Our study was conducted with the permission of United States Sugar Corporation. We want to thank R. Raid, S. Raid and T. Lang for their assistant. We also M. Stanback and three anonymous reviewers for suggestions that improved this manuscript.

1.群陷阱盒建设注意:对于群陷阱建设两个备用协议包括在步骤1.1和1.2。有几种其他方式捕集箱可以构造。以下两种方法都在这里提出，因为我们已经在实验中对它们进行测试，他们获得了成功。对于这两种协议，请使用1.3厘米至2厘米厚的非加压处理的胶合板。 <ol><li>群陷阱协议选项1建设:构建一个陷阱箱<ol><li>切割6块夹板30厘米×30厘米</li><li>用钉子或螺丝将盒子的四边在一起。 </li><li>通过安装钉子或螺丝底部件。 </li><li>连接顶部片用螺丝或钉。 <ol><li>不要用钉来固定顶部。顶端必须是可移动的，以得到蜂出并清洁箱。 </li></ol></li><li>钻在陷阱框从底边缘的2.5 cm一个面的中央2.5cm的孔。 </li><li>应用底漆，Sealant，或油漆向告警框以保护木材免受天气的外侧。 <ol><li>不适用任何油漆或密封剂的陷阱箱的内部。这可以防护剂或有毒的蜜蜂。 </li></ol></li></ol></li><li>标准深蜂巢体的使用:群陷阱盒协议选项2的构建<ol><li>从本地蜜蜂供应商店获得一个标准的10帧深蜂巢的身体。 </li><li> 41.3厘米宽剪切从胶合板顶部和底部各长测量50.8厘米。 </li><li>通过安装钉子或螺丝底部件。 </li><li>连接顶部片用螺丝或钉。 <ol><li>不要用钉来固定顶部。顶端必须是可移动的，以得到蜂出并清洁箱。 </li></ol></li><li>钻上的陷阱框从底边缘的2.5 cm一侧的中间2.5cm的孔。 </li><li>应用底漆，密封剂或涂料的陷阱框以保护木材免受天气的外侧。 <ol><li>不要使用任何涂料或密封胶的里面的陷阱箱。这可以防护剂或有毒的蜜蜂。 </li></ol></li></ol></li></ol> 2.信息素诱剂的制备<ol><li>比例为1:在一个2制备柠檬醛和香叶醇的混合物。 </li><li>将1ml该混合物成一个管理单元顶部1.5毫升聚丙烯微量离心管中。 </li><li>诱惑包裹铝箔以保护他们免受光。 注意:当它被放置在安装群捕集离心管的盖是用推针进行穿刺。 注意:鱼饵可以存储在室温暗处或更长存储在-10℃冰箱中。 </li></ol> 3.群陷阱盒安装注:对于安装的步骤3.1和3.2给出群陷阱箱两个备用协议。 <ol><li>安装在一个帖子:群陷阱盒协议选项1的安装<ol><li>将10cm的10 cm2的压力处理木材栅栏后进入离地面10米至15米从那里鸟巢箱的位置。 </li><li>建设1.3厘米厚的压力处理的木材吊板通过20厘米测量20cm的</li><li>对准吊板在阱中的一侧的中间，以10厘米以上的盒的顶部。固定板箱至少有两个钉子或螺丝钉在板的两侧。 </li><li>放置在后的陷阱框的高度（1.5米到3米）时的陷阱箱蜜蜂殖民即安全又容易去除。由挂板安装螺钉框后。放置一个螺钉下面吊板的顶部大约5厘米，代替螺钉。 <ol><li>因为它们是难以去除，不要用指甲。 </li></ol></li></ol></li><li>安装在树上:群陷阱盒协议选项2的安装<ol><li>钻在方块的各边的后上角5毫米的孔。 </li><li>螺纹穿过孔的线。 注意:如果使用此协议将是EASI器通过前固定框的顶部中的孔放置的导线。 </li><li>选择一棵树约为10米至15米的距离从那里鸟巢箱的位置。 <ol><li>不要放置陷阱箱子在同一树木禽箱，因为当有人在鸟类中的状态检查，蜜蜂殖民陷阱框可能被干扰导致蜜蜂成为防御性​​的。 </li></ol></li><li>固定电线周围的树1.5米到3米离开地面。 注意:为了保护树，形成层可以节省导线周围使用。 <ol><li>确保框是水平和安全的，以便它不会翻倒。盒子可以在树枝上的如果有增加稳定性来休息。 </li></ol></li></ol></li></ol> 4.增加信息素诱<ol><li>取从第2预先制备引诱和穿刺管的顶部用推针。 </li><li>放置陷阱箱内的诱惑附近后出师表入口底部allation，以确保它不会掉出来，而安装的框。 </li></ol> 5.用苄氯菊酯鸟巢箱治疗<ol><li>选择只标明用于家禽使用或笼中鸟苄氯菊酯产品。 </li><li>按照标签上的说明进行混合和应用 注意:请按照标签建议的任何个人防护指导原则。 </li><li>喷巢箱的内部完全，直到该液体逃跑用3.7 l手泵式喷雾器的框的侧面的点。 注:巢箱可之前或在安装后进行处理。 </li></ol> 6.去除蜜蜂占据的陷阱箱<ol><li>插头用小毛巾洞口。 注意:穿适当的防护服，以防止蜜蜂蜇伤。 <ol><li>如果群是被杀死，这样做一旦被发现。如果群被利用，让蜜蜂试图删除它们之前完全殖民陷阱箱（Usually约2周）。如果可能的话，在夜间占用取出盒子陷阱（当所有的蜜蜂都在里面）。陷阱入口可在夜间插上，然后箱子在白天第二天早上除去。 注:晚上蜜蜂周围工作时使用红色滤光。 </li></ol></li><li>拧下后盒或切断电线抱着箱子树。 </li><li>移动的方块到新的位置（至少4公里远），并从入口孔拉出毛巾。 </li></ol> 7.陷阱箱破坏蜂群注:在它是非法的，以保持非洲化蜜蜂领域，殖民地的破坏可能有必要。如果是这样，两个协议是这里介绍。 <ol><li> Solarizing殖民地<ol><li>插头用小毛巾洞口。 注意:穿适当的防护服，以防止蜜蜂蜇伤。 </li><li>拧下后盒或切断电线抱着箱子树。 </li><li> EncloSE和完全密封的盒子陷阱在一个塑料袋，并在充满阳光留下了好几天 注意:穿适当的防护服，以防止蜜蜂蜇伤。 </li><li>通过去除死蜜蜂和梳子清洁开箱。 </li></ol></li><li>一个喷雾杀虫剂使用<ol><li>使用标示使用喷雾杀虫剂杀死蜜蜂陷阱箱内喷雾。 注意:遵循杀虫剂标签上的说明，并使用适当的个人防护装备，以防止蜜蜂蜇伤。 </li><li>堵塞入口和离开箱中放置24小时。 </li><li>通过去除死蜜蜂和梳子清洁开箱。 注意:要重用陷阱框内部清理用肥皂水溶液以去除残留的杀虫剂。用清水冲洗并允许重新安装之前完全干燥。 </li></ol></li><li>杀虫剂泡沫的用途<ol><li>填充占用阱的整个腔与基于洗涤剂泡沫。 （注意:请按照非正规部门服务标签上的说明ticide和使用适当的个人防护装备和佩戴合适的防护服，以防止蜜蜂蜇伤）。 注意:泡沫可与商业虫害控制泡沫发生器或压缩空气灭火器被应用。 注意:对于使用灭火器:将3.8升的水倒入9.5大号灭火器，加入190毫升发泡剂（或餐具洗涤剂）的，密封的灭火器，并加压至100-125英镑/平方米。英寸（psi）。只是使用，大力搅拌，然后应用泡沫，直到陷阱四溢之前。 注:泡沫通过进入气孔和阻塞气管杀死60秒内的蜜蜂。 </li><li>堵塞入口和离开箱中放置24小时。 </li><li>通过去除死蜜蜂和梳子清洁开箱。 注意:要重用陷阱框内部清理用肥皂水溶液以去除残留的杀虫剂。用清水冲洗并允许重新安装之前完全干燥。&lt;/ LI&gt; </ol></li></ol>

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The authors have nothing to disclose.

作为一个非常成功的入侵害虫17，AHB构成了整个范围腔筑巢的鸟类的主要竞争对手。这里介绍的协议，旨在通过AHB减少燕窝盒定植率。这种设计的主要组成部分包括:1）使用防护剂和鸟类安全的杀虫剂，氯菊酯的; 2）备用筑巢蜜蜂的规定; 3）使用内部的陷阱箱引诱信息素。选择一个新的巢址18时，蜜蜂通常把"正确"的决定。制作陷阱盒比鸟巢箱的吸引力将增加选择了前者存在的可能性。具体地说，陷阱箱应具有位于比天花板23更靠近地板较小的入口孔。最后，较高的箱子往往被蜜蜂是首选。然而，由于AHB远约巢址比EHB 2少挑剔，信息素是attr的最重要的组成部分活性24。 这个协议可以适于配合各种情况。对于蜜蜂的密度很高的地区，可以安装每个巢箱两个陷阱。对于悬崖筑巢的鸟类，陷阱可吊装到位平行于窝腔或上方和下方。陷阱箱可以放置在滑轮系统更容易去除那些难以到达的区域。非洲化蜜蜂有蜂拥赛季和一个赛季的潜逃与位置变化而变化。蜂拥通常与花的高峰花期一致。当潜逃蜜蜂的整个殖民地放弃巢址，通常是由于巢址的缺乏或发生变质。据预测，当这些季节是很重要的。因为蜂拥和潜逃因人而异的定时和频率，而且往往不文献报道，人们应该联系本地养蜂或农业机构的信息。这将允许一氯菊酯更有针对性的应用ND /或诱惑。此外，管理计划是该协议的成功是至关重要的。占据陷阱箱需要定期监测（至少每3周），搬迁或销毁。该协议是有效的可用陷阱箱蜜蜂拓殖的维护是非常重要的。总的目标是不降低蜜蜂的人口在一个地区，而是给他们的替代筑巢地点。 安全始终是一个优先事项。例如，尽管蜜蜂一般都比较喜欢高巢址，我们倾向于把陷阱箱低到地上，以方便他们安全移除。陷阱箱应放置在不会放置一个生物学家在检查危险燕窝中的状态位置。个人防护装备（蜂西装或夹克衫和蜜蜂手套）应该被任何人删除与蜜蜂里面陷阱箱佩戴。 我们在这项研究中，因为我使用了氯菊酯微胶囊喷雾制剂Ş有效的60天。其他制剂是乳化浓缩物（EC）和粉剂。粉尘制剂是难以适用于顶部和盒，其中，侦察蜜蜂花大部分时间进行评估散步两侧。虽然欧盟的配方是常见的，容易申请，他们仍然有效只有两个星期。不建议使用的天然产物除虫菊，因为它是感光，仅几天有效。苄氯菊酯是首选的杀虫剂，因为是它相对无毒鸟类和人类。它不会长期存在于​​环境，是防护剂对蜜蜂和持续时间比自然衍生化工更长的时间。由于这些原因菊酯喷雾制剂建议。 试图保持AHB出来的领域未能整个美洲。在佛罗里达州捕获港口附近，延迟AHB的到来，而到2007年，他们在全州南部建立的。由AHB为主的国家，有小号意味着放弃使出学习如何管理AHB。在管理AHB不违法的地区，即进入陷阱箱蜜蜂可以很容易地被删除，养蜂利用。它们可以被重新定位到管理一个养蜂场或提供给当地的养蜂人。通过去除殖民盒结合当地的养蜂人，协助项目可以受益双方当事人。养蜂人得到免费蜜蜂和生物学家少了一个任务与关心自己。 这推挽方法，通过AHB大大降低了谷仓猫头鹰巢箱定植。 AHB的防御性质和巢箱取出群的难度，使预防的协议至关重要。该协议可以用于任何空腔嵌套鸟以及空腔嵌套哺乳动物。它可能被证明是在最近推出的亚洲蜜蜂许多濒危鹦鹉整个加勒比地区，南美，非洲，甚至澳大利亚复苏的重要工具保护（东方蜜蜂 ）正在迅速蔓延。

由于意外引入二十多个非洲蜜蜂皇后到巴西圣保罗，1957年1，这些昆虫已遍布南美洲，中美洲和成美国南部，盗用和更换较少很好地适应欧洲蜜蜂（ EHB）的野生种群。这些混合非洲化蜜蜂（AHB）有把他们与两个人类和野生动物冲突的特征。他们表现出高度的防御行为，繁殖速度非常快，并且是在哪里，他们营造一个家1,2选择性较差。 虽然巢箱已经长期用于通过鸟类由空腔筑巢的鸟类3,4-以提高养殖，它们的使用已被证明是用于增强在热带5,6威胁和濒危鹦鹉的繁殖特别有效。不幸的是，非洲化蜜蜂往往选择这些相同的巢箱 - 有时甚至杀害乘员秒。此外，这些AHB会对生物学家显著的威胁时，他们正在进行研究的巢。在AHB已经建立南美地区，一些鹦鹉保护项目已经看到AHB 7-10占领了他们的人工巢箱。比赛巢箱与蜜蜂很普遍，而且不仅限于鹦鹉。无论是在新的和旧世界其他许多鸟类都受到影响11-15。如果没有努力减轻这些侵入AHB的威胁，近期鹦鹉保护的成功可能会丢失。 从蜜蜂的竞争管理的现行做法包括繁殖季节8，或化学品到产蛋箱15外侧的申请后，立即重复使用，高剂量的氯菊酯7，手动删除蜂拥而来，成交巢入口的。劳动参与和危险生物学家使这些战略不理想。一个preventio发展N协议是必要的。 蜜蜂菌落通过称为蜂拥，当老女王叶工蜂16达四分之三菌落的方法繁殖。非洲化蜜蜂"高繁殖率使他们更频繁地一窝蜂超过EHB（每年四到八倍）（谁蜂拥而上一次或每年两次）17。此外，AHB全年可，而不是在特定的季节一样蜂拥而上EHB。因此，AHB可以迅速占据最合适的筑巢地点在一个特定的区域。 可以利用蜜蜂找房子的行为来影响他们的选择巢址的。蜜蜂在群一小部分飞了出去，找到并检查新的潜在筑巢地点。几个潜在的筑巢地点都为球探同时。这些侦察蜜蜂返回到群和执行舞蹈指示位点的位置。越强的舞蹈是一个网站，更快的上海合作组织的数量UT蜜蜂在该网站18积聚。因此，蜜蜂更优质的网站跳舞更​​强，相比于那些平庸或很差的网站。一旦达到法定人数阈值时，群起飞，飞往新选定的主站点18。 亚致死剂量接触杀虫剂氯菊酯的阻碍球探蜜蜂来定位，并降低其活性水平19,20的能力。在第一次内部巢箱菊酯的应用，应当防护剂侦察蜂21，和加班，化学降解，其包装盒的评估过程中暴露于氯菊酯侦察蜜蜂，应该呈现不太能够进行强有力的招聘舞蹈，减少的金额童军招募蜜蜂的网站。一个更适合的（理想腔容积，小入口位于底部孔，信息素引诱）备用巢址是在蜜蜂陷阱盒的形式提供。这个优秀的陷阱网站应达到法定人数快比杀虫剂处理过的那只箱子领先群选择在鸟箱陷阱框。 作为一个例子，我们描述了一个实验通过AHB在南佛罗里达州蔗田减少谷仓猫头鹰人工巢箱定植。我们的目标是测试使用推拉方法，将来自与杀虫剂处理过的猫头鹰盒子"推"离蜜蜂和"拉"他们走向性信息素诱捕陷阱群的有效性。

<table border="0" cellpadding="0" cellspacing="0" width="652">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Citral (ACROS Organics)</td>
			<td style="width:117px;">Fisher Scientific</td>
			<td style="width:119px;">AC11044-1000</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Geraniol (ACROS Organics)</td>
			<td style="width:117px;">Fisher Scientific</td>
			<td>AC41090-1000</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PermaCap CS</td>
			<td style="width:117px;">BASF</td>
			<td style="width:119px;"></td>
			<td>controlled release permethrin</td>
		</tr>
	</tbody>
</table>

a push-pull protocol to reduce colonization of bird nest boxes by honey bees

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. These bees select cavities that are suitable nest sites for birds, resulting in competition. The difficulty of removing bees and their defensive behavior makes a prevention protocol necessary. Here, we describe a push-pull integrated pest management protocol to deter bees from inhabiting bird boxes by applying a bird safe insecticide, permethrin, to repel bees from nest boxes, while simultaneously attracting them to pheromone-baited swarm traps. Shown here is an example experiment using Barn Owl nest boxes. This protocol successfully reduced colonization of Barn Owl nest boxes by Africanized honey bees. This protocol is flexible, allowing adjustments to accommodate a wide range of bird species and habitats. This protocol could benefit conservation efforts where AHB are located.

在巴西进行的一项以前的小规模的试点推挽研究表明群陷阱是有效的，减少的鹦鹉巢箱22蜜蜂占领。要确定是否推拉方式可以在更大的范围内有效的通过AHB降低燕窝盒定植，我们进行了在佛罗里达州南部，美国甘蔗地实地研究。六十谷仓猫头鹰巢箱被安装，每一个用仅有15米处一大群陷阱配对。每群陷阱收到了信息素引诱。奇数猫头鹰盒子是用氯菊酯微胶囊喷雾杀虫剂处理过的每60天及偶数巢箱都留给未经处理的控制。巢箱和群陷阱被检查每周1）侦察蜜蜂存在或不存在和2）蜂群。 盒和陷阱进行了自2014年3至7月2015年的77成群抓监测，75例在群陷阱（&lt;STRONG&gt;图1）。只有两个猫头鹰盒这个实验过程中定植;两者都是未经处理的。我们让蜜蜂留在陷阱盒最少两个星期之前，我们搬到他们离开一个养蜂场4公里。为了减少蜜蜂潜逃（离开陷阱箱）的机会，我们让他们留在陷阱箱附加的两个星期前，他们被转移到一个新的蜂巢框之前。蜂巢和碎片刮出陷阱盒子，它在实验网站，并重新安装一个新的诱惑放在里面。显著更多的蜜蜂选择群陷阱未处理过的猫头鹰盒（X 2（DF = 1）= 29.2，P &lt;0.0001），治疗猫头鹰盒（X 2（DF = 1）= 36.1，P &lt;0.0001）由Pearson的智确定方检验耶茨&#39;连续性校正。有几个陷阱箱蜜蜂定植不止一次，12陷阱盒被蜜蜂占领了两次，五盒分别占据三次，两个箱子分别占据<html>四次。进行统计分析，每陷阱箱占用只计算一次。在一个巢址以前的蜂群的气味可以吸引新的殖民地23所以以前可能占用陷阱盒比无人居住盒更具吸引力。虽然一些治疗猫头鹰箱被蜜蜂球探，没有成群搬进这些盒子。在附近的控制点（含30谷仓猫头鹰箱）蜜蜂殖民箱63％。我们发现，使用群陷阱大大减少蜂群占据猫头鹰盒的数量。没有治疗的巢箱定植，表明氯菊酯是从殖民巢箱保持AHB有效。然而，仅仅群陷阱是有效的，因此菊酯的应用只能在关键的时间段（ 即蜂群赛季之前，为了鸟类繁殖季节），而不是将全年使用。 "SRC ="/文件/ ftp_upload / 53950 / 53950fig1.jpg"/&gt; 图1.由蜜蜂箱定植数，人数，在佛罗里达州南部的甘蔗地定植无论是陷阱盒或未经处理的谷仓猫头鹰巢箱超过17个月的时间周期（2014年3月到七2015）非洲化蜜蜂成群的。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53950/53950fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees at JoVE.com

A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees

侵入非洲化蜜蜂防止燕窝盒定植是鸟巢现场有限的鸟类保护工作的重要。我们提供了一个综合虫害管理办法"推"蜜蜂远离巢箱用防护剂杀虫剂，氯菊酯，和"拉"他们走向性信息素诱捕陷阱群。

推挽议定书蜜蜂会影响家禽的殖民化巢箱

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. ...

a-push-pull-protocol-to-reduce-colonization-bird-nest-boxes-honey

Nanyang Technological University

Research

JoVE Journal

Environment

8.6K 次观看.  University of Florida. 侵入非洲化蜜蜂防止燕窝盒定植是鸟巢现场有限的鸟类保护工作的重要。我们提供了一个综合虫害管理办法"推"蜜蜂远离巢箱用防护剂杀虫剂，氯菊酯，和"拉"他们走向性信息素诱捕陷阱群。 

视频: A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of natural proteins have a molecular mass between 40 - 200 kDa1,2, a robust and high-throughput method with a nanometer resolution capability is needed. Negative staining (NS) electron microscopy (EM) is an easy, rapid, and qualitative approach which has frequently been used in research laboratories to examine protein structure and protein-protein interactions. Unfortunately, conventional NS protocols often generate structural artifacts on proteins, especially with lipoproteins that usually form presenting rouleaux artifacts. By using images of lipoproteins from cryo-electron microscopy (cryo-EM) as a standard, the key parameters in NS specimen preparation conditions were recently screened and reported as the optimized NS protocol (OpNS), a modified conventional NS protocol 3 . Artifacts like rouleaux can be greatly limited by OpNS, additionally providing high contrast along with reasonably high&#8208;resolution (near 1 nm) images of small and asymmetric proteins. These high-resolution and high contrast images are even favorable for an individual protein (a single object, no average) 3D reconstruction, such as a 160 kDa antibody, through the method of electron tomography4,5. Moreover, OpNS can be a high&#8208;throughput tool to examine hundreds of samples of small proteins. For example, the previously published mechanism of 53 kDa cholesteryl ester transfer protein (CETP) involved the screening and imaging of hundreds of samples 6. Considering cryo-EM rarely successfully images proteins less than 200 kDa has yet to publish any study involving screening over one hundred sample conditions, it is fair to call OpNS a high-throughput method for studying small proteins. Hopefully the OpNS protocol presented here can be a useful tool to push the boundaries of EM and accelerate EM studies into small protein structure, dynamics and mechanisms.

More than half of proteins are small proteins (molecular mass &#60;200 kDa) that are challenging for both electron microscope imaging and three-dimensional reconstructions. Optimized negative staining is a robust and high-throughput protocol to obtain high contrast and relatively high resolution (~1 nm) images of small asymmetric proteins or complexes under different physiological conditions.

优化负染:通过电子显微镜检查小和不对称蛋白结构的高通量协议

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of ...

科研

环境科学

Experimental Protocol for Manipulating Plant-induced Soil Heterogeneity

Coexistence theory has often treated environmental heterogeneity as being independent of the community composition; however biotic feedbacks such as plant-soil feedbacks (PSF) have large effects on plant performance, and create environmental heterogeneity that depends on the community composition. Understanding the importance of PSF for plant community assembly necessitates understanding of the role of heterogeneity in PSF, in addition to mean PSF effects. Here, we describe a protocol for manipulating plant-induced soil heterogeneity. Two example experiments are presented: (1) a field experiment with a 6-patch grid of soils to measure plant population responses and (2) a greenhouse experiment with 2-patch soils to measure individual plant responses. Soils can be collected from the zone of root influence (soils from the rhizosphere and directly adjacent to the rhizosphere) of plants in the field from conspecific and heterospecific plant species. Replicate collections are used to avoid pseudoreplicating soil samples. These soils are then placed into separate patches for heterogeneous treatments or mixed for a homogenized treatment. Care should be taken to ensure that heterogeneous and homogenized treatments experience the same degree of soil disturbance. Plants can then be placed in these soil treatments to determine the effect of plant-induced soil heterogeneity on plant performance. We demonstrate that plant-induced heterogeneity results in different outcomes than predicted by traditional coexistence models, perhaps because of the dynamic nature of these feedbacks. Theory that incorporates environmental heterogeneity influenced by the assembling community and additional empirical work is needed to determine when heterogeneity intrinsic to the assembling community will result in different assembly outcomes compared with heterogeneity extrinsic to the community composition.

Understanding the role of environmental heterogeneity in species coexistence has typically focused on types of heterogeneity that are extrinsic to the community&#8217;s species composition. We provide novel detailed methods for creating soil heterogeneity treatments using soils subject to plant-soil feedback conditioning, or heterogeneity intrinsic to the community composition.

用于操纵植物诱导土壤异质性的实验协议

Coexistence theory has often treated environmental heterogeneity as being independent of the community composition; however biotic feedbacks such as ...

A Protocol for Conducting Rainfall Simulation to Study Soil Runoff

Rainfall is a driving force for the transport of environmental contaminants from agricultural soils to surficial water bodies via surface runoff. The objective of this study was to characterize the effects of antecedent soil moisture content on the fate and transport of surface applied commercial urea, a common form of nitrogen (N) fertilizer, following a rainfall event that occurs within 24&#160;hr&#160;after fertilizer application. Although urea is assumed to be readily hydrolyzed to ammonium and therefore not often available for transport, recent studies suggest that urea can be transported from agricultural soils to coastal waters where it is implicated in harmful algal blooms. A rainfall simulator was used to apply a consistent rate of uniform rainfall across packed soil boxes that had been prewetted to different soil moisture contents. By controlling rainfall and soil physical characteristics, the effects of antecedent soil moisture on urea loss were isolated. Wetter soils exhibited shorter time from rainfall initiation to runoff initiation, greater total volume of runoff, higher urea concentrations in runoff, and greater mass loadings of urea in runoff. These results also demonstrate the importance of controlling for antecedent soil moisture content in studies designed to isolate other variables, such as soil physical or chemical characteristics, slope, soil cover, management, or rainfall characteristics. Because rainfall simulators are designed to deliver raindrops of similar size and velocity as natural rainfall, studies conducted under a standardized protocol can yield valuable data that, in turn, can be used to develop models for predicting the fate and transport of pollutants in runoff.

A rainfall simulator was used to apply a consistent rate of uniform rainfall to packed soil boxes in a study of the fate and transport of urea, a nonpoint source environmental contaminant. Under uniform soil and rainfall conditions, antecedent soil moisture content exerted strong control over urea loss in surface runoff.

一个协议，用于开展降雨模拟研究土壤径流

Rainfall is a driving force for the transport of environmental contaminants from agricultural soils to surficial water bodies via surface runoff. The ...

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 ...

Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level

The presence of pesticides in the beekeeping environment is one of the most serious problems that impacts the life of a honey bee. Pesticides can be brought back to the beehive after the bees have foraged on flowers that have been sprayed with pesticides. Pesticide contaminated food can be exchanged between workers which then feed larvae and therefore can potentially affect the development of honey bees. Thus, residual pesticides in the environment can become a chronic damaging factor to honey bee populations and gradually lead to colony collapse. In the presented protocol, honey bee feeding methods are described and applied to either an individual honey bee or to a colony. Here, the insect growth regulator (IGR) pyriproxyfen (PPN), which is widely used to control pest insects and is harmful to the development of honey bee larvae and pupae, is used as the pesticide. The presenting procedure can be applied to other potentially harmful chemicals or honeybee pathogens for further studies.

Herein we present a method to feed pesticide contaminated food to both an individual honey bee and a beehive colony. The procedure evaluates the pesticide effect on individual honey bees by in vivo feeding of basic larval diet and also on the natural condition of beehive colony.

从个体到群体度评价环境化学物质对蜜蜂发展的影响

The presence of pesticides in the beekeeping environment is one of the most serious problems that impacts the life of a honey bee. Pesticides can be ...

Video Tracking Protocol to Screen Deterrent Chemistries for Honey Bees

The European honey bee, Apis mellifera L., is an economically and agriculturally important pollinator that generates billions of dollars annually. Honey bee colony numbers have been declining in the United States and many European countries since 1947. A number of factors play a role in this decline, including the unintentional exposure of honey bees to pesticides. The development of new methods and regulations are warranted to reduce pesticide exposures to these pollinators. One approach is the use of repellent chemistries that deter honey bees from a recently pesticide-treated crop. Here, we describe a protocol to discern the deterrence of honey bees exposed to select repellent chemistries. Honey bee foragers are collected and starved overnight in an incubator 15 h prior to testing. Individual honey bees are placed into Petri dishes that have either a sugar-agarose cube (control treatment) or sugar-agarose-compound cube (repellent treatment) placed into the middle of the dish. The Petri dish serves as the arena that is placed under a camera in a light box to record the honey bee locomotor activities using video tracking software. A total of 8 control and 8 repellent treatments were analyzed for a 10 min period with each treatment was duplicated with new honey bees. Here, we demonstrate that honey bees are deterred from the sugar-agarose cubes with a compound treatment whereas honey bees are attracted to the sugar-agarose cubes without an added compound.

The loss of honey bee colonies presents a challenge to crop pollination services. Current pollinator protection practices warrant an alternative approach to minimize the contact of honey bees to harmful pesticides using repellent chemistries. Here, we provide detailed methods for a visual tracking protocol to screen deterrents for bees.

用于蜂蜜蜜蜂筛选化学成分的视频跟踪协议

The European honey bee, Apis mellifera L., is an economically and agriculturally important pollinator that generates billions of dollars annually. Honey ...

Experimental Protocol for Detecting Mitochondrial Function in Hepatocytes Exposed to Organochlorine Pesticides

This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by environmental organochlorine pesticides (OCPs) in hepatocytes. HepG2 cells were exposed to &#946;-hexachlorocyclohexane (&#946;-HCH) for 24 h at an equivalent dose of internal exposure in general population. Ultrastructure in hepatocytes was examined by transmission electron microscopy (TEM) to show the damage of mitochondria. Mitochondrial function was further evaluated by mitochondrial fluorescence intensity, adenosine 5'-triphosphate (ATP) levels, oxygen consumption rate (OCR) and mitochondrial membrane potential (MMP) in HepG2 cells incubated with &#946;-HCH. The mitochondria fluorescence intensity after stained by mitochondrial green fluorescent probe was observed with a fluorescence microscopy. The luciferin-luciferase reaction was used to determine ATP levels. The MMP was detected by the cationic dye JC-1 and analyzed under flow cytometry. OCR was measured with an extracellular flux analyzer. In summary, these protocols were used in detecting mitochondrial function in hepatocytes with to investigate mitochondria damages.

Understanding the influence of environmental organochlorine pesticides (OCPs) on mitochondrial function in hepatocytes is important in exploring the mechanism of OCPs causing metabolic disorders. This paper presents detailed methods on detecting hepatic mitochondrial function.

检测暴露于有机氯农药的肝细胞线粒体功能的实验方案

This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by ...

In Vitro Rearing of Solitary Bees: A Tool for Assessing Larval Risk Factors

Although solitary bees provide crucial pollination services for wild and managed crops, this species-rich group has been largely overlooked in pesticide regulation studies. The risk of exposure to fungicide residues is likely to be especially high if the spray occurs on, or near host plants while the bees are collecting pollen to provision their nests. For species of Osmia that consume pollen from a select group of plants (oligolecty), the inability to use pollen from non-host plants can increase their risk factor for fungicide-related toxicity. This manuscript describes protocols used to successfully rear oligolectic mason bees, Osmia ribifloris sensu lato, from egg to prepupal stage within cell culture plates under standardized laboratory conditions. The in vitro-reared bees are subsequently used to investigate the effects of fungicide exposure and pollen source on bee fitness. Based on a 2 &#215; 2 fully crossed factorial design, the experiment examines the main and interactive effects of fungicide exposure and pollen source on larval fitness, quantified by prepupal biomass, larval developmental time, and survivorship. A major advantage of this technique is that using in vitro-reared bees reduces natural background variability and allows the simultaneous manipulation of multiple experimental parameters. The described protocol presents a versatile tool for hypotheses testing involving the suite of factors affecting bee health. For conservation efforts to be met with significant, lasting success, such insights into the complex interplay of physiological and environmental factors driving bee declines will prove to be critical.

In Vitro Rearing of Solitary Bees: A Tool for Assessing Larval Risk Factors

Fungicide sprays on flowering plants may expose solitary bees to high concentrations of pollen-borne fungicide residues. Using laboratory-based experiments involving in vitro-reared bee larvae, this study investigates the interactive effects of consuming fungicide-treated pollen derived from host and non-host plants.

体外孤蜂饲养: 一种评估幼虫危险因素的工具

Although solitary bees provide crucial pollination services for wild and managed crops, this species-rich group has been largely overlooked in pesticide ...

Experimental Protocol for Examining Behavioral Response Profiles in Larval Fish: Application to the Neuro-stimulant Caffeine

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and toxicological studies. Using automated digital tracking platforms, recent efforts in neuropharmacology are leveraging larval fish locomotor behaviors to identify potential therapeutic targets for novel small molecules. Similar to these efforts, research in the environmental sciences and comparative pharmacology and toxicology is examining various behaviors of fish models as diagnostic tools in tiered evaluation of contaminants and real-time monitoring of surface waters for contaminant threats. Whereas the zebrafish is a popular larval fish model in the biomedical sciences, the fathead minnow is a common larval fish model in ecotoxicology. Unfortunately, fathead minnow larvae have received considerably less attention in behavioral studies. Here, we develop and demonstrate a behavioral profile protocol using caffeine as a model neurostimulant. Though photomotor responses of fathead minnows were occasionally affected by caffeine, zebrafish&#160;were markedly more sensitive for photomotor and locomotor endpoints, which responded at environmentally relevant levels. Future studies are needed to understand comparative behavioral sensitivity differences among fish with age and time of day, and to determine whether similar behavioral effects would occur in nature and be indicative of adverse outcomes at the individual or population levels of biological organization.

Here, we present a protocol to examine larval zebrafish and fathead minnow locomotor activities and photomotor responses (PMR) using an automated tracking software. When incorporated in common toxicology bioassays, analyses of these behaviors provide a diagnostic tool to examine chemical bioactivity. This protocol is described using caffeine, a model neurostimulant.

检测幼虫行为反应剖面的实验协议: 在神经刺激咖啡因中的应用

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and ...

Measuring Hypopharyngeal Gland Acinus Size in Honey Bee (Apis mellifera) Workers

The nurse hypopharyngeal glands produce the protein fraction of the worker and royal jelly that is fed to developing larvae and queens. These paired glands that are located in the head of the bee are highly sensitive to the quantity and quality of pollen and pollen substitutes that the nurse bee consumes. The glands get smaller when nurses are fed deficient diets and are large when they are fed complete diets. Because nurse hypopharyngeal gland size is a robust indicator of nurse nutrition, it is essential that those studying honey bee nutrition know how to measure these glands. Here, we provide detailed methods for dissecting, staining, imaging, and measuring nurse bee hypopharyngeal glands. We present comparisons of unstained and stained tissue and data that were used to study the impact of pollen on gland size. This method has been used to test how diet impacts hypopharyngeal gland size but has further use for understanding the role of these glands in hive health.

Measuring Hypopharyngeal Gland Acinus Size in Honey Bee Apis mellifera Workers

Hypopharyngeal gland acinus size is a robust measure of nurse honey bee nutrition. Here, we provide a detailed protocol for dissecting, staining, imaging, and measuring nurse bee hypopharyngeal gland acini.

蜂蜜蜂 (api 蜜蜂) 中下咽腺性腺泡大小的测定

The nurse hypopharyngeal glands produce the protein fraction of the worker and royal jelly that is fed to developing larvae and queens. These paired ...