Name: a push-pull protocol to reduce colonization of bird nest boxes by honey bees
Uploaded: 2016-09-04T13:00:00.000+00:00
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Description: Watch this Scientific Journal Video about A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees at JoVE.com

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><tbody><tr><td>Citral (ACROS Organics)</td><td>Fisher Scientific</td><td>AC11044-1000</td><td></td></tr><tr><td>Geraniol (ACROS Organics)</td><td>Fisher Scientific</td><td>AC41090-1000</td><td></td></tr><tr><td>PermaCap CS</td><td>BASF</td><td></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

Empirical, Metagenomic, and Computational Techniques Illuminate the Mechanisms by which Fungicides Compromise Bee Health

Growers often use fungicide sprays during bloom to protect crops against disease, which exposes bees to fungicide residues. Although considered &#34;bee-safe,&#34; there is mounting evidence that fungicide residues in pollen are associated with bee declines (for both honey and bumble bee species). While the mechanisms remain relatively unknown, researchers have speculated that bee-microbe symbioses are involved. Microbes play a pivotal role in the preservation and/or processing of pollen, which serves as nutrition for larval bees. By altering the microbial community, it is likely that fungicides disrupt these microbe-mediated services, and thereby compromise bee health. This manuscript describes the protocols used to investigate the indirect mechanism(s) by which fungicides may be causing colony decline. Cage experiments exposing bees to fungicide-treated flowers have already provided the first evidence that fungicides cause profound colony losses in a native bumble bee (Bombus impatiens). Using field-relevant doses of fungicides, a series of experiments have been developed to provide a finer description of microbial community dynamics of fungicide-exposed pollen. Shifts in the structural composition of fungal and bacterial assemblages within the pollen microbiome are investigated by next-generation sequencing and metagenomic analysis. Experiments developed herein have been designed to provide a mechanistic understanding of how fungicides affect the microbiome of pollen-provisions. Ultimately, these findings should shed light on the indirect pathway through which fungicides may be causing colony declines.

Microbial consortia within bumble bee hives enrich and preserve pollen for bee larvae. Using next generation sequencing, along with laboratory and field-based experiments, this manuscript describes protocols used to test the hypothesis that fungicide residues alter the pollen microbiome, and colony demographics, ultimately leading to colony loss.

经验、基因和计算技术阐明了杀菌剂危害蜜蜂健康的机制

Growers often use fungicide sprays during bloom to protect crops against disease, which exposes bees to fungicide residues. Although considered ...

科研

环境科学

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 ...

A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). There are several behavioral mechanisms through which an insect learns salient associations and relates them to these events. It is important to understand this behavioral plasticity for programs aimed toward assisting insects that are beneficial for agriculture. This understanding can also be used for discovering solutions to biomedical and agricultural problems created by insects that act as disease vectors and pests. The Proboscis Extension Response (PER) conditioning protocol was developed for honey bees (Apis mellifera) over 50 years ago to study how they perceive and learn about floral odors, which signal the nectar and pollen resources a colony needs for survival. The PER procedure provides a robust and easy-to-employ framework for studying several different ecologically relevant mechanisms of behavioral plasticity. It is easily adaptable for use with several other insect species and other behavioral reflexes. These protocols can be readily employed in conjunction with various means for monitoring neural activity in the CNS via electrophysiology or bioimaging, or for manipulating targeted neuromodulatory pathways. It is a robust assay for rapidly detecting sub-lethal effects on behavior caused by environmental stressors, toxins or pesticides.
We show how the PER protocol is straightforward to implement using two procedures. One is suitable as a laboratory exercise for students or for quick assays of the effect of an experimental treatment. The other provides more thorough control of variables, which is important for studies of behavioral conditioning. We show how several measures for the behavioral response ranging from binary yes/no to more continuous variable like latency and duration of proboscis extension can be used to test hypotheses. And, we discuss some pitfalls that researchers commonly encounter when they use the procedure for the first time.

The Proboscis Extension Response (PER) conditioning protocol, developed for the honey bee (Apis mellifera), provides an ecologically-relevant and easily quantifiable means for studying several different mechanisms of learning in many insect species.

长鼻扩展响应协议为侦查行为可塑性的昆虫：应用基础，生物医学和农业研究

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). ...

神经科学

Preparation of Single-cohort Colonies and Hormone Treatment of Worker Honeybees to Analyze Physiology Associated with Role and/or Endocrine System

Honeybee workers are engaged in various tasks related to maintaining colony activity. The tasks of the workers change according to their age (age-related division of labor). Young workers are engaged in nursing the brood (nurse bees), while older workers are engaged in foraging for nectar and pollen (foragers). The physiology of the workers changes in association with this role shift. For example, the main function of the hypopharyngeal glands (HPGs) changes from the secretion of major royal jelly proteins (MRJPs) to the secretion of carbohydrate-metabolizing enzymes. Because worker tasks change as the workers age in typical colonies, it is difficult to discriminate the physiological changes that occur with aging from those that occur with the role shift. To study the physiological changes in worker tissues, including the HPGs, in association with the role shift, it would be useful to manipulate the honeybee colony population by preparing single-cohort colonies in which workers of almost the same age perform different tasks. Here we describe a detailed protocol for preparing single-cohort colonies for this analysis. Six to eight days after single-cohort colony preparation, precocious foragers that perform foraging tasks earlier than usual appear in the colony. Representative results indicated role-associated changes in HPG gene expression, suggesting role-associated HPG function. In addition to manipulating the colony population, analysis of the endocrine system is important for investigating role-associated physiology. Here, we also describe a detailed protocol for treating workers with 20-hydroxyecdysone (20E), an active form of ecdysone, and methoprene, a juvenile hormone analogue. The survival rate of treated bees was sufficient to examine gene expression in the HPGs. Gene expression changes were observed in response to 20E- and/or methoprene-treatment, suggesting that hormone treatments induce physiological changes of the HPGs. The protocol for hormone treatment described here is appropriate for examining hormonal effects on worker physiology.

Here we describe our detailed protocol for the preparation of single-cohort honeybee colonies&#160;&#8211; a useful tool for analyzing the role-associated worker physiology. We also describe detailed protocols for treating workers with juvenile hormone and ecdysone to evaluate the involvement of these hormones in the regulation of worker behavior and/or physiology.

单队列殖民地和工蜂的激素治疗的准备来分析生理学与关联作用和/或内分泌系统

Honeybee workers are engaged in various tasks related to maintaining colony activity. The tasks of the workers change according to their age (age-related ...

Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera

Honey bees demonstrate astonishing learning abilities and advanced social behavior and communication. In addition, their brain is small, easy to visualize and to study. Therefore, bees have long been a favored model amongst neurobiologists and neuroethologists for studying the neural basis of social and natural behavior. It is important, however, that the experimental techniques used to study bees do not interfere with the behaviors being studied. Because of this, it has been necessary to develop a range of techniques for pharmacological manipulation of honey bees. In this paper we demonstrate methods for treating restrained or free-flying honey bees with a wide range of pharmacological agents. These include both noninvasive methods such as oral and topical treatments, as well as more invasive methods that allow for precise drug delivery in either systemic or localized fashion. Finally, we discuss the advantages and disadvantages of each method and&#160;describe common hurdles and how to best overcome them. We conclude with a discussion on the importance of adapting the experimental method to the biological questions rather than the other way around.

Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera

This manuscript describes several protocols for administering pharmacological agents to honey bees, including simple noninvasive methods for free-flying bees, as well as more invasive variants that allow precise localized treatment of restrained bees.

约束和自由飞行，蜜蜂的神经药理学操纵，<em&gt;蜜蜂</em

Honey bees demonstrate astonishing learning abilities and advanced social behavior and communication. In addition, their brain is small, easy to visualize ...

Monitoring Colony-level Effects of Sublethal Pesticide Exposure on Honey Bees

The effects of sublethal pesticide exposure to honey bee colonies may be significant but difficult to detect in the field using standard visual assessment methods. Here we describe methods to measure the quantities of adult bees, brood, and food resources by weighing hives and hive parts, by photographing frames, and by installing hives on scales and with internal sensors. Data from these periodic evaluations are then combined with running average and daily detrended data on hive weight and internal hive temperature. The resulting datasets have been used to detect colony-level effects of imidacloprid applied in a sugar syrup as low as 5 parts per billion. The methods are objective, require little training, and provide permanent records in the form of sensor output and photographs.

Sublethal doses of pesticides may affect colonies in ways that are difficult to detect using only periodic or visual methods. Methods to measure total adult bee mass, brood, and food resources by weighing hives and hive parts, photographing frames, and installing sensors, are provided. Data analysis is also addressed.

蜜蜂致死农药暴露对蜂群水平的影响

The effects of sublethal pesticide exposure to honey bee colonies may be significant but difficult to detect in the field using standard visual assessment ...

生物学

A 3D Printed Pollen Trap for Bumble Bee (Bombus) Hive Entrances

To verify the plant sources from which bumble bees forage for pollen, individuals must be collected to remove their corbicular pollen loads for analysis. This has traditionally been done by netting foragers at nest entrances or on flowers, chilling the bees on ice, and then removing the pollen loads from the corbiculae with forceps or a brush. This method is time and labor intensive, may alter normal foraging behavior, and can result in stinging incidents for the worker performing the task. Pollen traps, such as those used on honey bee hives, collect pollen by dislodging corbicular pollen loads from the legs of workers as they pass through screens at the nest entrance. Traps can remove a large quantity of pollen from returning forager bees with minimal labor, yet to date no such trap is available for use with bumble bee colonies. Workers within a bumble bee colony can vary in size making size selection of entrances difficult to adapt this mechanism to commercially reared bumble bee hives. Using 3D printing design programs, we created a pollen trap that successfully removes the corbicular pollen loads from the legs of returning bumble bee foragers. This method significantly reduces the amount of time required by researchers to collect pollen from bumble bee foragers returning to the colony. We present the design, results of pollen removal efficiency tests, and suggest areas of modifications for investigators to adapt traps to a variety of bumble bee species or nest box designs.

A 3D Printed Pollen Trap for Bumble Bee Bombus Hive Entrances

We present a non-lethal and automated mechanism to collect pollen from bumble bee (Bombus) workers returning to a hive. Instructions for producing, preparing, installing and using the devices are included. By using 3D-printed objects, modification to the design was timely, efficient and allowed for quick turnaround for testing.

用于大黄蜂（Bombus）蜂巢入口的3D打印花粉陷阱

To verify the plant sources from which bumble bees forage for pollen, individuals must be collected to remove their corbicular pollen loads for analysis.  ...

Collection and Identification of Pollen from Honey Bee Colonies

Researchers often collect and analyze corbicular pollen from honey bees to identify the plant sources on which they forage for pollen or to estimate pesticide exposure of bees via pollen. Described herein is an effective pollen-trapping method for collecting corbicular pollen from honey bees returning to their hives. This collection method results in large quantities of corbicular pollen that can be used for research purposes. Honey bees collect pollen from many plant species, but typically visit one species during each collection trip. Therefore, each corbicular pollen pellet predominantly represents one plant species, and each pollen pellet can be described by color. This allows the sorting of samples of corbicular pollen by color to segregate plant sources. Researchers can further classify corbicular pollen by analyzing the morphology of acetolyzed pollen grains for taxonomic identification. These methods are commonly used in studies related to pollinators such as pollination efficiency, pollinator foraging dynamics, diet quality, and diversity. Detailed methodologies are presented for collecting corbicular pollen using pollen traps, sorting pollen by color, and acetolyzing pollen grains. Also presented are results pertaining to the frequency of pellet colors and taxa of corbicular pollen collected from honey bees in five different cropping systems.

We describe methods for collecting corbicular pollen from honey bees as well as protocols for color sorting, acetolysis, and microscope slide preparation of pollen for taxonomic identification. In addition, we present pellet color and taxonomic diversity of corbicular pollen collected from five cropping systems using pollen traps.

蜜蜂群落花粉的采集和鉴定

Researchers often collect and analyze corbicular pollen from honey bees to identify the plant sources on which they forage for pollen or to estimate ...

Assessing Agrochemical Risk to Mated Honey Bee Queens

Current risk assessment strategies for honey bees rely heavily upon laboratory tests performed on adult or immature worker bees, but these methods may not accurately capture the effects of agrochemical exposure on honey bee queens. As the sole producer of fertilized eggs inside a honeybee colony, the queen is arguably the most important single member of a functioning colony unit. Therefore, understanding how agrochemicals affect queen health and productivity should be considered a critical aspect of pesticide risk assessment. Here, an adapted method is presented to expose honey bee queens and worker queen attendants to agrochemical stressors administered through a worker diet, followed by tracking egg production in the laboratory and assessing first instar eclosion using a specialized cage, referred to as a Queen Monitoring Cage. To illustrate the method&#39;s intended use, results of an experiment in which worker queen attendants were fed diet containing sublethal doses of imidacloprid and effects on queens were monitored are described.

This protocol was developed to enhance the understanding of how agrochemicals affect honey bee (Apis mellifera) reproduction by establishing methods to expose honey bee queens and their worker caretakers to agrochemicals in a controlled, laboratory setting and carefully monitoring their relevant responses.

评估交配蜜蜂女王的农用化学品风险

Current risk assessment strategies for honey bees rely heavily upon laboratory tests performed on adult or immature worker bees, but these methods may not ...