We would like to thank Dr. Nicholas J. Ashbolt and Dr. G. Shay Fout for their review of the manuscript.

1.准备玻璃器皿和过滤器外壳<ol style="margin-left: 40px;"><li>除非另有说明，所有消毒玻璃器皿，过滤器外壳和溶液在121℃下进行15分钟。为了保证无菌，覆盖所有开口或暴露的表面或者用铝箔或胶带固定前消毒纸。 </li><li>通过将过滤器壳（47毫米直径）以1 L侧臂Erlenmeyer烧瓶装配过滤装置。收集所需的过滤器：直径47 mm正电/负电盘式过滤器。 </li><li>制备1L的1.5％牛肉提取物（凝聚;产生的絮凝物颗粒时溶液pH值降低至3.5，或不絮凝;其中不聚集在pH 3.5），用0.05M甘氨酸，溶解15克牛肉提取物中的1升的去离子水并加入3.75克甘氨酸。 注：非絮凝牛肉提取物将需要叔胺浓度，然后加入硅藻土。 </li><li>加入洗脱添加剂钠polyphosphate牛肉提取物在0.1％的浓度。 </li><li>添加1M盐酸溶液，逐滴，以混合牛肉提取物，pH值溶液至所需的pH值。蒸压牛肉提取物在121℃15-30分钟。 </li><li>衡量0.1克精细，中型或大型粒子硅藻土（与唯一的非絮凝牛肉膏使用）的。 </li><li>通过附加兼容油管锥形侧臂烧瓶中，并于真空出口准备真空/抽吸。 </li></ol> 2.准备解决方案和病毒股票<ol style="margin-left: 40px;"><li>制备1升的1M HCl中。 </li><li>制备1升的1M NaOH处理。 </li><li>制备的1×PBS溶液（137 mM氯化钠，2.7 mM的氯化钾，1.47毫K 2 PO 4，和4.3毫摩尔的NaH 2 PO 4）。 <ol><li>调节pH的1×PBS的9.0使用1M的NaOH。 </li></ol></li><li>准备和稀释股票的病毒，以10 4 -10 5最可能数（MPN）毫升-1混合1毫升股票病毒的9毫升的PBS（pH值7），如先前所描述的8,13。储存在-80℃下在1ml等分试样。 <ol><li>消毒病毒的股票通过注射器过滤（0.22微米孔径），以消除潜在的污染物。 </li></ol></li><li>脱氯自来水<ol><li>措施1升用无菌无菌自来水量筒并倒入含有磁性搅拌棒的2升烧杯中。 </li><li>加入将0.7克的硫代硫酸钠并混合直到颗粒溶解。如果使用1M HCl或1M的NaOH溶液根据需要调整pH值的自来水至7-7.5。 </li><li>解冻的病毒刺突（1毫升）中，并添加穗总体积为1升的无菌脱氯自来水。混合10分钟。 </li><li>制备方法空白（无腺病毒脱氯自来水）与每个实验和过程中相同的方式加标样品。 </li></ol></li></ol> 3.制备微孔过滤装置<ol style="margin-left: 40px;"><li>删除无菌占地过滤器外壳。保证过滤器外壳上碗，下滤网之间适当的配合在过滤过程中，避免样品泄漏。 </li><li>从1升锥形瓶中取出无菌覆盖。确保过滤器单元的软木紧密地配合在锥形瓶中的顶部开口，创造了良好的密封。 </li><li>从较低的过滤网拆下顶部碗过滤单元，并将47毫米阳电滤盘正视屏幕上用无菌镊子。安装过滤器外壳碗底部的屏幕并锁定到位逆时针扭动。 </li></ol> 4.自来水过滤（主浓度） <ol style="margin-left: 40px;"><li>取100mL病毒的尖刺用无菌量筒自来水。 </li><li>倒入百毫升病毒飙升自来水到含有无论是47毫米折叠玻璃/纤维素过滤器或纳米氧化铝/玻璃纤维盘滤清器滤芯外壳。 </li><li>让水慢慢穿过过滤器。适用于温和真空去除剩余的残留物Ø从过滤面F的自来水。 </li><li>如果使用胰蛋白酶处理，加入5 ml 0.25％胰蛋白酶的过滤表面并孵育5分钟。 </li><li>从用50毫升无菌去离子水过滤表面清洗胰蛋白酶。 </li></ol>从过滤器5.病毒洗脱<ol style="margin-left: 40px;"><li>从2升Erlenmeyer烧瓶中除去过滤器壳体，并放置到250毫升侧臂烧瓶中。 </li><li>取100mL牛肉膏的使用量筒。慢慢倒入100毫升牛肉膏进入过滤器外壳。 </li><li>允许牛肉提取倒通过过滤器，捕获牛肉提取在一个小的（250毫升）的Erlenmeyer烧瓶中。运用真空过滤从通过残余的牛肉膏拉。 </li></ol> 6.硅藻土二级浓度<ol style="margin-left: 40px;"><li>传输瓶中含有洗脱病毒的轰动板。加灭菌磁力搅拌棒并将其放置在搅拌盘，搅拌创造轻微的漩涡。 </li><li>添加0.1克的硅藻​​土粉末至100毫升牛肉提取物，并允许硅藻土分散。将无菌的pH探头放入牛肉膏。 </li><li>添加逐滴1 M盐酸至牛肉膏实现pH值4.0。允许缓慢混合10分钟。 </li><li>放置使用的2L Erlenmeyer烧瓶47毫米预过滤到过滤器外壳（如前所述）。 </li><li>在预过滤倒入百毫升牛肉膏/硅藻土混合。让牛肉膏倒过。适用于轻微的真空过滤器，从通过剩余牛肉膏拉。 </li><li>将金属管夹上的过滤器外壳喷结束。附加的无菌15mL的聚丙烯管中收集到金属管夹。放置过滤器壳体回Erlenmeyer烧瓶。 </li><li>加入5毫升的1×PBS缓冲液，pH 9.0经硅藻土上收集的预过滤器。 </li><li>让PBS滴通过预过滤器。适用于轻微的真空通过残余的PBS拉成15个ml收集管。 </li></ol> 7.有机絮凝二级浓度<ol style="margin-left: 40px;"><li>传输含病毒洗脱的烧杯，以小的轰动板。加灭菌磁力搅拌棒至100毫升牛肉提取物和混合创造轻微的漩涡。 </li><li>将无菌的pH探头放入牛肉膏。 </li><li>添加1M的HCl滴至牛肉提取物和调节pH值至3.5。拌30分钟。 </li><li>倾洗脱液于250毫升的离心锥形管和离心机在2500×g离心15分钟，在4ºC。 </li><li>倒掉上清，注意不要不要破坏沉淀。重悬在5ml的1×PBS，pH值9的颗粒。 </li><li>离心该悬浮液在4000×g离心10分钟，在4ºC。倒掉上清弃沉淀。 </li><li>将pH调节至7-7.5，过滤消毒通过0.22微米注射器式滤器并冷冻在-80℃。 </li></ol> 8.病毒核酸提取<ol style="margin-left: 40px;"><li>提取根据制造商的说明，用适当的市售试剂盒的病毒核酸。 </li></ol> &lt;P类=“j​​ove_title”&gt; 9。腺病毒4​​0/41定量PCR <ol style="margin-left: 40px;"><li>使用引物和探针，以及反应混合物和热循环曲线所公布8,13。 注：检测化验的估计上限是每反应约5 qPCR的单位。 </li><li>添加的PCR扩增混合物组分的浓度如下：10毫摩尔Tris（pH值8.3），50mM的氯化钾，4.5毫的MgCl 2，10毫的dNTPs，10μM引物和1μM的探针和0.5微升聚合酶以便在45最终卷微升可以加入到孔中的适当数量的96孔PCR反应板内。 </li><li>负载45微升的PCR扩增混合物中，96孔，快速反应的PCR板的每个相应井。 </li><li>加入5微升DNA提取样本的PCR板的合适的孔中。 </li><li>捂盘与耐热盖密封，旋转盘移动任何液滴PCR板孔底部。 </li><li>设置PCR扩增循环的FOLL流入的：95℃10分钟，随后为15秒的40个循环的95℃和60℃进行1分钟。 </li></ol> 10.数据分析<ol style="margin-left: 40px;"><li>为了确定病毒颗粒的最大可能数（MPN）625稀释范围，如前所述8,13：分析跨越1的样本的序列5倍稀释液的一式三份测量：5至1。 </li><li>为了确定病毒颗粒的最大可能数（MPN）15625稀释范围：腺病毒实验尖峰跨越1的串行5倍稀释液分析：5到1。 </li><li>使用的EPA MPN计算器（http://www.epa.gov/nerlcwww/online.html），调整计算器占稀释的类型（1：5或1:10），进行稀释的数量，以及数复制样品连续稀释。 </li><li>执行日志中的数据，随后归一体积的单位（ 例如 1毫升，100毫升， 等 10变换</EM&gt;）。除以存在于样品的病毒颗粒的数目计算回收率为给定的实验集中与存在于实验尖峰病毒颗粒的数目。 </li><li>评估不同实验变量的影响（ 例如，不同的硅藻土的类型，pH值范围，不同的滤波器类型和不同的次级浓缩技术）上的AdV回收率通过执行参数或非参数统计测试（ 例如 ，配对t检验，一个或两个往来港澳ANOVA），根据数据的分布和实验设计。 </li></ol>

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No competing financial interests. 
The views expressed in this article are those of the authors and do not necessarily reflect the views or policies of the U.S. Environmental Protection Agency. Mention of trade names or commercial products does not constitute endorsement or recommendation for use.

阳电过滤器是集中在水的病毒有效;然而，这些过滤器可以在它们的结构和组合物，其可以反过来改变其效力不同。私了这个问题，衣壳的结构和收费病毒株需要浓缩技术进行定制，以确保最佳恢复15之间变化。通过对现有的浓度的技术（ 例如，正电性的过滤器，牛肉提取物洗脱），靶病毒更有效浓度可以达到16,17简单的修改。 研究迄今为止通常集中于?...

人类肠道病毒是水源性疾病1-3中重要病原体，但一般存在于被污染的环境水低的数字，使得它们的检测困难而不浓度。用于浓缩病毒程序通常包括一个过滤步骤中的过滤器的洗脱液中，接着过滤洗脱，和仲浓度。一个常见的过滤方法依赖于使用荷电膜，如正电的过滤器（最近在4,5审查）的。这些过滤器依赖于捕捉的病毒悬浮在水中使用的过滤表面（正电荷）和靶向的病毒​​颗粒之间的静电相互作用（带负电）。两个正电的过滤器是商业上可用的依赖于该技术中，玻璃/纤维素和纳米氧化铝/玻璃纤维过滤器。该玻璃/​​纤维素滤器成本高达10倍，该纳米氧化铝/玻璃纤维的，其限制使用的玻璃/ CE的llulose过滤器进行例行病毒监控。最近的研究已经得出结论差异是名义在从室温水中6,7肠道病毒的恢复这两个滤波器之间，证明使用一种更便宜的过滤器的替代。其他过滤选项，例如带负电和玻璃毛过滤器进行了研究，然而，他们要么需要源水（电负性过滤器）的预处理或没有市售的（玻璃毛过滤器）。的病毒浓度程序的发展主要集中在以提高病毒回收率从水优化初级浓缩技术（过滤器）。然而，次级浓度程序，其中降低洗脱液的体积通常为1升至毫升卷，也可对病毒回收率8显著影响。 的肠道病毒次要浓度通常依赖于絮凝剂如一些类型的牛肉提取物（有机floccuLATION）或脱脂奶絮凝9-12从过滤器表面除去病毒颗粒。最近，利用再加上另外硅藻土（细颗粒）的牛肉膏另一二次浓度过程已表明承诺回收腺病毒，肠道病毒，诺如病毒和8,13,14。根据相似的原理，以在该病毒颗粒中的有机絮凝方法的硅藻土的浓度的作品附加到并通过改变悬浮液的pH释放从粒子（絮凝物或硅藻土）。这两个次级浓缩技术之间的比较中尖刺腺病毒（ADV）类型40和41 8的恢复已被评估。该研究的结论是，两个次级浓缩技术人腺病毒的回收统计学相似。然而，有机絮凝方法需要30分钟。孵育在pH 3.5，而硅藻土技术需要一个较短的温育（10分钟），在pH 4.0。有机flocculat离子也需要使用昂贵的实验室设备（离心机）叔浓缩过程中收集的絮凝物颗粒，与此相反的硅藻土技术仅使用基本的实验室设备（真空过滤），以硅藻土颗粒从悬浮液中分离出来。 过滤器和二次洗脱技术的某些组合也可影响病毒的回收率。一项研究的结论是，初级（正电的过滤器）的某些组合和次级浓缩技术（硅藻土或有机絮凝）的 ​​腺病毒13的回收的显著影响。这些结果表明，优化使用这些技术时，需要以最佳地从给定的水基质回收靶病毒。优化是一个耗时的，艰苦的过程许多研究者积极避免，因为无数的变量进行评估（过滤器类型/品牌，pH值洗脱液，硅藻土/有机絮凝）。 对于这种Study，一个步骤，开发了使用加标人腺病毒株40和41可以推测，以确定病毒浓度的最佳条件，从水，由于每个病毒类型显示了独特的衣壳形态和特定的衣壳电荷浓度的协议可能需要对每个病毒进行优化目标，以达到最佳的病毒回收。本研究为腺病毒40和41浓度的方法：1）评估病毒回收率自来水使用阳电过滤盘之后2）建立有机絮凝法对硅藻土技术作为辅助的浓度评价，以及3）评估洗脱缓冲液为大专浓度。

<table border="0" cellpadding="0" cellspacing="0" width="786">
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			<td height="21" style="height:21px;width:231px;">Hydrochloric Acid</td>
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</table>

a small volume procedure for viral concentration from water

Small-scale concentration of viruses (sample volumes 1-10 L, here simulated with spiked 100 ml water samples) is an efficient, cost-effective way to identify optimal parameters for virus concentration. Viruses can be concentrated from water using filtration (electropositive, electronegative, glass wool or size exclusion), followed by secondary concentration with beef extract to release viruses from filter surfaces, and finally tertiary concentration resulting in a 5-30 ml volume virus concentrate. In order to identify optimal concentration procedures, two different electropositive filters were evaluated (a glass/cellulose filter [1MDS] and a nano-alumina/glass filter [NanoCeram]), as well as different secondary concentration techniques; the celite technique where three different celite particle sizes were evaluated (fine, medium and large) followed by comparing this technique with that of the established organic flocculation method. Various elution additives were also evaluated for their ability to enhance the release of adenovirus (AdV) particles from filter surfaces. Fine particle celite recovered similar levels of AdV40 and 41 to that of the established organic flocculation method when viral spikes were added during secondary concentration. The glass/cellulose filter recovered higher levels of both, AdV40 and 41, compared to that of a nano-alumina/glass fiber filter. Although not statistically significant, the addition of 0.1% sodium polyphosphate amended beef extract eluant recovered 10% more AdV particles compared to unamended beef extract.

<html>硅藻土选择三种不同类型的硅藻土进行之前表现最好的变体的选择进行测试。 Celites细到中等大小的粒子产生的最高腺病毒回收率。使用较大celites导致较低的回收率为AdV40和41（范围为32％-100％）（ 图1）。腺病毒4​​0的平均回收率为144％±52％（精细），115％±28％（中）和82％±53％（大）颗粒celites和AdV41，132％±39％（精细），83％±的25％（培养基）和50％±1...

Watch this Scientific Journal Video about A Small Volume Procedure for Viral Concentration from Water at JoVE.com

A Small Volume Procedure for Viral Concentration from Water

An approach was developed for identifying optimal viral concentration conditions for small volume water samples using spikes of human adenovirus. The techniques described here are used to identify concentration parameters for other viral targets, and applied to large-scale viral concentration experimentation.

小体积的方法从水中病毒浓度

Small-scale concentration of viruses (sample volumes 1-10 L, here simulated with spiked 100 ml water samples) is an efficient, cost-effective way to ...

a-small-volume-procedure-for-viral-concentration-from-water

Research

JoVE Journal

Environment

10.0K 次观看.  U.S. EPA. An approach was developed for identifying optimal viral concentration conditions for small volume water samples using spikes of human adenovirus. The techniques described here are used to identify concentration parameters for other viral targets, and applied to large-scale viral concentration experimentation. 

视频: A Small Volume Procedure for Viral Concentration from Water

Laboratory-determined Phosphorus Flux from Lake Sediments as a Measure of Internal Phosphorus Loading

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) loading from lake sediments can account for a substantial portion of the total P load in eutrophic, and some mesotrophic, lakes. Laboratory determination of P release rates from sediment cores is one approach for determining the role of internal P loading and guiding management decisions. Two principal alternatives to experimental determination of sediment P release exist for estimating internal load: in situ measurements of changes in hypolimnetic P over time and P mass balance. The experimental approach using laboratory-based sediment incubations to quantify internal P load is a direct method, making it a valuable tool for lake management and restoration.
Laboratory incubations of sediment cores can help determine the relative importance of internal vs. external P loads, as well as be used to answer a variety of lake management and research questions. We illustrate the use of sediment core incubations to assess the effectiveness of an aluminum sulfate (alum) treatment for reducing sediment P release. Other research questions that can be investigated using this approach include the effects of sediment resuspension and bioturbation on P release.
The approach also has limitations. Assumptions must be made with respect to: extrapolating results from sediment cores to the entire lake; deciding over what time periods to measure nutrient release; and addressing possible core tube artifacts. A comprehensive dissolved oxygen monitoring strategy to assess temporal and spatial redox status in the lake provides greater confidence in annual P loads estimated from sediment core incubations.

Lake eutrophication is a water quality issue worldwide, making the need to identify and control nutrient sources critical. Laboratory determination of phosphorus release rates from sediment cores is a valuable approach for determining the role of internal phosphorus loading and guiding management decisions.

实验室确定的磷通量湖泊沉积物作为内部磷载入一个测量

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) ...

科研

环境科学

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated (WATER-PAM) Fluorometry

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be subsampled throughout the experiment1&#8211;7. Subsampling of large volumes can be problematic for several reasons: 1) it causes variation in the total volume and the surface area:volume ratio of the culture during the experiment; 2) pseudo-replication (i.e., replicate samples from the same treatment flask8) is often employed rather than true replicates (i.e., sampling from replicate treatments); 3) the duration of the experiment is limited by the total volume; and 4) axenic cultures or the usual bacterial microbiota are difficult to maintain during long-term experiments as contamination commonly occurs during subsampling.
The use of microtiter plates enables 1 ml culture volumes to be used for each replicate, with up to 48 separate treatments within a 12.65 x 8.5 x 2.2 cm plate, thereby decreasing the experimental volume and allowing for extensive replication without subsampling any treatment. Additionally, this technique can be modified to fit a variety of experimental formats including: bacterial-algal co-cultures, algal physiology tests, and toxin screening9&#8211;11. Individual wells with an alga, bacterium and/or co-cultures can be sampled for numerous laboratory procedures including, but not limited to: WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry, microscopy, bacterial colony forming unit (cfu) counts and flow cytometry. The combination of the microtiter plate format and WATER-PAM fluorometry allows for multiple rapid measurements of photochemical yield and other photochemical parameters with low variability between samples, high reproducibility and avoids the many pitfalls of subsampling a carboy or conical flask over the course of an experiment.

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated WATER-PAM Fluorometry

The goal of this procedure is to demonstrate the reproducibility and adaptability of using a microtiter plate format for microalgal screening. This rapid screen combines WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry to measure photosynthetic yield as an indicator of Photosystem II (PSII) health with small volume bacterial-algal co-cultures.

小体积的生物测定评估细菌/浮游植物共培养使用水脉冲幅度调制（WATER-PAM）荧光

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be ...

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most experiments examine only two or a few levels of CA concentration and a single soil type, but if CA can be varied as a gradient from subambient to superambient concentrations on multiple soils, we can discern whether past ecosystem responses may continue linearly in the future and whether responses may vary across the landscape. The Lysimeter Carbon Dioxide Gradient Facility applies a 250 to 500 &#181;l L-1 CA gradient to Blackland prairie plant communities established on lysimeters containing clay, silty clay, and sandy soils. The gradient is created as photosynthesis by vegetation enclosed in in temperature-controlled chambers progressively depletes carbon dioxide from air flowing directionally through the chambers. Maintaining proper air flow rate, adequate photosynthetic capacity, and temperature control are critical to overcome the main limitations of the system, which are declining photosynthetic rates and increased water stress during summer. The facility is an economical alternative to other techniques of CA enrichment, successfully discerns the shape of ecosystem responses to subambient to superambient CA enrichment, and can be adapted to test for interactions of carbon dioxide with other greenhouse gases such as methane or ozone.

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

The Lysimeter Carbon Dioxide Gradient Facility creates a 250 to 500 &#181;l L-1 linear carbon dioxide gradient in temperature-controlled chambers housing grassland plant communities on clay, silty clay, and sandy soil monoliths. The facility is used to determine how past and future carbon dioxide levels affect grassland carbon cycling.

一个外线<sub&gt; 2</sub&gt;浓度梯度用于测试CO<sub&gt; 2</sub&gt;浓度与土壤对草地生态系统功能的影响

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most ...

A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and enables to perform rapid, automated and simple SPE. The pre-concentrated solution is compatible with analysis by immunoassay, with a low organic solvent content. A method is described for the extraction and pre-concentration of natural hormone 17&#946;-estradiol in 100 ml water samples. Reverse phase SPE is performed with octadecyl-silica sorbent and elution is done with 200 &#181;l of methanol 50% v/v. Eluent is diluted by adding di-water to lower the amount of methanol. After preparing manually the SPE column, the overall procedure is performed automatically within 1 hr. At the end of the process, estradiol concentration is measured by using a commercial enzyme-linked immune-sorbent assay (ELISA). 100-fold pre-concentration is achieved and the methanol content in only 10% v/v. Full recoveries of the molecule are achieved with 1 ng/L spiked de-ionized and synthetic sea water samples.

A protocol for the extraction and pre-concentration of estradiol from water samples by using an automated and miniaturized system is presented.

对水样的自动固相萃取小污染物的免疫学分析的简单方法

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and ...

Extracting Metrics for Three-dimensional Root Systems: Volume and Surface Analysis from In-soil X-ray Computed Tomography Data

Plant roots play a critical role in plant-soil-microbe interactions that occur in the rhizosphere, as well as processes with important implications to climate change and crop management. Quantitative size information on roots in their native environment is invaluable for studying root growth and environmental processes involving plants. X-ray computed tomography (XCT) has been demonstrated to be an effective tool for in situ root scanning and analysis. We aimed to develop a costless and efficient tool that approximates the surface and volume of the root regardless of its shape from three-dimensional (3D) tomography data. The root structure of a Prairie dropseed (Sporobolus heterolepis) specimen was imaged using XCT. The root was reconstructed, and the primary root structure was extracted from the data using a combination of licensed and open-source software. An isosurface polygonal mesh was then created for ease of analysis. We have developed the standalone application imeshJ, generated in MATLAB1, to calculate root volume and surface area from the mesh. The outputs of imeshJ are surface area (in mm2) and the volume (in mm3). The process, utilizing a unique combination of tools from imaging to quantitative root analysis, is described. A combination of XCT and open-source software proved to be a powerful combination to noninvasively image plant root samples, segment root data, and extract quantitative information from the 3D data. This methodology of processing 3D data should be applicable to other material/sample systems where there is connectivity between components of similar X-ray attenuation and difficulties arise with segmentation.

A methodology for obtaining visual and quantitative root structure information from X-ray computed tomography data acquired in-soil is presented.

从在土X射线计算机断层扫描数据量和表面分析：提取指标三维根系

Plant roots play a critical role in plant-soil-microbe interactions that occur in the rhizosphere, as well as processes with important implications to ...

Soil Lysimeter Excavation for Coupled Hydrological, Geochemical, and Microbiological Investigations

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled Earth-system processes. Such knowledge is imperative in improving predictions of hydro-biogeochemical cycles, especially under climate change scenarios. We present an experimental method, designed to capture sub-surface heterogeneity of an initially homogeneous soil system. This method is based on destructive sampling of a soil lysimeter designed to simulate a small-scale hillslope. A weighing lysimeter of one cubic meter capacity was divided into sections (voxels) and was excavated layer-by-layer, with sub samples being collected from each voxel. The excavation procedure was aimed at detecting the incipient heterogeneity of the system by focusing on the spatial assessment of hydrological, geochemical, and microbiological properties of the soil. Representative results of a few physicochemical variables tested show the development of heterogeneity. Additional work to test interactions between hydrological, geochemical, and microbiological signatures is planned to interpret the observed patterns. Our study also demonstrates the possibility of carrying out similar excavations in order to observe and quantify different aspects of soil-development under varying environmental conditions and scale.

This study presents an excavation method for investigating subsurface hydrological, geochemical, and microbiological heterogeneity of a soil lysimeter. The lysimeter simulates an artificial hillslope which was initially under homogeneous condition and had been subjected to approximately 5,000 mm of water over eight cycles of irrigation in an 18-month period.

土壤蒸渗仪开挖加之水文，地球化学和微生物调查

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled ...

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 ...

Capturing Flow-weighted Water and Suspended Particulates from Agricultural Canals During Drainage Events

The purpose of this study is to describe the methods used to capture flow-weighted water and suspended particulates from farm canals during drainage discharge events. Farm canals can be enriched by nutrients such as phosphorus (P) that are susceptible to transport. Phosphorus in the form of suspended particulates can significantly contribute to the overall P loads in drainage water. A settling tank experiment was conducted to capture suspended particulates during discrete drainage events. Farm canal discharge water was collected in a series of two 200 L settling tanks over the entire duration of the drainage event, so as to represent a composite subsample of the water being discharged. Imhoff settling cones are ultimately used to settle out the suspended particulates. This is achieved by siphoning water from the settling tanks via the cones. The particulates are then collected for physico-chemical analyses.

Nutrients present in particulate form can contribute significantly to the overall loads in agricultural drainage waters. This study describes a novel method to capture flow-weighted water and suspended particulates from farm canal drainage over the entire duration of the drainage event.

在排水过程中捕获农用运河中的流动加权水和悬浮颗粒

The purpose of this study is to describe the methods used to capture flow-weighted water and suspended particulates from farm canals during drainage ...

A Simple Planting Technique for Re-establishing Trees Where Frequent Inundation Occurs

Many of the Everglades tree islands have lost elevation over the past century and most of their trees have died such that they are now covered with herbaceous plants. This protocol describes a simple, cost-effective tree planting technique needed for restoring degraded Everglade tree islands. The design is patterned after a natural Everglades process that creates floating peat islands, which allows tree survival and growth in flooded conditions and often leads to the development of tree islands. Commercially available peat bags were used as the medium for the growth and establishment of potted native tree saplings. The pop-up configuration floated initially and provided additional elevation to minimize inundation, with a single native tree species sapling and a single tree fertilizer spike. During a 3 year study involving 105 pop-ups, most plants survived (80%) and many thrived. Determining whether this technique can establish trees on a degraded tree island will require longer studies and extensive field tests.

This protocol describes a simple, cost-effective tree planting technique for restoring degraded Everglades tree islands that experience inundation. The design creates an island (pop-up) which floats initially and adds elevation to promote tree survival and growth under flooded conditions.

常淹发生的重构树简易造林技术

Many of the Everglades tree islands have lost elevation over the past century and most of their trees have died such that they are now covered with ...

Optimized Procedure for Determining the Adsorption of Phosphonates onto Granular Ferric Hydroxide using a Miniaturized Phosphorus Determination Method

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide (GFH), with little effort and high reliability. The phosphonate, e.g., nitrilotrimethylphosphonic acid (NTMP), is brought into contact with the GFH in a rotator in a solution buffered by an organic acid (e.g., acetic acid) or Good buffer (e.g., 2-(N-morpholino)ethanesulfonic acid) [MES] and N-cyclohexyl-2-hydroxyl-3-aminopropanesulfonic acid [CAPSO]) in a concentration of 10 mM for a specific time in 50 mL centrifuge tubes. Subsequently, after membrane filtration (0.45 &#181;m pore size), the total phosphorus (total P) concentration is measured using a specifically developed determination method (ISOmini). This method is a modification and simplification of the ISO 6878 method: a 4 mL sample is mixed with H2SO4 and K2S2O8 in a screw cap vial, heated to 148-150 &#176;C for 1 h and then mixed with NaOH, ascorbic acid and acidified molybdate with antimony(III) (final volume of 10 mL) to produce a blue complex. The color intensity, which is linearly proportional to the phosphorus concentration, is measured spectrophotometrically (880 nm). It is demonstrated that the buffer concentration used has no significant effect on the adsorption of phosphonate between pH 4 and 12. The buffers, therefore, do not compete with the phosphonate for adsorption sites. Furthermore, the relatively high concentration of the buffer requires a higher dosage concentration of oxidizing agent (K2S2O8) for digestion than that specified in ISO 6878, which, together with the NaOH dosage, is matched to each buffer. Despite the simplification, the ISOmini method does not lose any of its accuracy compared to the standardized method.

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide, with little effort and high reliability. In a buffered solution, the phosphonate is brought into contact with the adsorbent using a rotator and then analyzed via a miniaturized phosphorus determination method.

用小型化磷测定法测定膦酸在粒状氢氧化铁上吸附的优化方法

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric ...