋中的气氛&lt;1％O 2。不要忘了打开新添加的离心管和清除它们。 </li><li>双手插在手套袋手套;和以前一样，请用一次性手套。 </li><li>打开一管。从使用注射器沉积物上方取出孔隙水，然后用注射器过滤器附加到注射器的尖端。如果孔隙水和沉积物很好的分离，porewat器可以直接倒入取下筒与上尖端的注射器过滤器的注射器。 </li><li>通过注射过滤器推水到适当的离心管中。 注:将需要一些力量;可能需要每个样品多于一个针筒过滤器。 </li><li>替换在含有沉积物和含有孔隙水管内的离心管的盖子。 </li><li>对每个样品重复步骤4.13 4.15通过。 </li><li>打开手套袋中，取出样品。既沉积物和孔隙水管可以重新称重。 </li><li>如果需要立即分析孔隙水。分析可包括（但不限于）为主要离子11离子色谱，菲洛嗪铁形态12中 ，砷的形态由伏安13和硫化物形态14。 </li><li>在冷冻干燥托运人孔隙水和沉积物（约-80℃）保存形态为在适当分析后为计划的分析。据米也AY适当样本保持在密闭容器中或氮气吹扫镀铝袋冷却。 </li><li>再利用的手套袋中的第二芯如果需要的话（在2.2.2开始）。两个芯后，将手套袋中通常需要改变。 </li></ol>

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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectrophotometric+Determination+of+Hydrogen+Sulfide+in+Natural+Waters.">Spectrophotometric Determination of Hydrogen Sulfide in Natural Waters.</a> L&amp;O. 14, 454-458 (1969).</li></ol>

本文所描述的技术是一种灵活的，能够为各种各样的位置，芯尺寸，芯部厚度等 ，有此系统的三个主要组分进行调整。 首先，准备正确尺寸的芯挤压系统，用于将待分析的核心。这里给出的说明假设一个大约30"芯;长得多芯可能需要更多的PVC增量剂件和PVC管件完全挤出如本领域的更正是困难得多管理计划的挤压系统和仔细的包装，。 其次，确保手?...

研究人员常常希望来研究沉积物 - 水系统的氧化还原状态和地质微生物学。这理想地利用来自两个沉积物和孔隙水的数据，孔隙水通常是系统的敏感显示器和是一个公共源，虽然不是唯一的源，生态暴露于氧化还原敏感性重金属1如砷和铀。孔隙水数据可以原位使用扩散平衡滤波器，也称为"皮珀斯，"安装到沉积物2来获得。偷窥者都在外地网站之前开始现场工作，并在那里过的很长一段时间多次访问可以到野外现场进行， 例如 Shotyk 3闻名设置最常用的。因此，许多情况下不允许使用皮珀斯，如站点只有很短的时间内可以访问或者获得多个探样品，以确定进一步的调查应该发生4。此外皮珀斯不同时采样沉积物水样采集。 当希望采样沉淀物和水一起，或在该领域网站，偷窥安装是不可行的，最常用的方法，以获得沉淀物和水是沉积物核化。获得未混合芯是一个关键的前体在此工作5描述的过程。一旦获得芯孔隙水可以通过挤压6或离心来获得;都具有优点和缺点。离心通常被认为是提取沉积物岩芯，7孔隙水虽然必须小心，以防止或沉积物孔隙水的氧化最可靠的方法。 在这种方法中，我们描述的核心挤出和离心提取孔隙水以最小的氧化。作者使用在各种上下文包括海洋8的本文中所描述的方法中，被污染的湖泊<sup&gt; 9，和湿地10。示出的具有代表性的数据表明，在还原条件得以保存。用离心机之外，使用的材料是廉价的，而且这种方法可以适用于各种各样的地球化学和地球微生物研究问题。

<table border="0" cellpadding="0" cellspacing="0" width="811">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;Disposable glove bag(s).&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>Z106089-1EA</td>
			<td>&#160;One per two cores to be processed is usually sufficient.</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">&#160;N2 tank</td>
			<td>Praxair</td>
			<td></td>
			<td>Often gas supply companies can deliver these directly to the field laboratory.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nitrogen gas regulator</td>
			<td>VWR</td>
			<td>55850-478</td>
			<td>Or similar</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Several feet of tubing that fits the regulator</td>
			<td>VWR</td>
			<td>89403-862</td>
			<td>Or similar</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Safety equipment to secure the tank</td>
			<td>VWR</td>
			<td>60142-006</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Adjustable tubing clamp</td>
			<td>VWR</td>
			<td>62849-112</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Waterproof, good sealing electrical tape</td>
			<td>Scotch</td>
			<td>Super 33+</td>
			<td>Widely available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;2-4 short bungee cords</td>
			<td></td>
			<td></td>
			<td>Widely available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Squirt bottles of nanopure water</td>
			<td>VWR</td>
			<td>16650-082</td>
			<td>Any similar bottle is fine; pack an additional supply of nanopure water to refill these.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Large supply of paper towels and kimwipes.</td>
			<td></td>
			<td></td>
			<td>Widely available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">50 mL centrifuge tubes</td>
			<td>VWR</td>
			<td>21008-951</td>
			<td>Acid cleaned as described in protocol.&#160; At least 2/core section needed.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Several permanent in markers.</td>
			<td></td>
			<td></td>
			<td>Widely available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;Several straight razor blades and box cutters.</td>
			<td></td>
			<td></td>
			<td>Widely available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Centrifuge</td>
			<td>Beckman-Coulter</td>
			<td>Allegra X-22</td>
			<td>Faster rotor allows greater separation.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rotor to accommodate 50 mL tubes</td>
			<td>Beckman-Coulter</td>
			<td>SX-4250</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">]50 mL plastic syringes without black rubber tip on the barrel</td>
			<td>VWR</td>
			<td>66064-764&#160;</td>
			<td>Acid cleaned as described in protocol.&#160; At least 1/core section needed, plus 1 for overlying water.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringe filters compatible with aqueous solutions.</td>
			<td>VWR</td>
			<td>28143-310&#160;</td>
			<td>Either 0.45 &#956;m or 0.20 &#956;m poresizes may be used.&#160; Plan on five filters per core section processed.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Plastic (disposable) spoons.</td>
			<td></td>
			<td></td>
			<td>Widely available; Acid cleaned as described in protocol.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Several boxes of disposable gloves.</td>
			<td></td>
			<td></td>
			<td>Widely available</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">Large plastic beakers or other waste containers to place in the glove bag.</td>
			<td>VWR</td>
			<td>13890-148</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Laboratory balance</td>
			<td>VWR</td>
			<td>10205-008</td>
			<td>An available balance will be fine; high precision not required</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dry shipper, pre-charged with liquid nitrogen</td>
			<td>VWR</td>
			<td>82005-416</td>
			<td>Needed only if samples are being returned to the home laboratory for sensitive analyses.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Laboratory notebooks</td>
			<td></td>
			<td></td>
			<td>Water repellent can be useful</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Core liners</td>
			<td>Watermark</td>
			<td>77280</td>
			<td>Available from Forrestry Suppliers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Core caps</td>
			<td>Ben Meadows</td>
			<td>218105</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Core slicers</td>
			<td>McMaster Carr</td>
			<td>8707K111</td>
			<td>Cut this into 9 3x3 squares</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PVC spacers</td>
			<td>McMaster Carr</td>
			<td>48925K96</td>
			<td>Cut this into short lengths</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PVC couplings</td>
			<td>McMaster Carr</td>
			<td>4880K76</td>
			<td>Approximately 12 needed</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dowel</td>
			<td></td>
			<td></td>
			<td>Widely available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lab stopper</td>
			<td>VWR</td>
			<td>59580-400</td>
			<td>Check to ensure the correct size to fit snugly within the core liners</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Plywood for core guidance plate and top of lab jack</td>
			<td></td>
			<td></td>
			<td>Widely available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lab jack</td>
			<td>VWR</td>
			<td>89260-826</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Clamps</td>
			<td></td>
			<td></td>
			<td>Widely available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Portable oxygen monitor</td>
			<td>RKI instruments</td>
			<td>OX-07</td>
			<td></td>
		</tr>
	</tbody>
</table>

sediment core sectioning and extraction of pore waters under anoxic conditions

我们演示了切片沉积岩芯和提取孔隙水，同时保持无氧状态的方法。一个简单，廉价的系统构建，可以运到一个临时的工作空间接近现场采样网站（），以方便快速分析。芯被挤压成一个便携式手套袋中，在那里它们被切片并每1-3厘米厚的部分（取决于芯直径）被密封到50ml离心管中。孔隙水用离心分离的手套袋中的外面，然后返回到手套袋用于从沉淀分离。这些提取出来的孔隙水样品可立即进行分析。氧化还原敏感的物种，如硫化物，铁形态，和砷的形态的直接分析表明孔隙水的氧化最小;一些样品显示没有可检测的Fe（III）还原物种的约100％， 例如 100％的Fe（Ⅱ）。这两种沉积物和孔隙水样可以保存到主一旦返回到实验室覃用于进一步分析的化学物质。

<html>类型获得的结果取决于所执行的分析和对从其获得芯地球化学设置。溶解氧可以在所提取的孔隙水进行测量，但在很多情况下，这将是下面的芯的前几个厘米为零。通常提供更多有意义的信息的分析包括铁形态（铁II / III铁）12，砷的形态（如III /作为V）13，和硫化物14。还原物质如硫化物的存在指示既还原环境和足够缺氧芯切片期间维持和孔?...

Watch this Scientific Journal Video about Sediment Core Sectioning and Extraction of Pore Waters under Anoxic Conditions at JoVE.com

Sediment Core Sectioning and Extraction of Pore Waters under Anoxic Conditions

A protocol for sectioning sediment cores and extracting pore waters under anoxic conditions in order to permit analysis of redox sensitive species in both solids and fluids is presented.

沉积物核心切片和孔隙水萃取缺氧条件下

We demonstrate a method for sectioning sediment cores and extracting pore waters while maintaining oxygen-free conditions. A simple, inexpensive system is ...

sediment-core-sectioning-extraction-pore-waters-under-anoxic

Research

JoVE Journal

Environment

8.9K 次观看.  Vassar College. A protocol for sectioning sediment cores and extracting pore waters under anoxic conditions in order to permit analysis of redox sensitive species in both solids and fluids is presented.

视频: Sediment Core Sectioning and Extraction of Pore Waters under Anoxic Conditions

Determination of Microbial Extracellular Enzyme Activity in Waters, Soils, and Sediments using High Throughput Microplate Assays

Much of the nutrient cycling and carbon processing in natural environments occurs through the activity of extracellular enzymes released by microorganisms. Thus, measurement of the activity of these extracellular enzymes can give insights into the rates of ecosystem level processes, such as organic matter decomposition or nitrogen and phosphorus mineralization. Assays of extracellular enzyme activity in environmental samples typically involve exposing the samples to artificial colorimetric or fluorometric substrates and tracking the rate of substrate hydrolysis. Here we describe microplate based methods for these procedures that allow the analysis of large numbers of samples within a short time frame. Samples are allowed to react with artificial substrates within 96-well microplates or deep well microplate blocks, and enzyme activity is subsequently determined by absorption or fluorescence of the resulting end product using a typical microplate reader or fluorometer. Such high throughput procedures not only facilitate comparisons between spatially separate sites or ecosystems, but also substantially reduce the cost of such assays by reducing overall reagent volumes needed per sample.

Microplate based procedures are described for the colorimetric or fluorometric analysis of extracellular enzyme activity. These procedures allow for the rapid assay of such activity in large numbers of environmental samples within a manageable time frame.

在使用高通量微孔板测定水，土壤，沉积物和微生物测定胞外酶活性的

Much of the nutrient cycling and carbon  processing in natural environments occurs through the activity of extracellular  enzymes released by ...

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 ...

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 ...

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in nutrient and gas cycles. After cores with intact sediment-water interfaces are collected, they are submerged in incubation tanks and kept under aerobic conditions at in situ temperatures. To initiate a time course of overlying water chemistry, cores are sealed without bubbles using a top cap with a suspended stirrer. Time courses of 4-7 sample points are used to determine the rate of sediment water exchange. Artificial illumination simulates day-time conditions for shallow photosynthetic sediments, and in conjunction with dark incubations can provide net exchanges on a daily basis. The net measurement of N2 is made possible by sampling a time course of dissolved gas concentrations, with high precision mass spectrometric analysis of N2:Ar ratios providing a means to measure N2 concentrations. We have successfully applied this approach to lakes, reservoirs, estuaries, wetlands and storm water ponds, and with care, this approach provides valuable information on biogeochemical balances in aquatic ecosystems.

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

Here, we present small core incubations for the measurement of sediment-water gas and solute exchange. These will provide reliable measurements of sediment-water exchange that assess the role of sediment in influencing biological and biogeochemical processes in aquatic ecosystems.

Ø的底栖交易所<sub&gt; 2</sub&gt;，N<sub&gt; 2</sub&gt;和营养盐使用小的核心孵化

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in ...

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester fabric and glue can be folded and glued to construct a strip of compartments that house seeds and identifying information, while allowing contact with soil leachate, water, microorganisms, and ambient temperature. Strips may be constructed with a wide range of compartment numbers and sizes and allow the researcher to house a variety of genotypes within a single species, different species, or seeds that have experienced different treatments. As opposed to individual seed packets, strips are more easily retrieved as a unit. While replicate packets can be included within a strip, different strips can act as blocks or can be retrieved at different times for observation of seed behavior over time. We used a high temperature glue gun to delineate compartments and sealed the strips once the seed and tags identifying block and removal times were inserted. The seed strips were then buried in the field at the desired depth, with the location marked for later removal. Burrowing animal predators were effectively excluded by use of a covering of metal mesh hardware cloth on the soil surface. After the selected time interval for burial, strips were dug up and seeds were assessed for germination, dormancy and mortality. While clearly dead seeds can often be distinguished from ungerminated living ones by eye, dormant seeds were conclusively identified using a standard Tetrazolium chloride colorimetric test for seed viability.

Here we present a protocol for assessing seed survivorship, germination and dormancy under field conditions using buried, labeled seed strips and tetrazolium chloride (TZ) viability testing.

对于在野外条件下评估种子萌发，休眠和死亡率可靠的方法

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

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 ...

Extraction and Characterization of Surfactants from Atmospheric Aerosols

Surface-active compounds, or surfactants, present in atmospheric aerosols are expected to play important roles in the formation of liquid water clouds in the Earth&#39;s atmosphere, a central process in meteorology, hydrology, and for the climate system. But because specific extraction and characterization of these compounds have been lacking for decades, very little is known on their identity, properties, mode of action and origins, thus preventing the full understanding of cloud formation and its potential links with the Earth&#39;s ecosystems.
In this paper we present recently developed methods for 1) the targeted extraction of all the surfactants from atmospheric aerosol samples and for the determination of 2) their absolute concentrations in the aerosol phase and 3) their static surface tension curves in water, including their Critical Micelle Concentration (CMC). These methods have been validated with 9 references surfactants, including anionic, cationic and non-ionic ones. Examples of results are presented for surfactants found in fine aerosol particles (diameter &#60;1 &#956;m) collected at a coastal site in Croatia and suggestions for future improvements and other characterizations than those presented are discussed.

Methods are presented for the targeted extraction of surfactants present in atmospheric aerosols and the determination of their absolute concentrations and surface tension curves in water, including their Critical Micelle Concentration (CMC).

提取和表面活性剂的特性，从大气气溶胶

Surface-active compounds, or surfactants, present in atmospheric aerosols are expected to play important roles in the formation of liquid water clouds in ...

Continuous Instream Monitoring of Nutrients and Sediment in Agricultural Watersheds

Pollutant concentrations and loads in watersheds vary considerably with time and space. Accurate and timely information on the magnitude of pollutants in water resources is a prerequisite for understanding the drivers of the pollutant loads and for making informed water resource management decisions. The commonly used &#34;grab sampling&#34; method provides the concentrations of pollutants at the time of sampling (i.e., a snapshot concentration) and may under- or overpredict the pollutant concentrations and loads. Continuous monitoring of nutrients and sediment has recently received more attention due to advances in computing, sensing technology, and storage devices. This protocol demonstrates the use of sensors, sondes, and instrumentation to continuously monitor in situ nitrate, ammonium, turbidity, pH, conductivity, temperature, and dissolved oxygen (DO) and to calculate the loads from two streams (ditches) in two agricultural watersheds. With the proper calibration, maintenance, and operation of sensors and sondes, good water quality data can be obtained by overcoming challenging conditions such as fouling and debris buildup. The method can also be used in watersheds of various sizes and characterized by agricultural, forested, and/or urban land.

With the advancement of technology and the rise in end-user expectations, the need and use of higher temporal resolution data for pollutant load estimation has increased. This protocol describes a method for continuous in situ water quality monitoring to obtain higher temporal resolution data for informed water resource management decisions.

农业流域营养盐和泥沙的连续河道监测

Pollutant concentrations and loads in watersheds vary considerably with time and space. Accurate and timely information on the magnitude of pollutants in ...

Microfluidic Devices for Characterizing Pore-scale Event Processes in Porous Media for Oil Recovery Applications

Microfluidic devices are versatile tools for studying transport processes at a microscopic scale. A demand exists for microfluidic devices that are resistant to low molecular-weight oil components, unlike traditional polydimethylsiloxane (PDMS) devices. Here, we demonstrate a facile method for making a device with this property, and we use the product of this protocol for examining the pore-scale mechanisms by which foam recovers crude oil. A pattern is first designed using computer-aided design (CAD) software and printed on a transparency with a high-resolution printer. This pattern is then transferred to a photoresist via a lithography procedure. PDMS is cast on the pattern, cured in an oven, and removed to obtain a mold. A thiol-ene crosslinking polymer, commonly used as an optical adhesive (OA), is then poured onto the mold and cured under UV light. The PDMS mold is peeled away from the optical adhesive cast. A glass substrate is then prepared, and the two halves of the device are bonded together. Optical adhesive-based devices are more robust than traditional PDMS microfluidic devices. The epoxy structure is resistant to swelling by many organic solvents, which opens new possibilities for experiments involving light organic liquids. Additionally, the surface wettability behavior of these devices is more stable than that of PDMS. The construction of optical adhesive microfluidic devices is simple, yet requires incrementally more effort than the making of PDMS-based devices. Also, though optical adhesive devices are stable in organic liquids, they may exhibit reduced bond-strength after a long time. Optical adhesive microfluidic devices can be made in geometries that act as 2-D micromodels for porous media. These devices are applied in the study of oil displacement to improve our understanding of the pore-scale mechanisms involved in enhanced oil recovery and aquifer remediation.

The goal of this procedure is to easily and rapidly produce a microfluidic device with customizable geometry and resistance to swelling by organic fluids for oil recovery studies. A polydimethylsiloxane mold is first generated, and then used to cast the epoxy-based device. A representative displacement study is reported.

微流体器件用于表征多孔介质中的孔隙尺度事件过程, 用于采油应用

Microfluidic devices are versatile tools for studying transport processes at a microscopic scale. A demand exists for microfluidic devices that are ...

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 ...