We thank Dr. Roger Hopper and the Wyoming Department of Agriculture, Analytical Services Lab for the mass spectroscopy analysis of our samples. We would like to express our gratitude to the University of Wyoming, School of Pharmacy for allowing us to video this protocol in their laboratories. We would also like to thank the Theodore O. and Dorothy S. King Endowed Professorship Agreement for their graduate assistantship (SC), the University of Wyoming for the Graduate Assistantship support (JRS), and the Science Posse (NSF GK-12 Project # 084129) for the teaching fellowship (JRS). We would also like to thank Uranium One for allowing us to obtain samples and assisting us with questions. This work was supported by the School of Energy Resources, University of Wyoming.

所有样本收集在怀俄明州铀ISR设施铀液体处理建设。 1.生产渗水（PBW） <ol><li>收集两种类型的水样从ISR铀设施：PBW和反渗透（RO）水。从离子交换处理后的监测抽头但反渗透净化之前收集PBW。收集的RO样品后PBW通过反渗透处理去污。 注：浸滤剂输送来自多个井场的铀处理液体的建筑物，在那里被收集在柱和离子交换制得的管道。大约经过离子交换的浸滤剂的1-3％被从电路移除，称为生产排出水（PBW）。 PBW被重新用在挖掘过程或净化/脱钙用RO过滤。 </li><li>收集在高密度聚乙烯水样（HDPE）瓶，根据零头部空间为样品采集和环境质量的怀俄明部门（WYDEQ）23分析标准作业程序。 </li><li>测量温度和pH现场及运输样品在冰上，让他们冷静。 </li><li>商店PBW在4℃。保持PBW液冷却，直至浓缩鹰的最低基本培养基（EMEM-10X），此前有媒体在制备过程中加入的指示在以下协议。 注：PBW是氧化的溶液，将沉淀如果允许冻结或温热至室温。稀释后的PBW溶液足够稀该应用之前它不会沉淀当加热到37℃至细胞和孵育期间。 </li></ol> 2.制备氧化铜纳米粒子（的CuO-NPS） <ol><li>结合含有250mM毫升的0.2M的CuCl 2•2H 2 O，加入250毫升的0.4M的氢氧化钠（NaOH），和5克聚乙二醇纯乙醇溶液（PEG）在一个圆底烧瓶中6毫米硼硅酸盐玻璃球。 </li><li>放置在修改后的微波炉的溶液，并允许它在20％的功率（的6秒的时间间隔，24秒关）在回流下反应，在环境空气压力10分钟。 </li><li>冷却该溶液到室温（20℃），然后倾入50ml锥形管中，留下的玻璃球。 </li><li>离心机在1000×g离心30分钟，滗析该溶液在50ml锥形管中，然后洗的CuO纳米粒用300毫升热水（60-65℃），100毫升乙醇中的序列，和100毫升丙酮。 </li><li>干燥的CuO纳米粒在50ml锥形管中室温（20℃）。 </li><li>刮氧化铜纳米粒他们的管研钵中。覆盖的CuO纳米粒用锡箔和加热的CuO纳米粒至110℃的炉中以除去残留的液体。结合氧化铜纳米粒为一个批次，重量CuO的纳米粒。 注：氧化铜纳米粒和CuO-NP处理PBW的制备是在水中进行的Qual生态系统科学与管理，怀俄明大学的实验室性。的CuO-NP合成接着马丁森和雷迪（2009）11的方法。 </li></ol> 3.治疗PBW与氧化铜纳米粒<ol><li>加50毫克的CuO-NP（1毫克/毫升），以50毫升锥形管中随后50毫升PBW的。密封管，并在250转反应30分钟的长凳上顶轨道摇床。 </li><li>离心样品管在250×g离心30分钟，然后用0.45微米的注射器过滤器过滤上清液。改变离心机速度和时间可以取决于纳米颗粒，以确保的CuO纳米粒成为紧凑的离心管中。 </li></ol> 4.元素分析<ol><li>制备未处理（对照）和CuO-NP处理PBW样品进行元素分析如下。 </li><li>酸化的CuO-NP-处理和未处理PBW等份（40毫升）与痕量金属级硝酸至pH 2.0。对于阳离子通过感应coupl分析酸化PBW等分编等离子体-质谱（ICP-MS），如雷迪和Roth（2012）13中所述。 </li><li>制备的CuO-NP-处理和未处理PBW的未酸化等份（20毫升）并分析未酸化等份阴离子通过如雷迪和Roth（2012）13中所述的离子色谱法（IC）。 注：等分被农业部分析服务的怀俄明部门，怀俄明州拉勒米的82070. IC和等离子体质谱过程的描述可以在雷迪和罗斯，（2012）13中找到分析。 </li></ol> 5.制备细胞培养基使用PBW <ol><li>使用两个控制（EMEM-1x和RO +媒体）和八个PBW测试的媒体解决方案（四种浓度每未处理PBW和CuO-NP处理的介质）的可行性研究。是溶液的概述如下： <ol><li>对于EMEM-1X控制，购买的Eagle氏最低必需培养基（EMEM-1×）与L-谷氨酰胺和碳酸氢钠已经添加。添加胎牛血清（FBS）并且按照制造商的说明，抗生素。 注：EMEM-1X被购买稀释到适当的浓度对细胞生长和含L-谷氨酰胺和碳酸氢钠。 EMEM-1X需要添加胎牛血清（FBS）和青霉素和链霉素（50IU / ml青霉素和50μg/ ml链霉素）的抗生素混合物的。 EMEM-1X被用作对照培养基，因为它是在制造商推荐的生长培养基在该研究中使用两种类型的细胞。浓缩EMEM-10倍，从设施或未经处理或的CuO-NP处理PBW稀释RO水，以产生测试溶液。浓缩EMEM-10倍时购买不包含所以这些被添加除了胎牛血清（FBS）和青霉素和链霉素的抗生素混合物L-谷氨酰胺或碳酸氢钠。 </li><li>对于RO控制解决方案中使用RO水从ISR收集设施。使用相同的协议PBW测试仅媒体替代100％RO笏呃从地方PBW的ISR设施。从实验室稀未处理和CuO-NP处理液使用的RO或超纯水。 </li><li>稀未处理PBW为四个测试浓度与细胞培养基成分混合之前。通过在下列组合混合未处理PBW用RO（从实验室）制备的4种不同浓度的未处理PBW溶液：100％（纯PBW +无RO水），75％（187.5毫升PBW +62.5毫升RO水的）， 50％（125毫升PBW + 125毫升RO水的）或25％（62.5毫升PBW +187.5毫升的RO水）。 </li><li>稀的CuO-NP处理PBW为四个测试浓度与细胞培养基成分混合之前。在下列组合通过混合PBW制备的4种不同浓度的的CuO-NP处理PBW溶液（预处理用1毫克/毫升的CuO-NP，30分钟）用RO（从实验室）：100％（纯CuO- NP处理PBW +无RO水），75％（187.5毫升的PBW +62.5毫升RO水的CuO-NP处理的），50％（125毫升的CuO-NP处理PBW + 125毫升RO水）或25％（62.5毫升的CuO-NP处理PBW +187.5毫升的RO水）。 </li></ol></li><li>加入25毫升浓EMEM-10倍的190毫升100％的RO及100％，75％，50％或准备250毫升反渗透+媒体，未处理PBW +媒体和CuO-NP处理PBW +媒浓度在步骤6.1.3和6.1.4中创建的预制未处理的或氧化铜-NP处理PBW浓度的25％。 </li><li>调节每个溶液至7.4，用NaOH或HCl的pH。 </li><li>补充未处理和CuO-NP处理PBW的各个浓度以及反渗透+媒体具有以下标准组件：25毫升（10％）胎牛血清（FBS），2.5毫升L-谷氨酰胺，0.55克碳酸氢钠和1.25ml笔/链霉素（50IU / ml青霉素和50μg/ ml链霉素）。 </li><li>调整未处理PBW +介质的每个浓度的渗透压，加入RO水，并用渗透压计测量的CuO-NP处理PBW +媒体以及RO +媒体290-310毫渗透摩尔浓度/千克。 </li><li>使用过滤器每个解决方案0.22微米真空过滤器单元，并储存在4℃。 注意：由于在RO水用于调节渗透压的量的轻微变化，变化在5％范围内的最终介质浓度，与未处理PBW +媒浓度在56％，44％，29％和16.5％，和CuO-NP-处理PBW +媒浓度为53％，45％，30％及17％。 </li></ol> 6.细胞活力注：由于肾脏和肝脏的重金属毒性的靶器官，采用培养的人类胚胎肾（HEK293）细胞（HEK）和人肝癌（肝癌）细胞（HEP）的测试方法24-26。 <ol><li>电镀在根据制造商的说明，将实验中使用的96孔板前2-3天制备HEK和HEP细胞的培养物。 </li><li>使用3- [4，5-二甲基吡啶-2-基] -2，5-二苯基溴化物（MTT）测定法测量细胞活力。 注：MTT法协议是从眉修改rloo 等人。（2011）27。 <ol><li>以粉末形式获得MTT。添加磷酸盐缓冲液（PBS），以弥补股票浓度为50毫克/毫升。搅拌该溶液2小时，然后过滤用0.45μm针筒式滤器和分装成1.5毫升冷冻安全管。避光并储存管在4℃下。 </li></ol></li><li>从用胰蛋白酶，离心其培养皿除去HEK和Hep细胞在1000×g离心5分钟，并滗出胰蛋白酶。加入5毫升的PBS和混合细胞以获得单细胞溶液。然后，应用将20μl单细胞溶液到血球以获得每毫升溶液细胞计数。再次离心细胞在1,000×g离心5分钟，倒出PBS用于冲洗细胞。添加EMEM-1倍的适量调节细胞浓度至500个细胞/ 100微升（100μl/孔）。 </li><li>填充板的周边的孔用200μlPBS中，以控制蒸发。 </li><li>种子细胞s的500个细胞/孔的密度，加入100微升至每个孔，除了外围孔（其不与细胞铺板）。 注：播种密度和HEK Hep细胞是基于实验的增长曲线，使经济增长的高峰期发生天左右4-5。准备生长曲线对所有细胞系来估计接种密度。 </li><li>孵育细胞24小时，在37℃下使它们进行细胞密度的基线MTT读数之前恢复（形式​​紧粘连至板）。 </li><li>通过从第一塔提取播种介质（不包括周边）并加入100微升的MTT（5mg / ml的在介质）向孔1小时进行细胞密度的基线MTT读数。 </li><li>一小时后，取出MTT法和加入100微升二甲基亚砜（DMSO）的溶解由活细胞（20分钟）产生的MTT-甲。 </li><li>读出的第一列的光密度（OD）在570纳米的吸收波长，得到基行读取。 <ol><li>使用基准读数，以确保所有平板接种正确和细胞板之间持续增长。从塔中取出DMSO培养未来24小时前进行测试。 注意：如果二甲基亚砜留在板过夜它拉水分从相邻列，导致在介质体积的减少。 </li></ol></li><li>温暖的测试解决方案（ 即 EMEM-1X，RO，未经处理的PBW和CuO-NP处理PBW媒体解决方案），以37℃的水浴。 </li><li>从板（不包括周边或将其用于基线读数的第一列），并用100微升EMEM-1倍，反渗透+媒体，未处理PBW +媒浓度或的CuO-NP的取代的其余部分移除播种媒体 - 处理PBW +媒体浓度（每盘一个解决方案）。孵育细胞在他们的测试浓度或控制解决方案，共七天（天2-8）。 注：10共板块：1 EMEM-1X，1 RO +媒体，各1个未处理PBW +媒体浓度（56％，44％，29％和16.5％），每个的CuO-NP处理PBW +媒体集中的一个板块（53％，45％ ，每细胞系实验的30％和17％）。 </li><li>每天以下基准MTT阅读，从各自板的下一列取下控制和测试解决方案（下6.11的说明中列出）（ 如第2天测试和控制媒体从第3行中删除，BG井;第3天：行4，井BG 等 ），并重复MTT协议为在上述步骤6.7-6.9所述。 </li><li>每天重复协议七天。平均外径结果的每一行（6孔），并报告与时间，以产生一个七天的生长曲线。 </li><li>为了评估铜螯合的对细胞活力的影响的CuO-NP处理PBW +媒体按照相同的步骤如上述，除了加入的解决方案，以它们各自的板之前添加的D-青霉胺100μM的控制和测试方案。执行数据肛ysis用科学绘图软件。 </li></ol> 7.地球化学模拟<ol><li>从以下网站下载的Visual MINTEQ 3.0 / 3.1一个免费<a href="http://www2.lwr.kth.se/English/Oursoftware/vminteq/">http://www2.lwr.kth.se/English/Oursoftware/vminteq/</a> 。 注：Visual MINTEQ是一个免费的化学平衡模型的金属形态，溶解平衡，吸附等天然水域的计算。此外，它是用来预测离子物种，离子活动，离子络合物和饱和指数是相对于元件的前，后处理（质谱结果）的浓度，以检查元件除去28的可能机制。 </li><li>打开程序和输入从第4步，包括pH值，碱度和不同的元素的浓度，进入程序的质谱数据。 注：由于地下水的过程中就地 urani氧化嗯提取工艺，用砷，钒和铀的氧化物质为输入。 </li></ol> 8.抑制浓度50（IC 50） <ol><li>由第一平均上的三个独立的运行5天的存活率（OD平均值）计算的IC 50为未处理和CuO-NP处理PBW +介质的浓度。 </li><li>减去第五天从生存能力EMEM-1X的第五天生存能力平均未处理和CuO-NP处理PBW +媒体浓度的平均值来计算可行性的差异。然后除以在EMEM第5天的存活率差异由平均存活率，再乘以100得到的抑制百分比。 </li><li>减去100（EMEM-1X生存能力）抑制百分比，以获取每个未处理和CuO-NP处理PBW +媒体集中的百分比生存能力。 </li><li>输入到通过设置EMEM-1x在一个的浓度和为100％的存活率科学绘图软件;全部改造成浓度日志规模（X =日志（X）），并进行非线性最小二乘回归拟合分析。 </li></ol> 9.数据分析<ol><li>在未经处理的和CuO-NP处理PBW元素的浓度与双尾成对，学生T检验进行比较。 </li><li>通过使用收集的七天增长曲线的数据计算的曲线（AUC）下面积，并分析与重复测量方差分析（ANOVA）的方差，然后所有组之间Tukey的事后比较（n = 3时）。 </li><li>通过使用数据从生长曲线的五天为未处理和CuO-NP处理PBW +媒体溶液（如上所述）计算IC 50值。对&lt;0.05被认为是显著P值。 注：统计分析的目的，一半的检出限质谱值被分配到离子浓度水平低于该限制29。 </li></ol>

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Molecular, Clinical and Environmental Toxicology. 101, 133-164 (2012).</li><li>Meerloo, J., Kaspers, G., Cloos, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+Sensitivity+Assays:+The+MTT+Assay.">Cell Sensitivity Assays: The MTT Assay.</a> Cancer Cell Culture. 731, 237-245 (2011).</li><li>Gustafsson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Visual MINTEQ. , (2010).</li><li>Hallab, N., Caicedo, M., McAllister, K., Skipor, A., Amstutz, H., Jacobs, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asymptomatic+prospective+and+retrospective+cohorts+with+metal-on-metal+hip+arthroplasty+indicate+acquired+lymphocyte+reactivity+varies+with+metal+ion+levels+on+a+group+basis.">Asymptomatic prospective and retrospective cohorts with metal-on-metal hip arthroplasty indicate acquired lymphocyte reactivity varies with metal ion levels on a group basis.</a> Journal of Orthopaedic Research. 31 (2), 173-182 (2013).</li><li>Goswami, A., Raul, P., Purkait, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arsenic+adsorption+using+copper+(II)+oxide+nanoparticles.">Arsenic adsorption using copper (II) oxide nanoparticles.</a> Chemical Engineering Research and Design. 90 (9), 1387-1396 (2011).</li><li>Pillewan, P., Mukherjee, S., Roychowdhury, T., Das, S., Bansiwal, A., Rayalu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Removal+of+As(III)+and+As(V)+from+water+by+copper+oxide+incorporated+mesoporous+alumina.">Removal of As(III) and As(V) from water by copper oxide incorporated mesoporous alumina.</a> Journal of Hazardous Materials. 186 (1), 367-375 (2011).</li><li>Kroll, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytotoxicity+screening+of+23+engineered+nanomaterials+using+a+test+matrix+of+ten+cell+lines+and+three+different+assays.">Cytotoxicity screening of 23 engineered nanomaterials using a test matrix of ten cell lines and three different assays.</a> Particle and fibre toxicology. 8 (9), 1-19 (2011).</li><li>Fahmy, B., Cormier, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Copper+oxide+nanoparticles+induce+oxidative+stress+and+cytotoxicity+in+airway+epithelial+cells.">Copper oxide nanoparticles induce oxidative stress and cytotoxicity in airway epithelial cells.</a> Toxicology in vitro: an international journal published in association with BIBRA. 23 (7), 1365-1371 (2009).</li><li>Radike, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distribution+and+accumulation+of+a+mixture+of+arsenic,+cadmium,+chromium,+nickel+and+vanadium+in+mouse+small+intestin,+kidney,+pancreas,+and+femur+following+oral+administration+in+water+or+feed.">Distribution and accumulation of a mixture of arsenic, cadmium, chromium, nickel and vanadium in mouse small intestin, kidney, pancreas, and femur following oral administration in water or feed.</a> Journal of Toxicology and Environmental Health, Part A. 65 (23), 2029-2052 (2002).</li><li>Barbier, O., Jacquillet, G., Tauc, M., Cougnon, M., Poujeol, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+heavy+metals+on,+and+handling+by,+the+kidney.">Effect of heavy metals on, and handling by, the kidney.</a> Nephron. Physiology. 99 (4), 105-110 (2005).</li><li>Zheng, X., Watts, G., Vaught, S., Gandolfi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-level+arsenite+induced+gene+expression+in+HEK293+cells.">Low-level arsenite induced gene expression in HEK293 cells.</a> Toxicology. 187 (1), 39-48 (2003).</li><li>Li, Z., Piao, F., Liu, S., Wang, Y., Qu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subchronic+exposure+to+arsenic+trioxide-induced+oxidative+DNA+damage+in+kidney+tissue+of+mice.">Subchronic exposure to arsenic trioxide-induced oxidative DNA damage in kidney tissue of mice.</a> Experimental and Toxicologic Pathology. 62 (5), 543-547 (2010).</li><li>Farombi, E., Akintunde, J., Nzute, N., Adedara, I., Arojojoye, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Municipal+landfill+leachate+induces+hepatotoxicity+and+oxidative+stress+in+rats.">Municipal landfill leachate induces hepatotoxicity and oxidative stress in rats.</a> Toxicology and Industrial Health. 28 (6), 532-541 (2011).</li><li>Das, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arsenic+exposure+through+drinking+water+increases+the+risk+of+liver+and+cardiovascular+diseases+in+the+population+of+West+Bengal.">Arsenic exposure through drinking water increases the risk of liver and cardiovascular diseases in the population of West Bengal.</a> India. BMC public health. 12 (1), 639-648 (2012).</li><li>Valko, M., Morris, H., Cronin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metals,+toxicity+and+oxidative+stress.">Metals, toxicity and oxidative stress.</a> Current Medicinal Chemistry. 12 (10), 1161-1208 (2005).</li><li>Horie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+Adsorption+of+Ultrafine+Metal+Oxide+and+Its+Influence+on+Cytotoxicity+toward+Cultured+Cells.">Protein Adsorption of Ultrafine Metal Oxide and Its Influence on Cytotoxicity toward Cultured Cells.</a> Chemical Research in Toxicology. 22 (3), 543-553 (2009).</li></ol>

The authors have nothing to disclose.

以前的研究报告说，氧化铜，纳米粒子去除砷地下水11,13,30,31。这项研究支持，这些以往的调查结果也报告说，氧化铜纳米粒删除PBW更多的污染物。本研究还证实了以前的报告的CuO纳米粒是在除砷有效，尽管其他污染物和可能的竞争离子11的存在。形态建模预测，钒物种PBW的97％的负电荷，从而允许吸附在CuO纳米粒和批量处理除去钒的92％。 这是第一次研究，?...

大约是美国电力供应的20％是由核能，部分基于国家鼓励提高能源独立性提供，预计美国的核能力，以增加1。核能的全球增长预计也将继续，与许多美国以外的2出现的增长。截至2013年，美国铀的83％是进口的，但储量952544吨，在美国3,4存在。在2013年有7个新设施的应用和怀俄明州，新墨西哥州，内布拉斯加州和5之间14重启/扩展应用。在美国，铀为主通过原位恢复（ISR）处理6萃取。 ISR导致更少的土地破坏和避免造成尾矿堆，可以释放环境污染物7。 ISR使用水基氧化解决方案，以从​​地下矿石体，在这之后的铀从浸出液通过提取浸出铀离子交换过程8。维持负面水平衡在矿体，浸出液的一部分，被称为生产流血水（PBW）中，放血关闭。所述PBW的一部分被利用反渗透（RO）和净化重新引入开采过程中，但PBW也可以有利于工业或农业用途，如果有毒的污染物可以减少到由状态管理机构对于表面确定的可接受水平和地下水9。目前，大多数ISR铀的设施使用RO从PBW去除污染物。然而，反渗透处理是能源密集型，并产生有毒废物盐水，这需要监管的处置。 许多水净化方法都存在，包括吸附剂，膜和离子交换。在这些中，吸附是最常用的，并且最近的发展纳米颗粒合成具有增强的吸附剂基水净化处理10的功能。铜OXI德纳米颗粒（氧化铜纳米粒）以前没有被广泛研究对铀的ISR PBW，但在最近的污染物去除从地下水的研究，氧化铜纳米粒被发现有独特的性能，包括无需预处理或后处理的水处理工序（ 例如 ，调节pH值或氧化还原电位），并在不同的水成分表现良好（ 例如 ，在不同的pH值，盐的浓度，或竞争离子）11。另外，氧化铜纳米粒容易通过用氢氧化钠（NaOH），之后，将再生的CuO纳米粒可重复使用浸出再生。从天然水域的CuO-NP微量金属过滤功能细节已先前公布11-14。 虽然对水处理是有用的，金属氧化物纳米颗粒可能是有毒的活生物体，但毒性的程度取决于，部分地基于纳米粒子的特性和成分10,15,16。因此，研究SIMULT是很重要现场应用之前aneous污染物的去除和纳米粒子毒性。目前的研究确定的CuO纳米粒的能力以去除PBW优先污染物（包括砷，硒，钒和铀），和评估的CuO-NP处理对PBW细胞毒性的影响。 PBW从有源ISR铀设施收集并用于确定在优先的污染物去除的CuO-NP处理的功效。之前和之后的CuO-NP处理PBW细胞毒性也进行了评估。 PBW是一个复杂的地质（工业/环境）的混合物和环境都和健康科学（NIEHS）国家研究所和机构的有毒物质及疾病登记（ASTDR）是把重点放在学习环境相关的混合物，包括混合物的毒性因为它们在自然界或工业设置，以及在体外测试促进存在优先的化学品进行进一步的体内测试17-19。是慢性，低剂量暴露的混合物具有挑战性的研究，因为长期接触低剂量的混合物，不会产生明显的影响，至少在大多数实验室研究的很短的时间框架。同样，大多数在化学混合物的体外研究细胞暴露于两种或更多种金属20,21限定的实验室制造的混合物。这些研究提供基线信息，但简化的混合物不复制，可能会出现一个天然的，环境样品，其中所述全方位混合物组分都存在于该复合拮抗和协同相互作用。 本研究的目标是检查供PBW备用污染物去除工艺和评估（的CuO-NP）的治疗上PBW细胞毒性使用培养的人类细胞的效果。该结果可能通过更有效的或环保的方法来去除污染物的发展中受益的铀业。这项研究提供第一个证据是PBW减少污染物优先由氧化铜纳米粒在哺乳动物细胞中22毒性降低。

<table border="0" cellpadding="0" cellspacing="0" width="892">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:243px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:315px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">CuCl2</td>
			<td style="width:243px;">Sigma</td>
			<td align="right" style="width:119px;">203149</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Borosilicate glass balls</td>
			<td style="width:243px;">VWR</td>
			<td>26396-639</td>
			<td>6 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Nitric Acid</td>
			<td style="width:243px;">Fisher</td>
			<td>A509-P500</td>
			<td>Trace metal grade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.45 mm syringe filter</td>
			<td style="width:243px;">Fisher</td>
			<td style="width:119px;">SLHA 033S S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">10X EMEM</td>
			<td style="width:243px;">Fisher</td>
			<td style="width:119px;">BW12-684F</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fetal Bovine Serum</td>
			<td style="width:243px;">ATCC</td>
			<td>30-2020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">L-glutamine</td>
			<td style="width:243px;">Fisher</td>
			<td style="width:119px;">BP379-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">NaHCO3</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:119px;">S5761</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Penicillin/Streptomycin</td>
			<td style="width:243px;">ATCC</td>
			<td style="width:119px;">30-2300</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.22 mm vacuum filter unit</td>
			<td style="width:243px;">Fisher</td>
			<td style="width:119px;">09-740-28C</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HEK293</td>
			<td style="width:243px;">ATCC</td>
			<td style="width:119px;">CRL-1573</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HEPG2</td>
			<td style="width:243px;">ATCC</td>
			<td style="width:119px;">HB-8065</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Trypsin</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:119px;">SV3003101</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">MTT</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:119px;">M2128</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">D-penicillamine</td>
			<td style="width:243px;">Fisher</td>
			<td style="width:119px;">ICN15180680</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">96-well plates</td>
			<td style="width:243px;">Fisher</td>
			<td style="width:119px;">07-200-92</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">DMSO</td>
			<td style="width:243px;">Fisher</td>
			<td style="width:119px;">D12814</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Spectra Max 190</td>
			<td style="width:243px;">Molecular Devices</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Visual MINTEQ version 3.0</td>
			<td style="width:243px;">KTH Royal Institute of Technology</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">ICP-MS&#160;</td>
			<td style="width:243px;">Agilent</td>
			<td style="width:119px;"></td>
			<td>Details of instruments, models and detection limits were published in Reddy et al., 2013.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">IC DIONEX DX 500</td>
			<td style="width:243px;">Dionex</td>
			<td style="width:119px;"></td>
			<td>Details of instruments, models and detection limits were published in Reddy et al., 2013.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">VWR Incubator</td>
			<td style="width:243px;">VWR</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

removal of trace elements by cupric oxide nanoparticles from uranium in situ recovery bleed water and its effect on cell viability

在原位恢复（ISR）是提取铀在美国的主要方法。期间的ISR，铀是从矿体浸出，并通过离子交换萃取。所得的生产泌水（PBW）含有污染物，如砷等重金属。 PBW从激活ISR铀设施处理样品与氧化铜纳米粒子（氧化铜纳米粒）。的CuO-NP处理PBW降低优先污染物，包括砷，硒，铀，和钒。未处理和CuO-NP处理通过MTT（3-（4,5-二甲基吡啶-2-基）-2,5-二苯基溴化）测定PBW用作细胞生长介质和变化生存能力的液体成分分析在人胚肾（HEK 293）和人肝细胞癌（Hep G2细胞）细胞。的CuO-NP处理用改进的HEK和HEP细胞活力有关。这种方法的限制包括稀释PBW由生长培养基成分和渗透压克分子期间先进而精湛调整以及必要pH调节。这种方法是在其更广泛的范围，由于稀释作用以及变化PBW的pH是传统微酸但是限制;这种方法可以有更广泛的用途评估的CuO-NP处理的较为中性的水域。

PBW成分浓度和pH值在未处理和CuO-NP处理PBW报道于表1中 。马丁森和雷迪（2009）报告说，估计在9.4±0.4的零电荷的CuO-NP的点。鉴于PBW的pH为7.2-7.4，在这些条件下，水捐赠质子的CuO纳米粒，使纳米颗粒表面带正电荷，允许带负电荷的物质的吸附。从PBW的CuO-NP处理去除优先污染物，包括砷，硒，铀和钒（ 表1）。平均砷浓度降低了87％[从0.0175到0.002毫克/升（双尾配对t检验，p</...

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Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Production bleed water (PBW) was treated with cupric oxide nanoparticles (CuO-NPs) and cellular toxicity was assessed in cultured human cells. The goal of this protocol was to integrate the native environmental sample into a cell culture format assessing the changes in toxicity due to CuO-NP treatment.

去除微量元素的氧化铜纳米粉体制备铀<em&gt;原位</em&gt;恢复泌水及其对细胞活力的影响

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted ...

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Research

JoVE Journal

Environment

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

9.7K 次观看.  University of New Mexico. Production bleed water (PBW) was treated with cupric oxide nanoparticles (CuO-NPs) and cellular toxicity was assessed in cultured human cells. The goal of this protocol was to integrate the native environmental sample into a cell culture format assessing the changes in toxicity due to CuO-NP treatment.

视频: Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Laboratory Estimation of Net Trophic Transfer Efficiencies of PCB Congeners to Lake Trout (Salvelinus namaycush) from Its Prey

A technique for laboratory estimation of net trophic transfer efficiency (&#947;) of polychlorinated biphenyl (PCB) congeners to piscivorous fish from their prey is described herein. During a 135-day laboratory experiment, we fed bloater (Coregonus hoyi) that had been caught in Lake Michigan to lake trout (Salvelinus namaycush) kept in eight laboratory tanks. Bloater is a natural prey for lake trout. In four of the tanks, a relatively high flow rate was used to ensure relatively high activity by the lake trout, whereas a low flow rate was used in the other four tanks, allowing for low lake trout activity. On a tank-by-tank basis, the amount of food eaten by the lake trout on each day of the experiment was recorded. Each lake trout was weighed at the start and end of the experiment. Four to nine lake trout from each of the eight tanks were sacrificed at the start of the experiment, and all 10 lake trout remaining in each of the tanks were euthanized at the end of the experiment. We determined concentrations of 75 PCB congeners in the lake trout at the start of the experiment, in the lake trout at the end of the experiment, and in bloaters fed to the lake trout during the experiment. Based on these measurements, &#947; was calculated for each of 75 PCB congeners in each of the eight tanks. Mean &#947; was calculated for each of the 75 PCB congeners for both active and inactive lake trout. Because the experiment was replicated in eight tanks, the standard error about mean &#947; could be estimated. Results from this type of experiment are useful in risk assessment models to predict future risk to humans and wildlife eating contaminated fish under various scenarios of environmental contamination.

Laboratory Estimation of Net Trophic Transfer Efficiencies of PCB Congeners to Lake Trout Salvelinus namaycush from Its Prey

A technique for laboratory estimation of net trophic transfer efficiency of polychlorinated biphenyl (PCB) congeners to piscivorous fish from their prey is presented. To maximize applicability of the laboratory results to the field, the piscivorous fish should be fed prey fish that are typically eaten in the field.

多氯联苯同系物净营养转移效率湖鳟鱼实验室估计（<em&gt;红点鲑namaycush</em&gt;），从它的猎物

A technique for laboratory estimation of net trophic transfer efficiency (&#947;) of polychlorinated biphenyl (PCB) congeners to piscivorous fish from ...

科研

环境科学

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) ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. The standardized sampling component of the method concentrates viruses that may be present in water by passage of a minimum specified volume of water through an electropositive cartridge filter. The minimum specified volumes for surface and finished/ground water are 300 L and 1,500 L, respectively. A major method limitation is the tendency for the filters to clog before meeting the sample volume requirement. Studies using two different, but equivalent, cartridge filter options showed that filter clogging was a problem with 10% of the samples with one of the filter types compared to 6% with the other filter type. Clogging tends to increase with turbidity, but cannot be predicted based on turbidity measurements only. From a cost standpoint one of the filter options is preferable over the other, but the water quality and experience with the water system to be sampled should be taken into consideration in making filter selections.

EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.

EPA方法1615测量肠道病毒和诺瓦克病毒发生水由文化和RT-qPCR的了。病毒样本的收集I.

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a standardized method for use in multiple analytical laboratories during monitoring period 3 of the Unregulated Contaminant Monitoring Rule. Herein we present the protocol for extraction of viral ribonucleic acid (RNA) from water sample concentrates and for quantitatively measuring enterovirus and norovirus concentrations using reverse transcription-quantitative PCR (RT-qPCR). Virus concentrations for the molecular assay are calculated in terms of genomic copies of viral RNA per liter based upon a standard curve. The method uses a number of quality controls to increase data quality and to reduce interlaboratory and intralaboratory variation. The method has been evaluated by examining virus recovery from ground and reagent grade waters seeded with poliovirus type 3 and murine norovirus as a surrogate for human noroviruses. Mean poliovirus recoveries were 20% in groundwaters and 44% in reagent grade water. Mean murine norovirus recoveries with the RT-qPCR assay were 30% in groundwaters and 4% in reagent grade water.

Here we present a procedure to quantify enterovirus and norovirus in environmental and drinking waters using reverse transcription-quantitative PCR. Mean virus recovery from groundwater with this standardized procedure from EPA Method 1615 was 20% for poliovirus and 30% for murine norovirus.

EPA方法1615测量肠道病毒和诺瓦克病毒发生水由文化和RT-qPCR的了。第三部分。病毒检测用RT-qPCR的

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a ...

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 ...

Determination of Inorganic Arsenic in a Wide Range of Food Matrices using Hydride Generation - Atomic Absorption Spectrometry.

The European Food Safety Authority (EFSA) underlined in its Scientific Opinion on Arsenic in Food that in order to support a sound exposure assessment to inorganic arsenic through diet, information about distribution of arsenic species in various food types must be generated. A method, previously validated in a collaborative trial, has been applied to determine inorganic arsenic in a wide variety of food matrices, covering grains, mushrooms and food of marine origin (31 samples in total). The method is based on detection by flow injection-hydride generation-atomic absorption spectrometry of the iAs selectively extracted into chloroform after digestion of the proteins with concentrated HCl. The method is characterized by a limit of quantification of 10 &#181;g/kg dry weight, which allowed quantification of inorganic arsenic in a large amount of food matrices. Information is provided about performance scores given to results obtained with this method and which were reported by different laboratories in several proficiency tests. The percentage of satisfactory results obtained with the discussed method is higher than that of the results obtained with other analytical approaches.

The usefulness of an analytical method to determine inorganic arsenic in a wide range of food matrices is demonstrated. The method consists of selective extraction of inorganic arsenic into chloroform with a final determination by hydride generation-atomic absorption spectrometry.

用氢化物发生-原子吸收光谱法测定多种食品基质中的无机砷。

The European Food Safety Authority (EFSA) underlined in its Scientific Opinion on Arsenic in Food that in order to support a sound exposure assessment to ...

Experimental Column Setup for Studying Anaerobic Biogeochemical Interactions Between Iron (Oxy)Hydroxides, Trace Elements, and Bacteria

Fate and speciation of trace elements (TEs), such as arsenic (As) and mercury (Hg), in aquifers are closely related to physio-chemical conditions, such as redox potential (Eh) and pH, but also to microbial activities that can play a direct or indirect role on speciation and/or mobility. Indeed, some bacteria can directly oxidize As(III) to As(V) or reduce As(V) to As(III). Likewise, bacteria are strongly involved in Hg cycling, either through its methylation, forming the neurotoxin monomethyl mercury, or through its reduction to elemental Hg&#176;. The fates of both As and Hg are also strongly linked to soil or aquifer composition; indeed, As and Hg can bind to organic compounds or (oxy)hydroxides, which will influence their mobility. In turn, bacterial activities such as iron (oxy)hydroxide reduction or organic matter mineralization can indirectly influence As and Hg sequestration. The presence of sulfate/sulfide can also strongly impact these particular elements through the formation of complexes such as thio-arsenates with As or metacinnabar with Hg.
Consequently, many important questions have been raised on the fate and speciation of As and Hg in the environment and how to limit their toxicity. However, due to their reactivity towards aquifer components, it is difficult to clearly dissociate the biogeochemical processes that occur and their different impacts on the fate of these TE.
To do so, we developed an original, experimental, column setup that mimics an aquifer with As- or Hg-iron-oxide rich areas versus iron depleted areas, enabling a better understanding of TE biogeochemistry in anoxic conditions. The following protocol gives step by step instructions for the column set-up either for As or Hg, as well as an example with As under iron and sulfate reducing conditions.

Experimental Column Setup for Studying Anaerobic Biogeochemical Interactions Between Iron OxyHydroxides, Trace Elements, and Bacteria

Fate and speciation of arsenic and mercury in aquifers are closely related to physio-chemical conditions and microbial activity. Here, we present an original experimental column setup that mimics an aquifer and enables a better understanding of trace element biogeochemistry in anoxic conditions. Two examples are presented, combining geochemical and microbiological approaches.

用于研究铁 (氧) 氢氧化物、微量元素和细菌之间厌氧生物地球化学相互作用的实验柱装置

Fate and speciation of trace elements (TEs), such as arsenic (As) and mercury (Hg), in aquifers are closely related to physio-chemical conditions, such as ...

Assessment of Methane and Nitrous Oxide Fluxes from Paddy Field by Means of Static Closed Chambers Maintaining Plants Within Headspace

This protocol describes the measurement of greenhouse gas (GHG) emissions from paddy soils using the static closed chamber technique. This method is based on the diffusion theory. A known volume of air overlaying a defined soil area is enclosed within a parallelepiped cover (named &#34;chamber&#34;), for a defined period of time. During this enclosure period, gases (methane (CH4) and nitrous oxide (N2O)) move from soil pore air near their microbial source (i.e., methanogens, nitrifiers, denitrifiers) to the chamber headspace, following a natural concentration gradient. Fluxes are then estimated from chamber headspace concentration variations sampled at regular intervals throughout the enclosure and then analyzed with gas chromatography. Among the techniques available for GHG measurement, the static closed chamber method is suitable for plot experiments, as it does not require large homogenously treated soil areas. Furthermore, it is manageable with limited resources and can identify relationships among ecosystem properties, processes, and fluxes, especially when combined with GHG driving force measurements. Nevertheless, with respect to the micrometeorological method, it causes a minimal but still unavoidable soil disturbance, and allows a minor temporal resolution. Several phases are key to the method implementation: i) chamber design and deployment, ii) sample handling and analyses, and iii) flux estimation. Technique implementation success in paddy fields demands adjustments for field flooding during much of the cropping cycle, and for rice plant maintenance within the chamber headspace during measurements. Therefore, the additional elements to be considered with respect to the usual application of non-flooded agricultural soils consist of devices for: i) avoiding any unintended water disturbance that could overestimate fluxes, and ii) including rice plants within the chamber headspace to fully consider gases emitted through aerenchyma transportation.

The overall goal of this protocol is to measure greenhouse gas emissions from paddy fields using the static closed chamber technique. The measurement system needs specific adjustments due to the presence of both a permanent water layer in the field and of the plants within the chamber headspace.

用静止封闭室维持顶空装置对稻田甲烷和一氧化二氮通量的评价

This protocol describes the measurement of greenhouse gas (GHG) emissions from paddy soils using the static closed chamber technique. This method is based ...

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol for performing in situ juvenile exposure tests within oligotrophic river catchments over one-month and three-month periods. Two methods (in both modifications) are presented to evaluate the juvenile growth and survival rate. The methods and modifications differ in value for the locality bioindication and each has its benefits as well as limitations. The sandy cage method works with a large set of individuals, but only some of the individuals are measured and the results are evaluated in bulk. In the mesh cage method, the individuals are kept and measured separately, but a low individual number is evaluated. The open water exposure modification is relatively easy to apply; it shows the juvenile growth potential of sites and can also be effective for water toxicity testing. The within-bed exposure modification needs a high workload but is closer to the conditions of a natural juvenile environment and it is better for reporting the real suitability of localities. On the other hand, more replications are needed in this modification due to its high-hyporheic environment variability.

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

In situ bioindications enable determination of the suitability of an environment for endangered mussel species. We describe two methods based on the juvenile exposure of freshwater pearl mussels in cages to oligotrophic river habitats. Both methods are implemented in variants for open water and hyporheic water environments.

用原位曝光法测定淡水珍珠贻贝 Bioindication 的流环境适宜性

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol ...

Assessing the Particulate Matter Removal Abilities of Tree Leaves

Based on the conventional cleaning methods (water cleaning (WC) + brush cleaning (BC)), this study evaluated the influence of ultrasonic cleaning (UC) on collecting various sized particulate matter (PM) retained on leaf surfaces. We further characterized the retention efficiency of leaves to various sized PM, which will help to assess the abilities of urban trees to remove PM from ambient air quantitatively. 
Taking three broadleaf tree species (Ginkgo biloba, Sophora japonica, and Salix babylonica) and two needleleaf tree species (Pinus tabuliformis and Sabina chinensis) as the research objects, leaf samples were collected 4 days (short PM retention period) and 14 days (long PM retention period) after the latest rainfall. PM retained on the leaf surfaces was collected by means of WC, BC, and UC in sequence. Then, retention efficiencies of leaves (AEleaf) to three types of the various sized PM, including easily removable PM (ERP), difficult-to-remove PM (DRP), and totally removable PM (TRP), were calculated. Only around 23%-45% of the total PM retained on leaves could be cleaned off and collected by WC. When the leaves were cleaned through WC+BC, the underestimation of the PM retention capacity of different tree species was in the range of 29%-46% for various sized PM. Almost all PM retained on leaves could be removed if UC was supplemented to WC+BC. 
In conclusion, if the UC was complemented after the conventional cleaning methods, more PM on leaf surfaces could be eluted and collected. The procedure developed in this study can be used for assessing the PM removal abilities of different tree species.

The ultrasonic cleaning method was applied to elute the particulate matter (PM) retained on leaf surfaces after PM was eluted by the conventional cleaning methods (water cleaning only or water cleaning plus brush cleaning). The methodology can help to improve the estimation accuracy for PM retention capacity of leaves.

树叶颗粒物去除能力的评价

Based on the conventional cleaning methods (water cleaning (WC) + brush cleaning (BC)), this study evaluated the influence of ultrasonic cleaning (UC) on ...