密歇根州立大学的几位技术员和研究生帮助开发、提炼和评估了多年来的 DD 陷阱设计, 包括安德烈 Anulewicz、罗伯特. 麦克唐纳和内森 Siegert。我们感谢詹姆斯 Wieferich 和杰里米·洛厄尔 (密歇根州立大学) 协助制定 DD 安装说明。詹姆斯 Wieferich 和莫莉 Robinett (密歇根州立大学) 审查了这份手稿的早期草稿, 我们赞赏他们的建议。约瑟夫弗兰切塞和戴蒙骗子 (USDA) 慷慨地分享了他们对 EAB 对颜色和寄主挥发物的反应的观察。为 DD 陷阱开发和评估提供资金的是由美国农业部森林服务局、东北地区、森林健康保护基金资助的。

1. 准备绿色和紫色的面板 <ol> <li> 获取绿色和紫色的波纹塑料板 (120 厘米和 #215; 60 厘米), 用于从商业分销商那里 EAB 诱捕害虫管理用品。使用盒式刀具或实用刀在每个面板上分两条折线, 通过沿垂直波纹的塑料部分切割, 从面板的两个短边40厘米。这使得面板折叠成一个3棱棱镜 (每张脸将是 40 x 60 厘米)。面板通常是分销商的得分, 以方便折叠, 所以这一步可能是不必要的. </li>
	<li> 使用 and #189 在每个面板中钻取两对孔; 如图1所示, 英寸 (1.3 厘米) 位 。钻洞4英寸 (10 厘米) 从面板的顶部和4英寸两侧的左边的褶皱。从面板顶部向下钻取19英寸 (48 厘米) 的第二对孔, 并直接位于顶部孔的下方 ( 即 , 左侧折叠两侧的4英寸). </li>
</ol> 2。准备 PVC 管道 <ol> <li> 从商业硬件商店或其他供应商处获取每个陷阱的计划40聚氯乙烯 (pvc) 管道、3米长和10厘米直径的一节。将四孔钻入 PVC 管道, 如 图 2 所示。使用 #189; 英寸 (1.3 厘米) 钻头, 从 PVC 管顶部钻出一对4英寸 (10 厘米) 的孔, 并将3和 #189; 英寸 (8.9 厘米) 分开。水平对齐孔. </li>
	<li> 钻取第二对孔, 3 和 #189; 英寸分开, 52 英寸 (1.3 米), 低于顶部的管道, 直接低于第一组孔. </li>
</ol> 3。安装 t post <ol> <li> 使用 post 磅在所需位置安装 t 开机自检。把柱子开得足够深, 这样法兰就在土壤表面下面, 但是没有必要再深入了。t 岗位将为 DD 陷阱提供大量支持, 即使在大风中也是如此。人导线或其他支持不是必要的。虽然 t 岗位不需要深深地被设置入土壤, 它是一个好想法检查地下公用事业或被埋没的管子在驾驶岗位之前入地面. </li>
</ol> 4。将 DD 陷阱 <ol> <li> 将紫色面板折叠成棱镜。如果面板是 pre-glued 的, 请确保粘着的一面在外面。用两个8英寸 (20 厘米) 的电缆连接, 将两个小的襟翼通过狭缝送进, 将面板两端固定在一起。重复使用绿色面板. </li>
	<li> 支持水平在地面上的 PVC 管道。将紫色的棱镜滑到 pvc 管上, 然后向下放到 pvc 管道的下一组孔中. </li>
	<li> 喂大, 2 英尺 (60 厘米) 长的电缆扎平, 通过在棱镜的左上孔, 然后 通过 在 PVC 管的一对孔, 最后通过相邻的孔在右侧的棱镜。确保棱镜紧贴在两个孔之间的 PVC 管的实心部分。关闭并拧紧电缆领带, 确保电缆领带不扭曲. </li>
	<li> 在棱镜的底部孔中通过第二个大的电缆扎接, 围绕着 管道和棱镜相邻侧的相应孔。关闭电缆领带和拧紧, 再次小心, 以确保电缆领带不拧紧之前, 它收紧. </li>
	<li> 将绿色棱镜滑到 pvc 管上, 并将其连接到 pvc 管道上的上一组孔中, 两个60厘米的电缆连接, 与前面的步骤相同。在上下棱镜之间将有一个21英寸 (54 厘米) 的间隙. </li>
</ol> 5。应用昆虫胶并设置陷阱 <ol> <li> 对不 pre-glued 的面板应用清除昆虫补漏白胶。甚至 pre-glued 板通常需要额外的胶水来捕获 EAB 甲虫。应用昆虫诱捕胶, 在地面上设置 PVC 管底座, 在 t 型柱上支撑上棱镜的内面. </li>
	<li> 使用乳胶 (或类似的) 手套, 舀起两到三把胶水, 涂抹在一个面板的表面, 确保面板的表面被彻底涂上。旋转 PVC 管道以访问两个棱镜上的所有三表面。胶水不需要应用到 PVC 管道. </li>
	<li> 拾起 PVC 管子, 垂直地保持它并且滑动低端在 t 岗位。避免触摸粘板. </li>
</ol> 6。诱饵的 DD 陷阱 <ol> <li> 作为 pre-glued 棱镜通常到达与一个小孔 (4 毫米) 靠近底部边缘的一个面板的棱镜, 使用4英寸 (10 厘米) 电缆领带, 将一袋 cis -3-hexenol 连接到顶部 (绿色) 棱镜的下边缘。如果一个孔不存在, 使用手持打孔或折刀切割一个小孔或缝进塑料, 然后使用小电缆领带附加诱饵. 
	注: 在诱饵中的 cis -3-hexenol 化合物在大约10周内慢慢释放。如果陷阱需要保持有效超过10周, 更换诱饵的需要. </li>
</ol> 7。检查 EAB 甲虫的陷阱 <ol> <li> 如果可能, 请在2-4 周间隔内检查陷阱, 以避免丢失捕获的甲虫, 这会偶尔掉落陷阱, 特别是在大雨发生时。检查下棱镜的所有三边为 EAB 甲虫。然后, 使用一次性乳胶手套, 升降机 PVC 管直上和关闭 t 柱和支柱内面的上部棱镜的 t 柱, 露出一个面板的两个棱镜. </li>
	<li> 使用镊子清除任何可疑的 EAB 甲虫, 并将它们放在一个密封塑料标本袋中. </li>
	<li> 旋转 PVC 管道以检查两个棱镜上的所有三面板是否有可疑的 EAB 甲虫, 然后将管道滑回 t 型柱上. 
	注意: 苍蝇, 点击甲虫和其他各种昆虫将不可避免地被捕获的陷阱。它需要敏锐的眼睛来发现一个 EAB 甲虫。在第一次检查陷阱时, 携带照片参考卡会很有帮助. </li>
	<li> 要确认物种识别, 将可疑甲虫送回实验室, 并将其浸泡在无毒的组织学清除剂中一两天, 以除去昆虫诱捕胶。甲虫可以在解剖显微镜下检查. </li>
</ol>

作者声明他们没有竞争的金融利益。

DD 陷阱的设计和放置利用了成年 EAB 甲虫对特定颜色色调和光照的吸引力。PVC 管顶部的绿色棱镜对雄性甲虫最有吸引力, 它们将生命周期用在灰叶上, 以及交配32、36、38、39。较低的紫色棱镜确保陷阱也吸引女性甲虫32。与雄性一样, 雌性甲虫在其生命跨度内以火山灰叶为食, 但成熟雌?...

翡翠灰钻 (EAB) (吉丁吉丁Fairmaire) (鞘翅目: 吉丁) 已经杀死了数亿的火山灰 (水曲柳) 树木, 因为它是首次确定在2002年在大底特律, 密歇根州大都市区和附近的温莎, 安大略省, 加拿大。无意中蔓延的灰树, 原木和木柴, 连同自然分散甲虫, 导致 EAB 建立在至少27州, 和两个加拿大省份的日期1。最近的报道表明, EAB 也入侵俄罗斯莫斯科, 在那里, 它是杀害景观灰树2,3, 引起更多的担心, 它的潜力蔓延到欧洲。北美灰种的 EAB 寄主偏好和抗性的物种间变异已被记录为4,5,6,78,9,但几乎所有的北美灰种都可能是合适的寄主。灰死亡率的灾难性水平已记录在密歇根州和俄亥俄州的地区10,11,12, 具有相关的生态和经济影响13,14, 15,16。
有效的方法来检测新的 EAB 虫害和监测低密度人口是管理这种侵入性害虫在城市, 居住区和森林环境的关键方面。早期发现为制定战略、保障资金和实施活动以减少 EAB 的影响提供了时间。例如, 市政官员和房主可以开始用全身杀虫剂在景观中处理有价值的火山灰, 然后在幼虫密度增加的情况下损害这些产品的功效17,18。同样, 查明新的侵扰和关于当地 EAB 分布的可靠信息, 使林农和财产所有人有机会实施木材销售、物种转换或其他活动, 以降低经济成本或生态灰死亡率的影响。
然而, 对低密度 EAB 种群的早期发现、划定和有效监测仍然很困难。目视调查, 以确定新出没的火山灰树是不可靠的, 因为火山灰很少表现出外部迹象或症状的 EAB 侵扰, 直到幼虫密度建设到中等甚至高级别4,19。检测低密度 EAB 的最有效方法涉及使用着灰检测树19,20,21,22。在春季或初夏, 灰树是着的, 去除树干周长周围的外树皮和韧皮部, 这就强调树木, 增加它们对成年 EAB 甲虫的吸引力。着树可以在秋季或冬季声带, 以确定 EAB 幼虫的存在和密度。虽然着灰树已在操作上用于 EAB 检测19、23、24、25, 但它们都有问题。剥皮着树可以是劳力密集型和寻找合适的树木, 剥可能是困难的, 特别是在城市或住宅区, 或当调查必须进行多年的19。
人工陷阱诱饵与 EAB 引诱消除了许多关注与使用着灰检测树。与其他重要的森林害虫, 如吉普赛蛾 (毒蛾L) 和一些 Scolytinae 树皮甲虫, 产生长距离的性别或聚集费洛蒙, 到目前为止, 没有有效的远距离信息素已经找到 EAB。短范围性信息素,顺式-内酯, 可促进交配26,27, 但在田间试验中,顺式内酯诱饵并没有始终如一地增加 EAB 吸引到人工陷阱28。成年甲虫依赖于由火山灰树叶、树皮和木材发出的挥发性化合物来识别它们的宿主树29、30、31并遇到潜在的伙伴。几个挥发性化合物已被评价用于引诱吸引成人 EABs 到人工陷阱27,32。目前, 在美国的 EAB 检测调查中使用的陷阱是诱饵含有cis-3-hexenol, 一个常见的绿叶挥发性产生的火山灰叶30,33。在前几年, 用于美国调查的 EAB 陷阱也被麦卢卡油, 这是从新西兰茶树提取 (Leptospermum 花马蹄和马蹄) 或菲比油, 巴西核桃树的提取物 (菲比鳐Mez。两者都含有几个萜, 也存在于灰树皮29中。然而, 这些天然油的供应不一致的问题限制了它们的使用。
除了寄主产生的挥发物, 成年 EAB 甲虫响应视觉刺激, 包括颜色和光20,32,34,35。早期的研究表明, EAB 成虫是相对敏捷的飞行物, 很少被各种火山灰挥发物引诱的黑色漏斗诱捕器捕获 (副总经理和 TMP, 未公布的数据)。其他陷阱设计, 如 cross-vane 陷阱, 被评估, 但 EAB 甲虫对黑暗的空间和阴影的反感限制了这些陷阱的有效性。
开发的三棱棱镜, 可以涂上明确的昆虫诱捕胶水35捕捉甲虫, 是一个重大的改进, 陷阱设计。成人 EABs 对颜色的吸引力在野外试验和实验室研究中也被广泛评价为 retinograph34。结果表明 EAB 甲虫一直被吸引到特定的绿色和紫色的色调32,36。在美国和加拿大的 EAB 调查活动中, 现在广泛使用彩色波纹塑料制作的棱镜阱。
因为 EAB 的成年人强烈地被光吸引, 甲虫更有可能在 open-grown 树上殖民着色树的20,21。美国 EAB 检测调查指南要求单个棱镜阱从沿道路生长的灰树中的 mid-canopy 分支或树林区域的边缘37暂停。理论上, 这将确保至少有一个棱镜的面板暴露在阳光下。然而, 在操作上, 棱镜陷阱可能会被头顶的树枝或相邻或附近的树木遮蔽。粘滞板表面经常被吹入叶子, 导致叶子坚持和遮蔽至少一部分的一个或多个面板。
双甲板 (DD) 陷阱被开发, 以整合多种视觉和嗅觉线索, 以提高 EAB 甲虫的吸引力。每个 DD 陷阱是由一个绿色和一个紫色波纹塑料棱镜附加到一个3米高的时间表40聚氯乙烯 (pvc) 管 (10 厘米直径), 这是支持滑动 PVC 管在 t 柱。使用绿色和紫色棱镜设计, 以吸引两性的 EAB 甲虫32,36,38,39。此外, 在火山灰树的树冠上, 不被悬挂在树枝上, DD 的陷阱被放在全太阳下, 5-10 米远离树木繁茂区域边缘的灰树, 或散落在 open-grown 的灰树之中。

<table><tbody><tr><td>Light green corrugated plastic panel: 120 cm x 60 cm</td><td>Great Lakes IPM; www.greatlakesipm.com</td><td>IPM-EAB GR</td><td>All three surfaces of each prism need to be covered with clear insect trapping glue, even if the panels are pre-glued. Pre-glued panels are often not sticky enough to consistently capture or retain EAB beetles.&amp;nbsp; Other clear insect trapping glue products are available but are considerably more difficult to apply.&amp;nbsp;&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Light purple corrugated plastic panel: 120 cm x 60 cm</td><td>Great Lakes IPM; www.greatlakesipm.com</td><td>IPM-EAB&amp;nbsp; LP</td><td></td></tr><tr><td>Large cable tie (4): 60 cm with a 79 kg capacity</td><td>Cabletiesandmore.com; http://www.cabletiesandmore.com/cableties.php</td><td>CT-24-NU-100PK</td><td></td></tr><tr><td>Medium cable ties (4): 20 cm with a 22.7 kg capacity</td><td>Cabletiesandmore.com; http://www.cabletiesandmore.com/cableties.php</td><td>CT261</td><td></td></tr><tr><td>Small cable tie: 10 cm with a 8.2 kg capacity</td><td>Cabletiesandmore.com; http://www.cabletiesandmore.com/cableties.php</td><td>CT204</td><td></td></tr><tr><td>cis-3-hexanol pouch</td><td>Synergy Semiochemicals; http://www.semiochemical.com/html/buprestids.html)</td><td>3136</td><td>Lures used to bait DD traps consist of pouches containing cis-3-hexenol, a non-toxic compound present in ash leaves.&amp;nbsp; One pouch is attached to the lower edge of the top prism using a small cable tie.&amp;nbsp; Each pouch of cis-3-hexenol has a release rate of approximately 50 mg/day. Note that cis-3-hexenol is sometimes written as Z-3-hexenol.&amp;nbsp;</td></tr><tr><td>Aphinity Hexenol</td><td>Sylvar Technologies</td><td></td><td></td></tr><tr><td>Lure GLV4 emerald ash borer</td><td>Chemtica, Heredia, Costa Rica</td><td></td><td></td></tr><tr><td>cis-3-hexanol pouch</td><td>WestGreen Global Technologies; http://www.westgreenglobaltechnologies.com/</td><td></td><td></td></tr><tr><td>Clear insect trapping glue&amp;nbsp;</td><td>Hummert International; http://www.hummert.com/product-details/8196/pestick</td><td>01-3522-1&amp;nbsp;&amp;nbsp;</td><td></td></tr><tr><td>Histoclear II histological clearing agent</td><td>National Diagnostics; www.nationaldiagnostics.com</td><td>HS-202</td><td>Histoclear II will be needed to remove the sticky insect glue from suspect beetles.&amp;nbsp; Other histological clearing agents are available but may not remove the glue and some products dissolve plastic, an important consideration if plastic containers are used for soaking the beetles.&amp;nbsp;</td></tr><tr><td>Histoclear II histological clearing agent</td><td>Great Lakes IPM; www.greatlakesipm.com</td><td>10011</td><td>Histoclear II will be needed to remove the sticky insect glue from suspect beetles.&amp;nbsp; Other histological clearing agents are available but may not remove the glue and some products dissolve plastic, an important consideration if plastic containers are used for soaking the beetles.&amp;nbsp;</td></tr><tr><td>t-post: 1.5 m</td><td>multiple sources</td><td></td><td>A t-post (5 feet tall) (1.5 m) is used to support the PVC pipe.&amp;nbsp;&amp;nbsp;</td></tr><tr><td>post pounder</td><td>multiple sources</td><td></td><td>Use a post pounder to set t-posts into the ground. No additional support is necessary.</td></tr><tr><td>HDPE (high density polyethylene) PVC pipe : 3 m x 10 cm diameter</td><td>multiple sources</td><td></td><td></td></tr><tr><td>Forceps (rigid)</td><td>multiple sources</td><td></td><td>Forceps (tweezers) will be needed to remove suspect beetles from the traps. Rigid forceps work better than flexible forceps.&amp;nbsp;</td></tr><tr><td>Latex gloves</td><td>multiple sources</td><td></td><td>Latex gloves are needed for applying the insect trapping glue to the prisms and for checking the traps to collect EAB beetles.&amp;nbsp;&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Baby oil or baby wipes&amp;nbsp;</td><td>multiple sources</td><td></td><td>Baby oil or baby wipes are helpful for removing the trapping glue from hands and equipment.&amp;nbsp;</td></tr><tr><td>Re-sealable plastic specimen bags: 5 cm x 8 cm&amp;nbsp;</td><td>multiple sources</td><td></td><td>Small re-sealable plastic specimen bags are useful for collecting beetles from traps.&amp;nbsp; Each bag should be labelled, either with pre-made, adhesive labels or with soft felt pens.&amp;nbsp;&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Guides to help with distinguishing EAB from beetles native to North America are available on the national EAB website at www.emeraldashborer.info. &amp;nbsp;</td><td></td><td></td><td></td></tr></tbody></table>

building double-decker traps for early detection of emerald ash borer

翡翠灰钻 (EAB) (吉丁吉丁Fairmaire), 最具破坏性的森林昆虫入侵北美洲, 已杀死数亿森林和景观灰 (水曲柳) 树木。一些人工陷阱设计, 以吸引和捕获 EAB 甲虫已经开发, 以检测, 划定, 并监测虫害。双层 (DD) 陷阱包括两个波纹塑料棱镜, 一个绿色和一个紫色, 连接到一个3米高的聚氯乙烯 (PVC) 管道支持的 t 岗位。在 PVC 管顶部的绿色棱镜是用cis-3-hexenol, 一种由灰叶产生的化合物。两个棱镜的表面都涂有粘性的昆虫胶, 以捕获成年 EAB 甲虫。双层圈闭应该放在灰树附近, 但在开阔的地方, 暴露在阳光下。本文介绍了双层圈闭的构造和放置, 并对田间试验进行了总结, 展示了 DD 陷阱在捕获 EAB 甲虫方面的功效。在最近的一项研究中, EAB 密度相对较低的地方, 双层捕收器比绿色或紫色的棱镜陷阱或绿色漏斗陷阱捕获的 EAB 大得多, 所有这些陷阱都被设计成在火山灰树的树冠上悬挂在树枝上。更大比例的双层陷阱是积极的,即, 捕获至少一个 EAB, 比棱镜陷阱或漏斗陷阱挂在灰树檐篷。

在一个大规模的研究, 三人工陷阱设计以及着灰树部署系统, 10-20 米分开, 横跨一个新出没的16公顷森林地区与非常低密度的 EAB25。人工陷阱设计测试包括紫色棱镜诱饵与麦卢卡油和悬浮和 #62; 3 米从一个分支在灰树的树冠, 3 m 高 dd 陷阱与两个绿色板, 和 DD 陷阱与两个紫色的面板支持的 t 岗位。两种颜色的双层陷阱被诱饵混合的火山灰挥发物, 包括cis...

Watch this Scientific Journal Video about Building Double-decker Traps for Early Detection of Emerald Ash Borer at JoVE.com

Building Double-decker Traps for Early Detection of Emerald Ash Borer

有效的陷阱, 以吸引和捕获翡翠灰蛀虫 (EAB) 是一个关键的因素, 检测和管理这种侵入性害虫。双甲板陷阱, 放置在全太阳附近的灰树, 结合视觉和嗅觉的线索, 更有可能捕捉 EAB 比其他陷阱设计在野外试验。

建筑双层疏水阀早期检测翡翠灰螟

Emerald ash borer (EAB) (Agrilus planipennis Fairmaire), the most destructive forest insect to have invaded North America, has killed hundreds of millions ...

building-double-decker-traps-for-early-detection-of-emerald-ash-borer

Research

JoVE Journal

Environment

7.8K 次观看.  Michigan State University. 有效的陷阱, 以吸引和捕获翡翠灰蛀虫 (EAB) 是一个关键的因素, 检测和管理这种侵入性害虫。双甲板陷阱, 放置在全太阳附近的灰树, 结合视觉和嗅觉的线索, 更有可能捕捉 EAB 比其他陷阱设计在野外试验。

视频: Building Double-decker Traps for Early Detection of Emerald Ash Borer

Technique for Studying Arthropod and Microbial Communities within Tree Tissues

Phloem tissues of pine are habitats for many thousands of organisms. Arthropods and microbes use phloem and cambium tissues to seek mates, lay eggs, rear young, feed, or hide from natural enemies or harsh environmental conditions outside of the tree. Organisms that persist within the phloem habitat are difficult to observe given their location under bark. We provide a technique to preserve intact phloem and prepare it for experimentation with invertebrates and microorganisms. The apparatus is called a &#8216;phloem sandwich&#8217; and allows for the introduction and observation of arthropods, microbes, and other organisms. This technique has resulted in a better understanding of the feeding behaviors, life-history traits, reproduction, development, and interactions of organisms within tree phloem. The strengths of this technique include the use of inexpensive materials, variability in sandwich size, flexibility to re-open the sandwich or introduce multiple organisms through drilled holes, and the preservation and maintenance of phloem integrity. The phloem sandwich is an excellent educational tool for scientific discovery in both K-12 science courses and university research laboratories.

We provide a technique to preserve intact tree phloem and prepare it for observation. We create an apparatus called a phloem sandwich that allows for the introduction and observation of arthropods, microbes, and other organisms that inhabit phloem tissues.

技术树内组织学习节肢动物和微生物群落

Phloem tissues of pine are habitats for many thousands of organisms.  Arthropods and microbes use phloem and cambium tissues to seek mates, lay eggs, rear ...

科研

环境科学

A Technique to Screen American Beech for Resistance to the Beech Scale Insect (Cryptococcus fagisuga Lind.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The disease is initiated by feeding activities of the invasive beech scale insect, Cryptococcus fagisuga, which creates entry points for infection by one of the Neonectria species of fungus.&#160;Without scale infestation, there is little opportunity for fungal infection. Using scale eggs to artificially infest healthy trees in heavily BBD impacted stands demonstrated that these trees were resistant to the scale insect portion of the disease complex1.&#160;Here we present a protocol that we have developed, based on the artificial infestation technique by Houston2, which can be used to screen for scale-resistant trees in the field and in smaller potted seedlings and grafts.&#160;The identification of scale-resistant trees is an important component of management of BBD through tree improvement programs and silvicultural manipulation.

A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind.

Beech bark disease is initiated by feeding activities of the beech scale insect that create fungal entry points in the bark.&#160;Trees that are resistant to the scale insect are also disease resistant.&#160;Here we present the protocol we have developed to screen individual beech trees for beech scale resistance.

一种技术，屏幕美国榉木为抵抗山毛榉介壳虫（<em&gt;隐球菌fagisuga</em&gt;林德。）

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The ...

Soil Sampling and Isolation of Entomopathogenic Nematodes (Steinernematidae, Heterorhabditidae)

Entomopathogenic nematodes (a.k.a. EPN) represent a group of soil-inhabiting nematodes that parasitize a wide range of insects. These nematodes belong to two families: Steinernematidae and Heterorhabditidae. Until now, more than 70 species have been described in the Steinernematidae and there are about 20 species in the Heterorhabditidae. The nematodes have a mutualistic partnership with Enterobacteriaceae bacteria and together they act as a potent insecticidal complex that kills a wide range of insect species.
Herein, we focus on the most common techniques considered for collecting EPN from soil. The second part of this presentation focuses on the insect-baiting technique, a widely used approach for the isolation of EPN from soil samples, and the modified White trap technique which is used for the recovery of these nematodes from infected insects. These methods and techniques are key steps for the successful establishment of EPN cultures in the laboratory and also form the basis for other bioassays that consider these nematodes as model organisms for research in other biological disciplines. The techniques shown in this presentation correspond to those performed and/or designed by members of S. P. Stock laboratory as well as those described by various authors.

Soil Sampling and Isolation of Entomopathogenic Nematodes Steinernematidae, Heterorhabditidae

Entomopathogenic nematodes are soil-inhabiting roundworms that parasitize a wide range of insects. We demonstrate sampling methods used for the isolation of these nematodes from the soil using two techniques: the insect baiting and the modified White trap, for the recovery of nematodes from soil samples and infected insect cadavers, respectively.

土壤采样及昆虫病原线虫的分离（Steine​​rnematidae，异小）

Entomopathogenic nematodes (a.k.a. EPN) represent a group of soil-inhabiting nematodes that parasitize a wide range of insects. These nematodes belong to ...

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.

Preventing colonization of bird nest boxes by invasive Africanized honey bees is important for conservation efforts of nest-site limited birds. We provide an integrated pest management approach to &#34;push&#34; bees away from nest boxes with a repellant insecticide, permethrin, and &#34;pull&#34; them toward pheromone baited swarm traps.

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. ...

The Hawaii Protocol for Scientific Monitoring of Coffee Berry Borer: a Model for Coffee Agroecosystems Worldwide

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at capturing and quantifying the dynamics and impact of this invasive insect pest as well as the development of its host plant across a heterogeneous landscape. The cornerstone of this comprehensive monitoring system is timely georeferenced data collection on CBB movement, coffee berry infestation, mortality by the fungus Beauveria bassiana, and coffee plant phenology via a mobile electronic data recording application. This electronic data collection system allows field records to be georeferenced through built-in global positioning systems, and is backed by a network of weather stations and records of farm management practices. Comprehensive monitoring of CBB and host plant dynamics is an essential part of an area-wide project in Hawaii to aggregate landscape-level data for research to improve management practices. Coffee agroecosystems in other parts of the world that experience highly variable environmental and socioeconomic factors will also benefit from implementing this protocol, in that it will drive the development of customized integrated pest management (IPM) to manage CBB populations.

Comprehensive monitoring of coffee berry borer and host plant dynamics is essential for aggregating landscape-level data to improve management of this invasive pest. Here, we present a protocol for scientific monitoring of coffee berry borer movement, infestation, mortality, coffee plant phenology, weather, and farm management via a mobile electronic data recording application.

夏威夷咖啡莓科学监测协议: 全球咖啡农田模型

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at ...

A Precise and Autonomous System for the Detection of Insect Emergence Patterns

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of emerging insects they can detect. In order to obtain precise measurement of insect emergence, it is necessary for systems to be semi-automated and able to measure large numbers of emerging insects. We addressed these issues by designing and building a system that is automated and can measure emergence of up to 1200 insects. We modified the existing &#34;falling-ball&#34; system using Arduino microcontrollers to automate data collection and expand the sample size through multiple data channels. Multiple data channels enable the user to not only increase their sample size, but also allows for multiple treatments to be run simultaneously in a single experiment. Furthermore, we created an R script to automatically visualize the data as a bubble plot, while also calculating the median day and time of emergence. The current system was designed using 3D printing so the user can modify the system to be adjusted for different species of insects. The goal of this protocol is to investigate important questions in chronobiology and stress physiology, using this precise and automated system to measure insect emergence patterns.

Measurement of insect emergence patterns requires precision. Existing systems are only semi-automated and sample size is limited. We addressed these issues by designing a system using microcontrollers to precisely measure the time of emergence of large numbers of emerging insects.

一种精确、自主的昆虫产生模式检测系统

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of ...

A Method for Quantifying Foliage-Dwelling Arthropods

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density requires a method with high detection probability and a known sampling area. While most described methods provide a qualitative or semi-quantitative estimate adequate for describing species presence, richness, and diversity, few provide an adequately consistent detection probability and known or consistent sampling areas to provide an index or estimate with adequate precision to detect differences in abundance across environmental, spatial, or temporal variables. We describe how to quantify leaf-dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate and quantify them. As we demonstrate, this method can be used at a landscape scale to quantify leaf-dwelling arthropods with adequate precision to test for and describe how spatial, temporal, environmental, and ecological variables influence arthropod richness and abundance. This method allowed us to detect differences in density, richness, and diversity of leaf-dwelling arthropods among 5 genera of trees commonly found in southeastern deciduous forests.

We describe how to quantify leaf dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate for quantification.

定量叶类节肢动物的方法

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density ...

Quantifying Corticolous Arthropods Using Sticky Traps

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimate of density requires a method with high detection probability and a consistent sampling area. We used manufactured sticky traps to compare abundance, total length (a surrogate for biomass), richness, and Shannon diversity of corticolous arthropods among the boles of 5 tree species. Efficacy of this method was adequate to detect variation in corticolous arthropods among tree species and provide a standard error of the mean that was &#60;20% of the mean for all estimates with sample sizes from 7 to 15 individual trees of each species. Our results indicate, even with these moderate sample sizes, the level of precision of arthropod community metrics produced with this approach is adequate to address most ecological questions regarding temporal and spatial variation in corticolous arthropods. Results from this method differ from other quantitative approaches such as chemical knockdown, visual inspection, and funnel traps in that they provide an indication of corticolous arthropod activity over a relatively long-term, better including temporary bole residents, flying arthropods that temporarily land on the tree bole and crawling arthropods that use the tree bole as a travel route from the ground to higher forest foliage. Furthermore, we believe that commercially manufactured sticky traps provide more precise estimates and are logistically simpler than the previously described method of directly applying a sticky material to tree bark or applying a sticky material to tape or other type of backing and applying that to the tree bark.

We describe a semi-quantitative approach of measuring characteristics of corticolous (bark-dwelling) arthropod communities. We placed commercially manufactured sticky traps on tree boles to estimate abundance, total length (a surrogate to biomass), richness, and Shannon diversity for comparison among tree species.

使用粘性陷阱量化皮质节肢动物

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimate of density ...

Low-Cost Automated Flight Intercept Trap for the Temporal Sub-Sampling of Flying Insects Attracted to Artificial Light at Night

Sampling methods are selected depending on the targeted species or the spatial and temporal requirements of the study. However, most methods for passive sampling of flying insects have a poor temporal resolution because it is time-consuming, costly and/or logistically difficult to perform. Effective sampling of flying insects attracted to artificial light at night (ALAN) requires sampling at user-defined time points (nighttime only) across well-replicated sites resulting in major time and labor-intensive survey effort or expensive automated technologies. Described here is a low-cost automated intercept trap that requires no specialist equipment or skills to construct and operate, making it a viable option for studies that require temporal sub-sampling across multiple sites. The trap can be used to address a wide range of other ecological questions that require a greater temporal and spatial scale than is feasible with previous trap technology.

To study the impacts of artificial light at night (ALAN) on nocturnal flying insects, sampling needs to be confined to nighttime. The protocol describes a low-cost automated flight intercept trap that allows researchers to sample at user-defined periods with increased replication.

一种低成本自动飞行拦截陷阱，用于夜间被人造光吸引的飞虫进行时间子采样

Sampling methods are selected depending on the targeted species or the spatial and temporal requirements of the study. However, most methods for passive ...

Analysis of Gene Expression in Emerald Ash Borer (Agrilus planipennis) Using Quantitative Real Time-PCR

Emerald ash borer (EAB, Agrilus planipennis) is an exotic invasive pest, which has killed millions of ash trees (Fraxinus spp) in North America. EAB continues to spread rapidly and attacks ash trees of different ages, from saplings to mature trees. However, to date very little or no molecular knowledge exists for EAB. We are interested in deciphering the molecular-based physiological processes at the tissue level that aid EAB in successful colonization of ash trees. In this report we show the effective use of quantitative real-time PCR (qRT-PCR) to ascertain mRNA levels in different larval tissues (including midgut, fat bodies and cuticle) and different developmental stages (including 1st-, 2nd-, 3rd-, 4th-instars, prepupae and adults) of EAB. As an example, a peritrophin gene (herein named, AP-PERI1) is exemplified as the gene of interest and a ribosomal protein (AP-RP1) as the internal control. Peritrophins are important components of the peritrophic membrane/matrix (PM), which is the lining of the insect gut. The PM has diverse functions including digestion and mechanical protection to the midgut epithelium.

Analysis of Gene Expression in Emerald Ash Borer Agrilus planipennis Using Quantitative Real Time-PCR

Quantitative real-time PCR (qRT-PCR) is an effective tool to diagnose mRNA levels in different insect tissues and developmental stages. In this report we show the use of qRT-PCR to ascertain mRNA levels in different larval tissues and developmental stages of the invasive insect species, emerald ash borer.

白蜡窄吉丁的基因表达分析（ Agrilus planipennis）使用实时定量PCR

Emerald ash borer (EAB, Agrilus planipennis) is an exotic invasive pest, which has killed millions of ash trees (Fraxinus spp) in North America. EAB ...

建筑双层疏水阀早期检测翡翠灰螟

摘要

探索更多视频

此视频中的章节

技术树内组织学习节肢动物和微生物群落

一种技术，屏幕美国榉木为抵抗山毛榉介壳虫（

土壤采样及昆虫病原线虫的分离（Steinernematidae，异小）

推挽议定书蜜蜂会影响家禽的殖民化巢箱

夏威夷咖啡莓科学监测协议: 全球咖啡农田模型

一种精确、自主的昆虫产生模式检测系统

定量叶类节肢动物的方法

使用粘性陷阱量化皮质节肢动物

一种低成本自动飞行拦截陷阱，用于夜间被人造光吸引的飞虫进行时间子采样

白蜡窄吉丁的基因表达分析（ Agrilus planipennis）使用实时定量PCR

建筑双层疏水阀早期检测翡翠灰螟

摘要

探索更多视频

此视频中的章节

技术树内组织学习节肢动物和微生物群落

一种技术，屏幕美国榉木为抵抗山毛榉介壳虫（

土壤采样及昆虫病原线虫的分离（Steine​​rnematidae，异小）

推挽议定书蜜蜂会影响家禽的殖民化巢箱

夏威夷咖啡莓科学监测协议: 全球咖啡农田模型

一种精确、自主的昆虫产生模式检测系统

定量叶类节肢动物的方法

使用粘性陷阱量化皮质节肢动物

一种低成本自动飞行拦截陷阱，用于夜间被人造光吸引的飞虫进行时间子采样

白蜡窄吉丁的基因表达分析（ Agrilus planipennis）使用实时定量PCR

土壤采样及昆虫病原线虫的分离（Steinernematidae，异小）