我们感谢Khalied艾哈迈德，莱尼麦克奈特和德鲁Pinnix现场和实验室的援助，以及湖惠勒草坪现场实验室的支持人员，包括达斯汀·科贝特和马蒂教区，保持研究领域。这项工作是部分由北卡罗莱纳州立大学的中心草坪环境研究和教育的支持。

1.田间鉴定和确定<ol><li>确定研究领域代表选择的草坪制度。特别是考虑适当草种和土壤类型，以及包括生育，灌溉和修剪高度管理实践。 </li><li>进行叶面和土壤基质的初步残渣分析，以确保所关注农药（多个）是不可检测的。 </li><li>铺陈情节适当尺寸网站和样品采集方法，并纳入0.5-1.0米的小巷上积边界，允许人员客流量。 注:简介尺寸可以根据特定于站点的条件而变化，但至少应为3.3 平方米 。这将允许对一个3.7米×0.5米球行进通路，以及为脚的流量和植被取样等效相邻区域。 </li></ol> 2.启动试验<ol><li>二十四小时，以萌生刈草坪草的品种之前-system特定的高度和收集剪报。使用吹气球清除所有剩余的碎片和灌溉对土壤饱和。 注意:此时应灌溉允许在施药最佳湿度（ 即田间持水量）。 </li><li>在感兴趣的位点现在所登记的最大标记津贴选择农药施用量。施用率为特定产品和由美国环保署决定，并受法律上的所有农药的容器标签说明。 <ol><li>选择当前最小标记喷涂载体容量最大化叶面喷洒滞留。 </li></ol></li><li>在审判开始时，申请农药喷雾治疗，以图在气候条件有利于处理上保留植被。 注:适当的个人防护装备必须由喷头和nonapplicators试验区附近在治疗期间佩戴。始终保持nonapplicator人员上风喷涂应用。气候条件S于治疗植被有利于保持树冠包括水分（在应用干），风速（&lt;11公里/小时）和预测沉淀<ol><li>通过至少四个采样事件涵盖灌溉/降水期间地块。 注:地块被覆盖，以防止从处理过的植物农药洗成草坪茅草和/或土壤表面，使之少去除的和低估暴露风险。盖可通过材料和设计而变化，但应构造的无孔材料制成，以保持小区干燥并设计为使得处理过的植物不接触（例如，不直接铺设防水布上重复），这可能会危及随后农药转印采样。 </li></ol></li><li>每小时收集气象数据，包括空气温度，露点，相对湿度，日出时间，风向和风速在整个试验期间用数据解释提供帮助。 </li></ol> 3.质量控制样品<ol><li>科尔CT一个样本（50至100毫升）从研究中使用，确保农药产品的负荷率是正确的每种农药的容器。记录产品名称和每个容器的批号。 </li><li>收集通过从每个喷嘴收集输出施加各农药喷雾罐混合物的样品，使每罐混合样品（50至100毫升），以确保正确的喷雾溶液混合的过程。 </li><li>产生喷涂应用控件威尔士等人所述。 12，以确保农药的适当量被施加到图。 </li><li>作为威尔士等人生产领域强化和空白样品。 12在每个采样事件期间样本收集和储存过程量化耗散，并确定潜在的污染源。 注:设防样品浓度和溶剂可以基于化合物 - 和具体的分析考量。 </li></ol><ol><li>收集样品应用后立即，和再一次农药已在叶表面上干燥。取决于环境条件，这是应用后通常为1至3小时。 </li><li>继申请之日起，在采集样本，或者只是在日出后，当最大草坪草篷水分通常存在14。 <ol><li>确定样品的采集时序基于特定农药理化性质的应用程序之日起，与采样间隔减少农药持久性下降。时间表样品收集定时，使得在至少两个连续的nondetections通过实验完成发生。 注意:从先前的研究定量转移2,4-二甲基胺盐实施例（2,4-D;领域半衰期= 6.2天）:1）0天治疗后（DAT） - 喷雾后0小时2）0 DAT - 喷雾后1小时3）1 DAT; 4）2的DAT; 5）3 DAT; 6）6的DAT; 7）12 DAT; 8）24天14。 </li></ol></li></ol> 5.转换残留样品采集注:腈手套戴在所有样品收集程序，并且应尽可能经常需要防止污染被替换。 <img alt="图1" src="/files/ftp_upload/55182/55182fig1.jpg" /> 图1:手抹压量化。方法用来量化从fronthand（A）和反手（B）的压力擦拭动作。这样做，喷绿色食用色素溶液（水+ 10％体积/体积食用色素+ 1％体积/体积的非离子表面活性剂）上的无孔表面（例如，玻璃或金属盘），为用于农药打跑取样和按手。如由康皮奥等人所述通过数字图像分析量化的接触表面面积。 17 determi每个已知面积的图像绿色像素的NE的比例。收集来自预期的研究领域一个核心的草坪草在土壤在田间持水量，并将其固定数字体重秤。量化的向下的力时，手抹的质量。手抹压不应该超过2千帕。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55182/55182fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol><li> 手擦 注:需要一个人。 <ol><li>校准手擦拭压力为2帕。 <ol><li>通过计算在安全领域的土壤收集能力草坪一个核心的数字规模和紧迫的（按前去皮）150提供至200克擦力。 </li><li>量化用于草坪采样前台和反手的区域。主要使用手指而从此奠定了限制他们联系独特的草坪草的叶片。 <ol><li>制备10％（体积）的水基溶液/ v的绿色食用色素+ 1％体积/体积的非离子表面活性剂，并喷上的无孔表面以产生一个薄膜。 注:非离子表面活性是通过降低表面张力，增加解决方案覆盖了无孔的表面。 </li><li>与上一棉手套，按手无孔表面上（ 图1）。 </li><li>取出手套和通过接管每个手套的已知区域中的数字相片量化与无孔表面接触手的面积，并使用ImageJ的16选择用于绿色像素康皮奥等类似。 17。这将提供在绿色像素的总图象，它是用来计算面积的百分比。 </li></ol></li><li>结合擦力质量与擦拭面积，计算手的压力。 </li></ol></li><li>通过标记在地面上0.3米的距离校准手刮水速度1.2公里/小时调整手的速度，这样擦这个距离需要1秒。 </li><li>马克·山姆其中开口端朝下耦区域（420 cm 2）的通过将一个圆形桶（23厘米直径）倒置。施加轻微的压力，以桶的中心，并与周围的桶的外边缘相对手画（使用草皮划线漆）。 </li><li>用棉手套过上占主导地位的手丁腈手套，擦拭过30秒的采样区域。备用前端和反手表面，而垂直地调整方向（ 图2）。 <ol><li>重复步骤5.1.4用新棉手套。 注:在每个样品需手套变异可以基于感兴趣，草坪草系统和采样区域中的农药。这应该被事先确定为试验通过初步努力收集每取样面积四到六手套应用后立即开始时，以及在第二天早上当篷水分存在以确定手套的数量来传输最大农药残留。 </li></ol></li><li>下面样品采集完成后样品的采集，存储手套在玻璃罐暂时在冰或干冰在现场，并转移到冷冻3小时之内。 </li></ol></li></ol><img alt="图2" src="/files/ftp_upload/55182/55182fig2.jpg" /> 图2:手擦拭的方向。发展的目的是最大的独特股处理的植被面积联系，手套。两只手套需要避免采样&lt;415厘米当2区超载棉;然而，这应该在实验开始前除害剂和特定于站点的情况进行确认。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55182/55182fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="2"><li> 修改加利福尼亚辊 注:需要三个人。发现设备建筑规格在Full呃等。 9 <ol><li>使用辊（滚子14.5公斤，直径10厘米61厘米长铺上1.2厘米聚氨酯泡沫;处理:1.2米长）和帧（由91厘米长62厘米宽），一个棉片（&gt; 200线程数;70厘米99厘米长宽度）固定。 </li><li>有人士B和C的安全的棉片用夹子固定帧和固定取样面积与金属钉。 注:切之前的样品收集事件棉床单，以减少时间对于给定的集合。 </li><li>有人物A和B的安全的塑料（6毫米）的辊周围，以防止在整个样品污染。 </li><li>无添加往下压，有某甲滚动滚轮在棉布5次（一跌，后滚等于一次）。辊的转速应针对1.6±0.2公里/小时</li><li>有人物C从地下取出框架，并采取到附近的抽样表。 </li><li>虽然人C的保存帧，有某甲visua<html> LLY检查布草删除/土壤碎片用小镊子。 </li><li>有某甲取出棉布，并与某乙把它折叠，使得草坪与接触的一侧，折叠在一起，并存储在玻璃样本瓶。 <ol><li>暂时存储的玻璃罐中在该领域冰或干冰冷却器，和3小时内转移到冰柜以下样品收集完成。 </li></ol></li><li>下列样品的采集，丢弃围绕辊的塑料，清洁架，采样表和镊子用1:1氨:水，水和干重复该过程之前漂洗。 </li></ol></li></ol><img alt="图3" src="/files/ftp_upload/55182/55182fig3.jpg" /> 图3:球足球辊。不包括A到B，所有路口都粘有PVC不干胶。 C部分长度可以通过供应商，由于部分B和D的不同尺寸的变化e.com/files/ftp_upload/55182/55182fig3large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <ol start="3"><li> 足球球滚动 注:需要一个人。 <ol><li>用聚氯乙烯（PVC）管构成的足球辊架（40号;5厘米内直径）。 <ol><li>固定手柄（122厘米长; 图3A）到一个滑三通耦合器（ 图3B）。这是不是与PVC胶水固定，使更容易储存的唯一路口。 </li><li>放置在一个平面上（与对准辅助）的辊和固定的T形联接器的两侧滑动与7厘米管长度（ 图3C）90°弯管接头（ 图3D）。 </li><li>插入24厘米管道长度（ 图3E）到肘管接头的开口端</li><li>钻方头螺栓（0.6厘米直径7.5厘米长; 图3F）通过管道perpendicul的结束芳到手柄并平行于平的表面。 <ol><li>约18.4厘米方头螺栓之间的距离设置（可以跨品牌而异/足球的质量使用）。 </li><li>使轧（ 图3G）当足球不刺破文件/磨尖点关闭滞后螺栓用一只手的文件/磨床结束。 </li></ol></li></ol></li><li>释放足球的空气压力至24千帕。这虽然会underinflate球的时候从PVC滚筒松动，反而会增加压力，以约42千帕当球体积固定到滚筒后还原。 </li><li>通过触摸球端点（最大直径），以方头螺栓装入足球到滚筒。 注:要减少总体采样时间和潜在的污染，具有每采样1足球在给定的时间收集。 <ol><li>用手旋转球，以确保球滚是对称的。 </li><li>轻轻地按一下安装足球的顶部确保有足够的保护。 </li></ol></li><li>双环绕，与聚氯乙烯滚筒手柄，滚动（ 图3H），当它创建恒定草坪-吸附剂条接触对准球的圆周的吸附剂带（132厘米长5cm宽）。 <ol><li>固定吸附剂带与粘合带条（2.5厘米×2.5厘米）的球。 注:切之前的样品收集事件吸附剂条，以减少时间对于给定的集合。 </li></ol></li><li>超过3.7米的距离独特的治疗草坪的滚动球以恒定的速度（1.6±0.2公里/小时）。根据情节的尺寸，该距离可能需要并排侧辊来完成。这样做时，采取预防措施，联系不上植被尚未进行采样。 注:3.7米球滚动距离可能需要基于兴趣和草坪草系统的农药进行调整。这应该是事先确定通过preliminar试验启动ý努力收集不同距离应用后立即，以及在第二天早上的样品时篷水分存在，以确定最佳距离传输最大农药残留。 </li><li>继球滚动，取出吸附剂条和折叠，使得草坪与接触的一侧，折叠在一起，并存储在一个玻璃瓶。 <ol><li>暂时存储的玻璃罐中在该领域冰或干冰冷却器，和3小时内转移到冰柜以下样品收集完成。 </li></ol></li></ol></li></ol> 6.草坪植被收藏<ol><li>移除柱塞杯刀具内部保持取样设备从处理过的叶子（ 图4A）接触。 </li></ol><img alt="图4" src="/files/ftp_upload/55182/55182fig4.jpg" /> 图4:草坪核心种质。高尔夫球场杯刀是草坪/土壤收集一个强大的，相对便宜的设备。除去用于喷射用于转让农药残留样本的集合，以便植被不会无意中接触之前铁心内侧柱塞，这可以减少农药残留浓度。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55182/55182fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="2"><li> 嘉实代表质量的草坪核心。 <ol><li>使用刀杯勾勒的核心和切成5-7.5厘米土层深度去除帮助。 </li><li>在芯的边缘之外的对角线角从地面取出芯:用小刀切割（10〜15厘米叶片长度）。要特别注意防止与处理地上植被手/刀接触。 </li><li>平用剪刀底部土壤表面，这样它会在样品容器坐平。土壤深度应留出足够的空间，使得草坪草植物不接触样品容器的盖子。 注:如果天气允许，球场杯刀步骤（6.2.1协议），下午通常是建成后的树叶样品采集之前是干的，第二天早晨。 </li></ol></li></ol>

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Toxicol. 67 (6), 787-794 (2001).</li><li>Klonne, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+study+of+five+transferable+turf+residue+methods.">Comparative study of five transferable turf residue methods.</a> Bull. Environ. Contam. Toxicol. 67 (6), 771-779 (2001).</li><li>Rosenheck, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+a+standardized+sampling+technique+for+pesticide+transferable+turf+residues.">Determination of a standardized sampling technique for pesticide transferable turf residues.</a> Bull. Environ. Contam. Toxicol. 67 (6), 780-786 (2001).</li><li>Kim, D. S., Marshall, E. J. P., Brain, P., Caseley, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+crop+canopy+structure+on+herbicide+deposition+and+performance.">Effects of crop canopy structure on herbicide deposition and performance.</a> Weed Res. 51 (3), 310-320 (2011).</li><li>Jeffries, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+influencing+dislodgeable+2,4-D+plant+residues+from+hybrid+bermudagrass+(Cynodon+dactylon+L.+x+C.+transvaalensis.)+athletic+fields.">Factors influencing dislodgeable 2,4-D plant residues from hybrid bermudagrass (Cynodon dactylon L. x C. transvaalensis.) athletic fields.</a> PLoS One. 11 (2), 1-16 (2016).</li><li>Sears, M. K., Bowhey, C., Braun, H., Stephenson, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dislodgeable+residues+and+persistence+of+diazinon,+chloropyrifos+and+isofenphos+following+their+application+to+turfgrass.">Dislodgeable residues and persistence of diazinon, chloropyrifos and isofenphos following their application to turfgrass.</a> Pestic. Sci. 20 (3), 223-232 (1987).</li><li>Campillo, C., Prieto, M. H., Daza, C., Mo&#324;ino, M. J., Garc&#237;a, M. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+digital+images+to+characterize+canopy+coverage+and+light+interception+in+a+processing+tomato+crop.">Using digital images to characterize canopy coverage and light interception in a processing tomato crop.</a> HortSci. 43 (6), 1780-1786 (2008).</li></ol>

作者没有披露作出。

调节机构还没有确定具体的方法，从草坪量化转让农药残留。本研究利用支持基于现场和曝光过程特定的标准不同的方法，因为他们都对人类的风险评估工具。然而，它们都具有局限性，研究人员应该认识到之前它们的使用。最后，草种目前尚未转让的农药残留协议规定一个选址参数，这项研究建立在以前的努力表明其列入应予说明。 所描述的足球滚动的方法是在整个采样相对强劲，只需要一个人来完成。它也模仿了国际最流行的体育运动很普遍的情况，并有从手到口接触非饮食农药暴露很多问题。此外，这个概念可能与PE最小的修改就可以应用于sticide转移到与其他运动相关联的对象。但是，它是用于农药转移同等不太积极的方法，因此，不应只在风险评估利用。 的三种方法介绍，修改后的加州辊已在草坪残留转移就业的研究最为广泛。它是在整个采样最可靠的方法，并转移了类似的2,4-D的量作为评价最低的割草坪系统的其它方法，匍匐翦股颖。这表明，此方法应被用于高尔夫球场转让农药残留研究的果岭，以及其他可能密切割系统，如高尔夫球场球道/发球台和田径场。这种方法的局限性包括潜在的铺设草叶片跌修剪高度的增加和三个人的要求来完成。另外，样品C之间所需的辊和框架制备一个非常耗时，而且以前的研究已经表明农药转印波动在相对较短的时间尺度在一天内作为篷湿气消散14。这可能会限制在给定的定时（ 即减少处理）收集样品的量，或增加的数据的混杂因素应该样品在一个广泛的时间量被收集。研究人员应该意识到这一点，因为他们计划利用修改后的加州辊实验。 手抹处理植物植被与基于棉手套通常是用来衡量在果园和烟草工人转让农药残留由于与农业生产相关的手到植被的接触频率高的方法。虽然这种方法已在草皮草系统被利用以下时，提供了优越的方法在修剪高度来量化草坪系统转让残基通常与commercia相关联L /住宅草坪和公园。此外，用手处理的草坪草接触是两个非职业性和职业危险评估常见，如草坪系统被用于各种社会的目的。三种评价的方法，手工擦拭横跨采样所述至少再现的方法，该方法可能需要额外的措施（复制，培训等 ），以产生决定性的结果。 虽然评价方法在它们的执行广泛地变化，每个协议中的关键步骤概念上重叠。以恒定的速度和压力采样是极为重要的，以产生再现的数据，因为这些影响吸附剂材料农药传输。保持恒定的速度，需要在所有三种方法取样，而压力为手仅擦拭关注的点。取样不应该把修改后的加利福尼亚州或足球辊额外的压力，同时用手擦拭是一种方法，需要substanTiAl基合金的初步努力保持内的一致性，和整个取样。这是最大的限制转让的残留研究只手抹依托，和今后的研究应该确定一个更客观的方法，提供了其独特的树冠穿透属性，同时最低限度地铺设草了。 收集草坪草植物的目的是，除了最初由占应用和随后的样本采集定时之间耗散施加的农药的量提供一个参考点。此外，在植被量化农药残留未检测时转移样品中出现增强了解释。基本上，它使研究人员，以确定是否是因为农药残留物吸附在/植被没有发生转移，使之成为不可转让的，或者残留不再检测中/植被。

草坪草在美国本土（美国）超过16300000公顷增长-超过粮食灌溉的玉米联合区域[（ 玉米 L.）2500000公顷]，大豆[（ 大豆 L.）2100000公顷]和棉花[（ 陆地棉 ）0900000公顷] -和公众提供土地使用用途，包括田径场，商业/住宅草坪，高尔夫球场和公园1，2。他们提供了许多积极的社会属性，包括防尘，散热，娱乐的表面和土壤稳定。然而，可能会出现害虫侵害这需要使用农药（多个）的草皮维持在可接受的水平3。 在建立草坪系统截获植物冠层喷洒农药。人类接触杀虫剂通过各种途径是可能的，包括治疗已经转移getation直接到人，以及通过间接的途径，如转移到维修设备，宠物和游乐项目4,5。为了解决这样的担心，人类接触杀虫剂风险评估必须农药登记和重新登记之前在美国进行估算暴露于污染环境介质6不良健康影响的性质和概率。在目前由美国环保局采用的职业和住宅暴露试验的指导方针，叶面转让残留损耗测试（OPPTS 875.2100）会传导到可以通过各种方法转移到人的皮肤/服装或吸入7,8处理过的表面上量化农药残留。 以往的研究努力已经比较了几种叶面转让农药残留方法inclu丁加州辊（移动已知质量超过棉布的辊），拖动雪橇（拉已知质量的固体对象，具有连接到它一块布），聚氨酯泡沫辊（移动已知质量的辊覆盖有聚氨酯泡沫）和鞋改组（附着棉花纱布到鞋），这是在一个已知的区域内的所有进行农药处理的草坪草9，10，11，12。在上述方法中，根据加州辊的方法提供了最可重复的方法来量化叶面转让的农药残留;然而，同等更积极的方法，比如鞋洗牌可以传递更多的农药残留，其中也有在风险评估工具。手擦拭方法提供一个相对增强的联系独特的治疗草坪植被表面的能力。这种方法提供了更多的nondieta适用的数据RY农药吞食，手到口的接触与此的暴露途径相关的通用流程。 草坪篷动态物种和使用站点之间的差别很大。种在生长习性和季节通常会发生变化，以及刀片的质地和密度，这会影响农药喷雾拦截和生理过程3中 ，13。管理投入，才能使用网站之间有很大的不同，并根据现场具体期望使用的站点内。例如，狗牙根（ 狗牙根属）是在0.3至0.4厘米，以及一个灌溉的效用地面覆盖用在适于气候作为高尔夫球场球洞表面这是典型的灌溉和割每周（收集剪辑）&gt; 5倍可割的&lt;每周1次（剪报返回）在1.5至6.3厘米此前有研究表明转让叶面残留农药可以使用一个站点内的物种之间变化，并通过灌溉和割草做法14,15的影响。最终，草坪系统之间的变异抑制量化转让叶面残留农药的通用方法的实现。因此，方法选择，以优化人类的风险评估应该包括除害剂，流程 - ，现场和物种的具体标准。本研究的目的是表征用于量化转让叶面农药残留的各种方法，并突出显示应选择用于一个给定的实验的方法时，必须考虑的条件。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>General</td>
			<td>---</td>
			<td>---</td>
		</tr>
		<tr>
			<td>Nitrile gloves</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Coolers</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Turf paint</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Field plot measuring equipment</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Protective foot apparel</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Whatman 3 MM Chr Chromatography Paper</td>
			<td>Fisher Scientific</td>
			<td>05-716-3E</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
		</tr>
		<tr>
			<td>Ball roll</td>
			<td>---</td>
			<td>---</td>
		</tr>
		<tr>
			<td>PVC pipe (5 cm inner diameter)</td>
			<td>Home Depot</td>
			<td>531137</td>
		</tr>
		<tr>
			<td>Hacksaw for PVC cutting</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>90 degree elbow (5 cm inner diameter)</td>
			<td>Home Depot</td>
			<td>RCE-2000-S</td>
		</tr>
		<tr>
			<td>Tee coupler (5 cm inner diameter)</td>
			<td>Home Depot</td>
			<td>PVC024001600HD</td>
		</tr>
		<tr>
			<td>PVC adhesive</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Lag bolt (0.6 cm diameter by 7 cm length)</td>
			<td>Home Depot</td>
			<td>801366</td>
		</tr>
		<tr>
			<td>Size 4 soccer ball</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Pressure gauge</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Hand air pump</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Fabric scissors</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Scott Rags-In-A-Box</td>
			<td>Uline</td>
			<td>S-12809</td>
		</tr>
		<tr>
			<td>Adhesive tape</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>8 oz sealable glass jar</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
		</tr>
		<tr>
			<td>Hand wipe</td>
			<td>---</td>
			<td>---</td>
		</tr>
		<tr>
			<td>100% cotton heavyweight inspection gloves</td>
			<td>Uline</td>
			<td>S-19284</td>
		</tr>
		<tr>
			<td>Digital camera</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>ImageJ software</td>
			<td>National Institutes of Health</td>
			<td>https://imagej.nih.gov/ij/</td>
		</tr>
		<tr>
			<td>Digital scale that measures up to 400 g</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Stopwatch</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Plastic bucket (23 cm diameter)</td>
			<td>Home Depot</td>
			<td>209313</td>
		</tr>
		<tr>
			<td>16 oz sealable glass jar</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
		</tr>
		<tr>
			<td>Modifed California roller</td>
			<td>---</td>
			<td>---</td>
		</tr>
		<tr>
			<td>Metal conduit (1.25 cm diameter by 1.8 m length)</td>
			<td>Home Depot</td>
			<td>101543</td>
		</tr>
		<tr>
			<td>PVC pipe (10 cm inner diameter)</td>
			<td>Home Depot</td>
			<td>531103</td>
		</tr>
		<tr>
			<td>Sand + reebar to bring roller to 14.5 kg</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>PVC plug</td>
			<td>Home Depot</td>
			<td>33403D</td>
		</tr>
		<tr>
			<td>Polyurethane foam sheet (1.25 cm thick) to cover PVC pipe</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>6 mm painter&#39;s plastic</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Plexiglass sheet (107 cm length by 76 cm width by 0.6 cm thick)</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Toggle clamps</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Metal nails (10 to 15 cm length) to secure frame to ground</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>100% cotton sheets (&#62; 200 threadcount)</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Tweezers</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>32 oz sealable glass jar</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
		</tr>
		<tr>
			<td>Turfgrass vegetation core collection</td>
			<td>---</td>
			<td>---</td>
		</tr>
		<tr>
			<td>Lever action golf course cup cutter</td>
			<td>Par Aide Product Company</td>
			<td>1001-1</td>
		</tr>
		<tr>
			<td>Knife</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
		<tr>
			<td>Fiskars Herb Scissors</td>
			<td>Home Depot</td>
			<td>96086966J</td>
		</tr>
		<tr>
			<td>Turf plug plastic container</td>
			<td>Any</td>
			<td>NA</td>
		</tr>
	</tbody>
</table>

protocols for quantifying transferable pesticide residues in turfgrass systems

在建立了草坪系统植物冠层可以拦截喷洒杀虫剂，这可以通过各种途径被转移到人类可察觉量。为此，由美国国家环境保护局（USEPA）需要注册和重新注册转让的农药残留试验。尽管需要这样的实验，需要有限的特异性有关实验方法。在三个处理草种用来评估农药转移到人类，包括手用棉手套擦实验方法，修改加利福尼亚辊（移动已知质量比棉布的辊）和足球球滚动（球包裹着吸附剂条）（匍匐翦股颖，杂种狗牙根，分别维持在0.4，5和9厘米高羊茅）给出。改性加利福尼亚辊是最广泛使用的方法来日期，是最适合于采用在低割草高度由于其再现性和大取样区。足球滚动是一个不太积极的传输方式;但是，它模仿了国际最流行的体育运动很普遍的情况，并有从手到口接触非饮食农药暴露很多问题。此外，这种方法可能具有有限的修改其他运动活动进行调节。手抹是在更高的修剪高度转让农药，基于辊的方法才能奠定在叶片的最佳方法;然而，这是由于更多的可变取样压力比较主观。横跨草坪草这些方法的效用提出，并额外考虑进行转让的农药残留研究草坪系统进行了讨论。

在转移单农药残留方法中的前期研究工作的单一草坪系统内比较转让农药残留的方法，以及草坪系统的基础上，实地研究（发起的2016年5月24日在北卡罗来纳州罗利市，美国）进行跨比较方法草坪系统。总之，2,4-D，使用常用于草坪系统一个阔叶除草剂，从三个草坪草物种（匍匐翦股颖转移， 匍匐翦股颖 L。;杂交狗牙根， 狗牙根属点¯xC. transvaalensis伯特-戴维;高羊茅， 黑麦草茅 [茅。]通过三种方法SJ Darbyshire）（手擦，改性加利福尼亚辊或足球辊）被应用程序和1小时干燥期之后立即进行定量，以及1和3的DAT。匍匐翦股颖修剪高度模拟高尔夫球场果岭0.4厘米，而目前混合型狗牙根和tal升羊茅分别维持在5和9厘米，，这是典型的商业/住宅草坪和公园。研究领域没有割完并按照广播2,4-D喷涂（1公斤酸当量公顷-1）降雨过程覆盖。 相比以往的报告处理的草坪草评估转移的2,4-D，从所呈现的研究的数据表明条件为在所有的方法传递是有利的。平均方法，2,4-D立即转移申请后排名匍匐翦股颖（应用于21％）&gt;混合型狗牙根（16.4％）=高羊茅（15.1％），这与整个系统的郁闭度的趋势（ 见表1对齐; 图5）。平均草坪草，2,4-D转换后的应用程序排名的手擦拭（21.2％应用）&gt;修改加利福尼亚辊（16.8％）=足球球滚动（14.4％），这与以前的努力展示同意ING手/鞋抽样可以相对于其他传输技术，包括拖拽和基于辊的方法10，11传输更多的农药残留。的1 H干燥期间导致整个草坪草和改性加利福尼亚辊和手擦拭方法2至4倍的降低在转让的2,4-D残留，而转移减少从足球辊36倍。这种下降与杰弗里斯等人的同意。 14，谁报告经由足球辊2,4-D的传输从11.2％下降一个1小时干燥期之后施加于杂交狗牙根施用后立即到0.3％。这些数据强调应用效果样本采集的时间和方法，在从草坪量化转让的残留物。足球辊是在草坪系统相对特定的过程，虽然它提供了一个运动场人类接触相关的信息，它可能无法适当送是<html> olely利用一般人体暴露风险评估的其他方法。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td fo:text-align="center"></td><td fo:text-align="center" style="text-align: center;">匍匐翦股颖</td><td fo:text-align="center" style="text-align: center;">混合狗牙根</td><td fo:text-align="center" style="text-align: center;">高羊茅</td><td fo:text-align="center" style="text-align: center;">球滚动</td><td fo:text-align="center" style="text-align: center;">国防部。卡尔。滚筒</td><td fo:text-align="center" style="text-align: center;">手擦</td></tr><tr><td fo:text-align="center"></td><td colspan="6" fo:text-align="center" style="text-align: center;"> _________________________％用于2,4-D的脱落_________________________ </td></tr><tr><td fo:text-align="center"> DAT 0 - 0 H </td>对齐:中心;"&gt; 21 </td><td fo:text-align="center" style="text-align: center;"> 16.4 </td><td fo:text-align="center" style="text-align: center;"> 15.1 </td><td fo:text-align="center" style="text-align: center;"> 14.4 </td><td fo:text-align="center" style="text-align: center;"> 16.8 </td><td fo:text-align="center" style="text-align: center;"> 21.2 </td></tr><tr><td fo:text-align="center"> LSD 0.05 </td><td colspan="3" fo:text-align="center" style="text-align: center;"> _______________ _______________ 2.8 </td><td colspan="3" fo:text-align="center" style="text-align: center;"> _______________ _______________ 2.8 </td></tr><tr><td fo:text-align="center"> 0的DAT - 1小时</td><td fo:text-align="center" style="text-align: center;">五</td><td fo:text-align="center" style="text-align: center;"> 6.8 </td><td fo:text-align="center" style="text-align: center;"> 4.9 </td><td fo:text-align="center" style="text-align: center;"> 0.4 </td><td fo:text-alIGN ="中心"的风格="文本对齐:中心;"&gt; 7.7 </td><td fo:text-align="center" style="text-align: center;"> 8.5 </td></tr><tr><td fo:text-align="center"> LSD 0.05 </td><td colspan="3" fo:text-align="center" style="text-align: center;"> _______________ _______________ 1.0 </td><td colspan="3" fo:text-align="center" style="text-align: center;"> _______________ _______________ 1.0 </td></tr></tbody></table> 表1:从田间小区转换2,4-D应用程序的一天。草种和报告为初次申请率％的转让2,4-D数据采样的方法主要作用。样品收集发生后立即应用程序和一个1小时的干燥时间后。 <img alt="图5" src="/files/ftp_upload/55182/55182fig5.jpg" /> 图5:草坪系统评价。 Turfgras小号郁闭度和高度可以通过系统而异。在所提出的研究，密度（从高到低）排名匍匐翦股颖（A）&gt;混合型狗牙根（B）&gt;高羊茅（C）;而高度（最高到最低）排名高羊茅&gt;混合型狗牙根&gt;匍匐翦股颖。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55182/55182fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 从1和3 DAT数据表明，样品采集方法不从治疗植被同样横跨草坪草，这是假设，由于不同树冠动态转移2,4-D。内的1 DAT，从杂种狗牙根2,4-D转换（17.3至31.2的施加％）的样品采集方法比匍匐翦股颖（10.6至16.2％）和高羊茅（8.1至20.9％）时，这很可能是部分原因是由于不同的树冠达因AMICS和跨物种除草剂生理作用（ 表2）。阐明这发生超出了本实验的范围;然而，它突出展示草种选择了转让农药残留研究的重要性。转换的2,4-D没有在1或3的DAT，这是最好的质感，最低割草坪评价上匍匐翦股颖方法之间变化。这允许在三个评价方法相对一致的吸着剂材料处理过的植物接触。横跨在杂交狗牙根和高羊茅方法2,4-D转换而变化，以产生最大的转印手擦拭。杂种狗牙根和高羊茅被粗糙纹理比匍匐翦股颖，并在更高的高度（5和9厘米，分别），以突出的铺设植被以上（ 图6）的基于轧制的方法的固有限制被割。当这种情况发生时，吸着剂材料处理过的植物接触可以降低，因此，低估转让残基。 <table fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td fo:text-align="center" style="text-align: center;"></td><td colspan="4" fo:text-align="center" style="text-align: center;"> ____________________ 1 DAT - 7:00 EST ____________________ </td></tr><tr><td fo:text-align="center" style="text-align: center;">草坪</td><td fo:text-align="center" style="text-align: center;">球滚动</td><td fo:text-align="center" style="text-align: center;">国防部。卡尔。滚筒</td><td fo:text-align="center" style="text-align: center;">手擦</td><td fo:text-align="center" style="text-align: center;"> LSD 0.05 </td></tr><tr><td fo:text-align="center" style="text-align: center;"></td><td colspan="4" fo:text-align="center" style="text-align: center;"> ____________％用于2,4-D的脱落____________ </td></tr><tr><td fo:text-align="center" style="text-align: center;">匍匐翦股颖</td><td fo:text-align="center" style="text-align: center;"> 10.6 </td><td fo:text-align="center" style="text-align: center;"> 13.6 </td><td fo:text-align="center" style="text-align: center;"> 16.2 </td><td fo:text-align="center" style="text-align: center;"> NS </td></tr><tr><td fo:text-align="center" style="text-align: center;">混合狗牙根</td><td fo:text-align="center" style="text-align: center;"> 17.3 </td><td fo:text-align="center" style="text-align: center;"> 20.9 </td><td fo:text-align="center" style="text-align: center;"> 31.2 </td><td fo:text-align="center" style="text-align: center;"> 2.2 </td></tr><tr><td fo:text-align="center" style="text-align: center;">高羊茅</td><td fo:text-align="center" style="text-align: center;"> 8.1 </td><td fo:text-align="center" style="text-align: center;"> 9.1 </td><td fo:text-align="center" style="text-align: center;">20.9 </td><td fo:text-align="center" style="text-align: center;"> 2.7 </td></tr><tr><td fo:text-align="center" style="text-align: center;"> LSD 0.05 </td><td fo:text-align="center" style="text-align: center;"> 3.2 </td><td fo:text-align="center" style="text-align: center;"> 3.2 </td><td fo:text-align="center" style="text-align: center;"> 5.2 </td><td fo:text-align="center" style="text-align: center;"></td></tr><tr><td fo:text-align="center" style="text-align: center;"></td><td colspan="4" fo:text-align="center" style="text-align: center;"> ____________________ 3 DAT - 7:00 EST ____________________ </td></tr><tr><td fo:text-align="center" style="text-align: center;">草坪</td><td fo:text-align="center" style="text-align: center;">球滚动</td><td fo:text-align="center" style="text-align: center;">国防部。卡尔。滚筒</td><td fo:text-align="center" style="text-align: center;">手擦</td><td fo:text-align="center" stylE ="文本对齐:中心;"&gt; LSD 0.05 </td></tr><tr><td fo:text-align="center" style="text-align: center;"></td><td colspan="4" fo:text-align="center" style="text-align: center;"> ____________％用于2,4-D的脱落____________ </td></tr><tr><td fo:text-align="center" style="text-align: center;">匍匐翦股颖</td><td fo:text-align="center" style="text-align: center;"> 1.9 </td><td fo:text-align="center" style="text-align: center;"> 2.8 </td><td fo:text-align="center" style="text-align: center;"> 3.1 </td><td fo:text-align="center" style="text-align: center;"> NS </td></tr><tr><td fo:text-align="center" style="text-align: center;">混合狗牙根</td><td fo:text-align="center" style="text-align: center;"> 1.9 </td><td fo:text-align="center" style="text-align: center;"> 4.8 </td><td fo:text-align="center" style="text-align: center;"> 7.6 </td><td fo:text-align="center" style="text-align: center;"> 2.1 </td></tr><tr&gt; <td fo:text-align="center" style="text-align: center;">高羊茅</td><td fo:text-align="center" style="text-align: center;"> 1.8 </td><td fo:text-align="center" style="text-align: center;"> 2 </td><td fo:text-align="center" style="text-align: center;"> 6.8 </td><td fo:text-align="center" style="text-align: center;"> 2.6 </td></tr><tr><td fo:text-align="center" style="text-align: center;"> LSD 0.05 </td><td fo:text-align="center" style="text-align: center;"> NS </td><td fo:text-align="center" style="text-align: center;"> 0.9 </td><td fo:text-align="center" style="text-align: center;"> 3.4 </td><td fo:text-align="center" style="text-align: center;"></td></tr></tbody></table> 表2:从田间小区1和应用后3天转换的2,4-D。关于报告作为初始APPLICA的百分之转让的2,4-D数据草坪草物种逐采样方法相互作用化率。样品收集发生在7:00:00东部标准时间。 <img alt="图6" src="/files/ftp_upload/55182/55182fig6.jpg" /> 图6:以下修改加州滚子采样高羊茅檐篷。这一转让农药残留抽样方法固有的限制越来越多的草叶潜力躺下的树冠高度增加。当这种情况发生时，背面表面不被棉吸附剂片接触，从而，有可能低估转让残基。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55182/55182fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Protocols for Quantifying Transferable Pesticide Residues in Turfgrass Systems at JoVE.com

Protocols for Quantifying Transferable Pesticide Residues in Turfgrass Systems

在草坪系统转让的农药残留试验是人类风险评估的组成部分。测量转让残实验方法应调整到感兴趣和草坪草系统动力学的人工交互。三个转让农药残留协议提出并在三种草坪草系统的适用性进行了讨论。

协议在草坪系统量化转换农药残留

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

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Nanyang Technological University

Research

JoVE Journal

Environment

7.2K 次观看.  North Carolina State University. 在草坪系统转让的农药残留试验是人类风险评估的组成部分。测量转让残实验方法应调整到感兴趣和草坪草系统动力学的人工交互。三个转让农药残留协议提出并在三种草坪草系统的适用性进行了讨论。 

视频: Protocols for Quantifying Transferable Pesticide Residues in Turfgrass Systems

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is not an appropriate measure for its availability; bioavailability rather depends on the dynamic interplay of potential mass transfer (flux) of a compound to a microbial cell and the capacity of the latter to degrade the compound. In water-unsaturated parts of the soil, mycelia have been shown to overcome bioavailability limitations by actively transporting and mobilizing organic compounds over the range of centimeters. Whereas the extent of mycelia-based transport can be quantified easily by chemical means, verification of the contaminant-bioavailability to bacterial cells requires a biological method. Addressing this constraint, we chose the PAH fluorene (FLU) as a model compound and developed a water unsaturated model microcosm linking a spatially separated FLU point source and the FLU degrading bioreporter bacterium Burkholderia sartisoli RP037-mChe by a mycelial network of Pythium ultimum. Since the bioreporter expresses eGFP in response of the PAH flux to the cell, bacterial FLU exposure and degradation could be monitored directly in the microcosms via confocal laser scanning microscopy (CLSM). CLSM and image analyses revealed a significant increase of the eGFP expression in the presence of P. ultimum compared to controls without mycelia or FLU thus indicating FLU bioavailability to bacteria after mycelia-mediated transport. CLSM results were supported by chemical analyses in identical microcosms. The developed microcosm proved suitable to investigate contaminant bioavailability and to concomitantly visualize the involved bacteria-mycelial interactions.

A whole cell bioreporter assay with Burkholderia sartisoli RP037-mChe was developed to detect fractions of an organic contaminant (i.e., fluorene) available for bacterial degradation after active transport by mycelia bridging air-filled pores in a water unsaturated model system.

全细胞Bioreporter方法​​评估交通有机污染物的生物利用度和水饱和系统

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is ...

科研

环境科学

A New Application of the Electrical Penetration Graph (EPG) for Acquiring and Measuring Electrical Signals in Phloem Sieve Elements

Electrophysiological properties of cells are often studied in vitro, after dissociating them from their native environments. However, the study of electrical transmission between distant cells in an organism requires in vivo, artifact-free recordings of cells embedded within their native environment. The transmission of electrical signals from wounded to unwounded areas in a plant has since long piqued the interest of botanists. The phloem, the living part of the plant vasculature that is spread throughout the plant, has been postulated as a major tissue in electrical transmission in plants. The lack of suitable electrophysiological methods poses many challenges for the study of the electrical properties of the phloem cells in vivo. Here we present a novel approach for intracellular electrophysiology of sieve elements (SEs) that uses living aphids, or other phloem-feeding hemipteran insects, integrated in the electrical penetration graph (EPG) circuit. The versatility, robustness, and accuracy of this method made it possible to record and study in detail the wound-induced electrical signals in SEs of central veins of the model plant Arabidopsis thaliana1. Here we show that EPG-electrodes can be easily implemented for intracellular electrophysiological recordings of SEs in marginal veins, as well as to study the capacity of SEs to respond with electrical signals to several external stimuli. The EPG approach applied to intracellular electrophysiology of SEs can be implemented to a wide variety of plant species, in a large number of plant/insect combinations, and for many research aims.

A New Application of the Electrical Penetration Graph EPG for Acquiring and Measuring Electrical Signals in Phloem Sieve Elements

Electrical Penetration Graph (EPG) is a well-established technique for studying the feeding behavior of stylet-bearing insects. Here we show a new application of EPG as a non-invasive tool for the acquisition of intracellular electrophysiology recordings of sieve elements (SEs), the cells that form the phloem vasculature in plants.

在电气贯穿图（EPG）的韧皮部筛分子获取和测量电信号中的新应用

Electrophysiological properties of cells are often studied in vitro, after dissociating them from their native environments. However, the study of ...

Protocols for Robust Herbicide Resistance Testing in Different Weed Species

Robust protocols to test putative herbicide resistant weed populations at whole plant level are essential to confirm the resistance status. The presented protocols, based on whole-plant bioassays performed in a greenhouse, can be readily adapted to a wide range of weed species and herbicides through appropriate variants. Seed samples from plants that survived a field herbicide treatment are collected and stored dry at low temperature until used. Germination methods differ according to weed species and seed dormancy type. Seedlings at similar growth stage are transplanted and maintained in the greenhouse under appropriate conditions until plants have reached the right growth stage for herbicide treatment. Accuracy is required to prepare the herbicide solution to avoid unverifiable mistakes. Other critical steps such as the application volume and spray speed are also evaluated. The advantages of this protocol, compared to others based on whole plant bioassays using one herbicide dose, are related to the higher reliability and the possibility of inferring the resistance level. Quicker and less expensive in vivo or in vitro diagnostic screening tests have been proposed (Petri dish bioassays, spectrophotometric tests), but they provide only qualitative information and their widespread use is hindered by the laborious set-up that some species may require. For routine resistance testing, the proposed whole plant bioassay can be applied at only one herbicide dose, so reducing the costs.

A robust and flexible approach to confirm herbicide resistance in weed populations is presented. This protocol allows the herbicide resistance levels to be inferred and applied to a wide range of weed species and herbicides with minor adaptations.

协议的鲁棒抗除草剂试验在不同杂草

Robust protocols to test putative herbicide resistant weed populations at whole plant level are essential to confirm the resistance status. The presented ...

LC-MS Analysis of Human Platelets as a Platform for Studying Mitochondrial Metabolism

Perturbed mitochondrial metabolism has received renewed interest as playing a causative role in a range of diseases. Probing alterations to metabolic pathways requires a model in which external factors can be well controlled, allowing for reproducible and meaningful results. Many studies employ transformed cellular models for these purposes; however, metabolic reprogramming that occurs in many cancer cell lines may introduce confounding variables. For this reason primary cells are desirable, though attaining adequate biomass for metabolic studies can be challenging. Here we show that human platelets can be utilized as a platform to carry out metabolic studies in combination with liquid chromatography-tandem mass spectrometry analysis. This approach is amenable to relative quantification and isotopic labeling to probe the activity of specific metabolic pathways. Availability of platelets from individual donors or from blood banks makes this model system applicable to clinical studies and feasible to scale up. Here we utilize isolated platelets to confirm previously identified compensatory metabolic shifts in response to the complex I inhibitor rotenone. More specifically, a decrease in glycolysis is accompanied by an increase in fatty acid oxidation to maintain acetyl-CoA levels. Our results show that platelets can be used as an easily accessible and medically relevant model to probe the effects of xenobiotics on cellular metabolism.

Here we show isolated human platelets can be used as an accessible ex vivo model to study metabolic adaptations in response to the complex I inhibitor rotenone. This approach employs isotopic tracing and relative quantification by liquid chromatography-mass spectrometry and can be applied to a variety of study designs.

人类血小板的LC-MS分析作为研究线粒体代谢的平台

Perturbed mitochondrial metabolism has received renewed interest as playing a causative role in a range of diseases. Probing alterations to metabolic ...

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 ...

Implementation of Portable Emissions Measurement Systems (PEMS) for the Real-driving Emissions (RDE) Regulation in Europe

Vehicles are tested in controlled and relatively narrow laboratory conditions to determine their official emission values and reference fuel consumption. However, on the road, ambient and driving conditions can vary over a wide range, sometimes causing emissions to be higher than those measured in the laboratory. For this reason, the European Commission has developed a complementary Real-Driving Emissions (RDE) test procedure using the Portable Emissions Measurement Systems (PEMS) to verify gaseous pollutant and particle number emissions during a wide range of normal operating conditions on the road. This paper presents the newly-adopted RDE test procedure, differentiating six steps: 1) vehicle selection, 2) vehicle preparation, 3) trip design, 4) trip execution, 5) trip verification, and 6) calculation of emissions. Of these steps, vehicle preparation and trip execution are described in greater detail. Examples of trip verification and the calculations of emissions are given.

Implementation of Portable Emissions Measurement Systems PEMS for the Real-driving Emissions RDE Regulation in Europe

The European Commission has developed a Real-Driving Emissions (RDE) test procedure to verify pollutant emissions during real-world vehicle operation using the Portable Emissions Measurement Systems (PEMS). This paper presents the experimental procedures required by the newly-adopted RDE test.

便携式排放测量系统（PEMS）在欧洲的真实驾驶排放（RDE）规例的执行情况

Vehicles are tested in controlled and relatively narrow laboratory conditions to determine their official emission values and reference fuel consumption. ...

Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may be treated with simple systems designed to promote sorption of contaminants to vegetation and soils and promote infiltration. Two example systems are described: a bioswale treatment system for urban stormwater treatment, and a vegetated drainage ditch for treating agriculture irrigation runoff. Both have similar attributes that reduce contaminant loading in runoff: vegetation that results in sorption of the contaminants to the soil and plant surfaces, and water infiltration. These systems may also include the integration of granulated activated carbon as a polishing step to remove residual contaminants. Implementation of these systems in agriculture and urban watersheds requires system monitoring to verify treatment efficacy. This includes chemical monitoring for specific contaminants responsible for toxicity. The current paper emphasizes monitoring of current use pesticides since these are responsible for surface water toxicity to aquatic invertebrates.

This article summarizes the design attributes and the effectiveness of treatment systems that treat urban stormwater and agriculture irrigation runoff to remove pesticides and other contaminants associated with aquatic toxicity.

用于去除与农业和城市径流中地表水毒性有关的污染物的植被处理系统

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may ...

RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop cultivars against environmental stresses. An experimental protocol is presented for RGB and hyperspectral imaging of root systems. The approach uses rhizoboxes where plants grow in natural soil over a longer time to observe fully developed root systems. Experimental settings are exemplified for assessing rhizobox plants under water stress and studying the role of roots. An RGB imaging setup is described for cheap and quick quantification of root development over time. Hyperspectral imaging improves root segmentation from the soil background compared to RGB color based thresholding. The particular strength of hyperspectral imaging is the acquisition of chemometric information on the root-soil system for functional understanding. This is demonstrated with high resolution water content mapping. Spectral imaging however is more complex in image acquisition, processing and analysis compared to the RGB approach. A combination of both methods can optimize a comprehensive assessment of the root system. Application examples integrating root and aboveground traits are given for the context of plant phenotyping and plant physiological research. Further improvement of root imaging can be obtained by optimizing RGB image quality with better illumination using different light sources and by extension of image analysis methods to infer on root zone properties from spectral data.

An experimental protocol is presented for assessment of soil grown plant root systems with RGB and hyperspectral imaging. Combination of RGB image time series with chemometric information from hyperspectral scans optimizes insights into plant root dynamics.

RGB 和光谱根成像植物表型和生理研究: 实验装置和成像协议

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop ...

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

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

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

利用水下立体视频工具量化鱼类密度的新方法研究进展

The use of video camera systems in ecological studies of fish continues to gain traction as a viable, non-extractive method of measuring fish lengths and ...

Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

Beneficial plant-microbe interactions offer a sustainable biological solution with the potential to boost low-input food and bioenergy production. A better mechanistic understanding of these complex plant-microbe interactions will be crucial to improving plant production as well as performing basic ecological studies investigating plant-soil-microbe interactions. Here, a detailed description for ecosystem fabrication is presented, using widely available 3D printing technologies, to create controlled laboratory habitats (EcoFABs) for mechanistic studies of plant-microbe interactions within specific environmental conditions. Two sizes of EcoFABs are described that are suited for the investigation of microbial interactions with various plant species, including Arabidopsis thaliana, Brachypodium distachyon, and Panicum virgatum. These flow-through devices allow for controlled manipulation and sampling of root microbiomes, root chemistry as well as imaging of root morphology and microbial localization. This protocol includes the details for maintaining sterile conditions inside EcoFABs and mounting independent LED light systems onto EcoFABs. Detailed methods for addition of different forms of media, including soils, sand, and liquid growth media coupled to the characterization of these systems using imaging and metabolomics are described. Together, these systems enable dynamic and detailed investigation of plant and plant-microbial consortia including the manipulation of microbiome composition (including mutants), the monitoring of plant growth, root morphology, exudate composition, and microbial localization under controlled environmental conditions. We anticipate that these detailed protocols will serve as an important starting point for other researchers, ideally helping create standardized experimental systems for investigating plant-microbe interactions.

Ecosystem Fabrication EcoFAB Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

This article describes detailed protocols for ecosystem fabrication of devices (EcoFABs) that enable the studies of plants and plant-microbe interactions in highly controlled laboratory conditions.

用于研究植物-微生物相互作用的实验室生态系统制造 (EcoFAB) 协议

Beneficial plant-microbe interactions offer a sustainable biological solution with the potential to boost low-input food and bioenergy production. A ...