Name: a 3d printed pollen trap for bumble bee (bombus) hive entrances
Uploaded: 2020-07-09T14:30:00.000+00:00
Duration: 7 min 19 s
Description: Watch this Scientific Journal Video about A 3D Printed Pollen Trap for Bumble Bee Bombus Hive Entrances at JoVE.com

我们感谢Colby Carpenter和Spencer Mathias在3D打印设计方面的帮助。我们感谢艾伦·克林格（Ellen Klinger）协助制作摄影人物，并感谢乔纳森·B·科赫（Jonathan B. Koch）协助进行修订。资金由美国农业部 - ARS授粉昆虫生物学，管理和系统学研究部门提供。

1. 打印花粉陷阱结构
<ol><li>下载大黄蜂筑巢的巢箱的相应STL文件（例如，Biobest或Koppert风格的蜂巢，https://www.ars.usda.gov/pacific-west-area/logan-ut/pollinating-insect-biology-management-systematics-research/docs/pollen-traps/）。这些文件向公众开放，最终用户可以免费下载和修改。</li>
	<li>在打印机程序中打开 STL 文件。按照打印机制造商的说明构建四个陷印组件。 注:打印疏水阀体约 3 小时，打印集水盆约 2 小时，滤镜和陷印盒闭合插入物各 30 分钟。疏水阀主体尺寸为 6 厘米 x 3.8 厘米 x 7 厘米。</li>
</ol>2. 花粉捕集器组件
<ol><li>取下印有疏水阀体和集水池的支撑结构，包括疏水阀体的筛分结构（图1）。</li>
	<li>使用安装在手钻中的 3/16 英寸（0.476 厘米）钻头清除穿过花粉过滤器凸起边缘的任何塑料股，这些塑料股可能会阻碍蜜蜂通过过滤器孔。使用箱式切割剃须刀片和砂纸，将花粉过滤器平坦侧的任何凸起或凸起的边缘均匀。</li>
	<li>将花粉过滤器轻轻推过陷阱主体的一侧，将花粉过滤器放入疏水阀体中。过滤器只能以一种方式安装，因为疏水阀的左侧有一个更大的开口，以适应凸起的过滤器锥体的通过。<ol>
<li>如果侧缝太小，花粉过滤器无法顺利滑过，请使用剃须刀或其他可以一次去除一小部分塑料的工具，从疏水阀体的狭缝中刮出足够的塑料。确保花粉过滤器牢固地安装到位，花粉过滤器和捕集器体之间的间隙不超过2 mm。</li>
	</ol>
</li>
	<li>通过将捕集器主体底部的凸起边缘滑入集水盆顶部的凹槽中，将集水盆连接到捕集器主体上。集水池应直接位于疏水阀体的筛分区域下方（图 1A\u2012E）。</li>
	<li>通过切割或打磨塑料进行适当的修改，以便从疏水阀体上平滑放置和移除集水盆。集水池的放置会将花粉过滤器固定到位，在收集盆被移除之前，花粉过滤器无法移除，在集水盆就位时也无法插入花粉过滤器。 注意:如果将多个蜂箱彼此靠近放置，除了部署具有不同方向的蜂巢外，为每个蜂巢提供陷阱体，集水盆和花粉过滤器结构的独特颜色组合将有助于返回的工人找到巢穴。</li>
</ol>3. 大黄蜂蜂群准备
<ol><li>在部署菌落之前，停止花粉喂养24\u201248小时。这将导致工蚁用尽任何储存的花粉，并刺激它们离开巢穴寻找花粉。</li>
	<li>通过将疏水阀闭合插件插入过滤器槽中，防止蜜蜂在安装疏水阀时逃逸，准备用于安装蜂巢陷阱体。</li>
	<li>在红灯下工作以防止蜜蜂飞翔，将塑料巢箱从纸板外箱中取出。</li>
	<li>找到巢前方的塑料巢入口。根据巢供应商的不同，有两种入口样式:Koppert样式的盒子（步骤3.5）和Biobest风格的盒子（步骤3.6）。</li>
	<li>对于Koppert风格的蜂巢入口，通过拉动入口标签直到两个入口孔都打开，将花粉陷阱安装到巢穴入口处。<ol>
<li>将花粉捕集器的两根管子插入入口孔中，确保花粉捕集器的筛子在底部。轻轻向下推塑料入口卡舌，将花粉陷阱固定到位。</li>
	</ol>
</li>
	<li>要将花粉陷阱安装到Biobest式蜂巢入口处，请使用平头螺丝刀从巢箱中轻轻撬出塑料入口装置。将花粉陷阱插入巢穴入口孔中，直到花粉陷阱牢固地贴在巢穴上（图1E）。<ol>
<li>如果需要，使用胶带或速干胶将疏水阀固定在巢穴中，陷阱与巢箱接触。</li>
	</ol>
</li>
	<li>将塑料巢箱放回纸板箱。纸板可能需要切掉以容纳花粉陷阱。</li>
</ol>4. 巢穴的部署
<ol><li>将巢箱放入研究区域。提供覆盖物以防止降水和风锚定，因为这些会对收集的花粉的质量和数量产生不利影响。为放置在温室中的蜂箱提供足够的阳光覆盖，以减少过热。</li>
	<li>从陷阱体上取下陷阱封闭插件，让蜜蜂自由觅食，以便觅食者可以根据周围区域和巢穴的位置定位自己。在正常情况下，定向飞行时间应在24小时内完成。</li>
</ol>5. 花粉采集
<ol><li>要接合疏水阀，请将花粉过滤器滑入过滤器槽，确保其牢固到位。</li>
	<li>通过从正面将集水盆滑到疏水阀体上直到其完全关闭来安装集水盆。如果集水盆过度松动或从疏水阀体上脱落，请使用橡皮筋将其固定在疏水阀体上。</li>
	<li>观察蜜蜂在首次部署时进入和退出花粉陷阱，以确保花粉过滤器孔足够大以容纳蜜蜂。<ol>
<li>如果工人无法通过花粉过滤器，请取下过滤器，然后使用大于3/16英寸（0.476厘米）的钻头来增加孔的尺寸。按顺序进行，每次增加孔径1/32英寸（0.079厘米），因为太大的孔不会收集任何花粉。</li>
		<li>一旦蜜蜂能够通过过滤器，继续观察入口，以确保花粉在重新进入时被去除。 注意:蜜蜂在花粉过滤器孔中移动应该有些困难，尤其是它们通过的头几次。如果它们太容易通过，花粉可能不会从corbiculae中脱落。</li>
	</ol>
</li>
	<li>在指定的花粉收集期后，从捕集器体中滑动并取出集水盆。</li>
	<li>根据您的实验设计处理花粉负载。</li>
	<li>取下花粉过滤器，让工人自由觅食，直到下一个花粉采集期。在实验期间，陷阱体可能仍然附着在蜂巢上。 注意:只要研究人员希望从蜂群中收集花粉，花粉陷阱就可以参与。然而，在一周内部署花粉诱捕器超过24小时可能导致蜂巢中的育雏饥饿并延缓菌落发育。</li>
</ol>

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作者没有什么可透露的。

从大黄蜂群入口收集花粉可以进行各种生态和农业研究。确定大黄蜂收集花粉的花卉来源为有助于殖民地整体饮食的植物多样性提供了有价值的信息和见解19。确定花粉来源对农业生产和野生土地生态系统服务的研究都有影响12，20。通过收集相对较大的样本大小的皮质花粉负载，研究人员可以确定大黄蜂是否在它们部署的目标作物<sup ...

大黄蜂（Bombus spp.）是大型健壮的昆虫，遍布世界温带，高山和北极地区1。它们对植物群落很重要，并为它们访问的农作物提供重要的授粉服务2.最近几个物种的丰度和分布的下降使它们作为传粉媒介的重要性成为公众意识的前沿3.研究人员已经确定了几个可能导致人口下降的压力因素，包括缺乏多样和丰富的花卉资源，大黄蜂以此为食4。确定大黄蜂从哪种植物物种中觅食，使研究人员和土地管理人员能够了解大黄蜂如何应对资源可用性，竞争和人为干扰的变化5，6。
调查大黄蜂花粉觅食偏好的研究通常由研究人员进行，研究人员捕捉在花朵处觅食的个体蜜蜂，然后从标本中去除皮质花粉负荷以进行进一步处理和鉴定7，8，9，10。虽然这种方法可以深入了解一个物种或大黄蜂物种的组合如何利用区域7中的资源，但它是时间密集型的，如果没有额外的分子分析来确定觅食蜂11的起源殖民地，就无法辨别蜂巢之间偏好的潜在差异。
对于一些觅食动力学的研究，希望在单个殖民地进行研究;然而，野生大黄蜂巢通常位于地下或地面，因此很难找到12。商业生产的大黄蜂蜂巢为研究人员提供了更多的访问和更好的实验控制，并且从工人身上去除花粉仍然主要通过捕获觅食者返回蜂巢并手动去除其Corbicular花粉负载13，14。从蜜蜂的鞘翅中手工去除花粉是时间密集型的，每小时的花粉产量很低，特别是在蜂巢入口处，返回花粉觅食者的比率可能很低。此外，手动清除蜜蜂的花粉会导致受到干扰的工人被蜇伤。
花粉陷阱已被用于实验性地去除蜜蜂的花粉数十年15;然而，一种从大黄蜂中去除花粉的被动方法尚未开发出来。开发一种机制以从返回的觅食者大黄蜂中去除花粉的主要障碍是大黄蜂群中存在的工蜂大小的巨大变化16。蜜蜂花粉陷阱之所以有效，很大程度上是因为蜜蜂工人的大小变化不大。此外，这些陷阱在安装后只需要轻微的操作，并且不需要牺牲蜜蜂17。这是使用屏幕或塑料表面实现的，当工人返回蜂巢时，这些屏幕或塑料表面会将花粉从工人的后腿上移开。这些诱捕器仅从返回的觅食者身上去除一部分花粉负荷，并且这些诱捕器的各种设计导致花粉收集效率不同。当花粉从蜂腿上取出时，它通过屏幕落入蜜蜂无法进入的收集盆中，因此研究人员可以在对蜂巢产生轻微干扰的情况下将其移除。
本研究的目的是调整用于从蜜蜂蜂巢中收集花粉的技术，并使用3D打印结构将其应用于大黄蜂巢，并在 Bombus huntii的菌落上测试陷阱设计。设计过程遵循的假设是，陷阱的生产成本应该低廉，适应各种大黄蜂物种，对蜜蜂造成的伤害或干扰最小，花粉去除率应超过手工收集的花粉。三维打印技术用途广泛，易于访问，并且是一种具有成本效益的工具，允许研究人员为特定目的复制和修改对象18。这里介绍的技术指示用户建立花粉陷阱，并将它们附着在市售的大黄蜂群落上。这些陷阱不是为野生殖民地设计的。这些陷阱被动地从携带大黄蜂的花粉的后腿上去除皮质花粉负荷，因为它们返回巢箱。

<table><tbody><tr><td>MakerBot Replicator+</td><td>MakerBot</td><td>Model PABH65</td><td></td></tr><tr><td>MakerBot Tough Material</td><td>PLA Filament</td><td></td><td>various colors</td></tr><tr><td>Nest Box</td><td>Biobest</td><td></td><td>Not sold publicly without bee purchase</td></tr></tbody></table>

a 3d printed pollen trap for bumble bee (bombus) hive entrances

为了验证大黄蜂觅食花粉的植物来源，必须收集个体以去除其皮质花粉负荷以进行分析。 传统上，这是通过在巢穴入口处或花朵上网捕觅食者来完成的，在冰上冷却蜜蜂，然后用镊子或刷子从corbiculae中去除花粉负载。 这种方法是时间和劳动密集型的，可能会改变正常的觅食行为，并可能导致执行任务的工人发生刺痛事件。 花粉陷阱，例如用于蜜蜂蜂箱的花粉陷阱，通过在通过巢穴入口处的屏幕时将工人腿部的皮质花粉负载移开来收集花粉。 陷阱可以用最少的劳动力从返回的觅食蜂中去除大量花粉，但迄今为止还没有这样的陷阱可用于大黄蜂群落。大黄蜂蜂群内的工蚁大小可能不同，使得入口的大小选择难以使这种机制适应商业饲养的大黄蜂蜂巢。 使用3D打印设计程序，我们创建了一个花粉陷阱，成功地从返回的大黄蜂觅食者的腿上移除了皮质花粉负荷。 这种方法大大减少了研究人员从返回蜂群的大黄蜂觅食者那里收集花粉所需的时间。我们介绍了设计，花粉去除效率测试的结果，并为调查人员建议了修改区域，以使陷阱适应各种大黄蜂物种或巢箱设计。

测试了八种不同的花粉过滤器设计，以确定它们在从返回的大黄蜂工人中去除皮质花粉负荷方面的功效和效率。所有设计都成功地从返回的觅食者身上消除了至少一个皮质花粉负荷。然而，有些被发现会减慢工人离开或进入蜂巢的速度，或者无法去除花粉负荷（表1）。在温室中生长的Phacelia tanacetifolia上的4个实验室饲养的B. huntii Greene菌落上依次测试了带有各种过滤器的?...

Watch this Scientific Journal Video about A 3D Printed Pollen Trap for Bumble Bee Bombus Hive Entrances at JoVE.com

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

我们提出了一种非致命和自动化的机制，从返回蜂巢的大黄蜂（Bombus）工人那里收集花粉。包括有关生产、准备、安装和使用设备的说明。通过使用3D打印对象，对设计的修改是及时，有效的，并允许快速周转进行测试。

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.  ...

a-3d-printed-pollen-trap-for-bumble-bee-bombus-hive-entrances

Research

JoVE Journal

Biology

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

5.1K 次观看.  Utah State University. 我们提出了一种非致命和自动化的机制，从返回蜂巢的大黄蜂（Bombus）工人那里收集花粉。包括有关生产、准备、安装和使用设备的说明。通过使用3D打印对象，对设计的修改是及时，有效的，并允许快速周转进行测试。

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

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. ...

科研

环境科学

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 ...

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 ...

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 ...

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 ...

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees (Apis mellifera L.)

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker bees spend the first half of their life inside the dark hive, where they are nursing the larvae or building the regular hexagonal combs for food (e.g. pollen or nectar) and brood3. The antennae are extraordinary multisensory feelers and play a pivotal role in various tactile mediated tasks4, including hive building5 and pattern recognition6. Later in life, each single bee leaves the hive to forage for food. Then a bee has to learn to discriminate profitable food sources, memorize their location, and communicate it to its nest mates7. Bees use different floral signals like colors or odors7, 8, but also tactile cues from the petal surface9 to form multisensory memories of the food source. Under laboratory conditions, bees can be trained in an appetitive learning paradigm to discriminate tactile object features, such as edges or grooves with their antennae10, 11, 12, 13. This learning paradigm is closely related to the classical olfactory conditioning of the proboscis extension response (PER) in harnessed bees14. The advantage of the tactile learning paradigm in the laboratory is the possibility of combining behavioral experiments on learning with various physiological measurements, including the analysis of the antennal movement pattern.

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees Apis mellifera L.

In this protocol we show how to condition harnessed honey bees to tactile stimuli and introduce a 2D motion capture technique for analyzing the kinematics of fine-scale antennal sampling pattern.

触觉空调和取样策略在蜜蜂触角的运动分析（<em&gt;蜜蜂</em&gt; L.）

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker ...

神经科学

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). ...

Radio Frequency Identification and Motion-sensitive Video Efficiently Automate Recording of Unrewarded Choice Behavior by Bumblebees

We present two methods for observing bumblebee choice behavior in an enclosed testing space. The first method consists of Radio Frequency Identification (RFID) readers built into artificial flowers that display various visual cues, and RFID tags (i.e., passive transponders) glued to the thorax of bumblebee workers. The novelty in our implementation is that RFID readers are built directly into artificial flowers that are capable of displaying several distinct visual properties such as color, pattern type, spatial frequency (i.e., &#8220;busyness&#8221; of the pattern), and symmetry (spatial frequency and symmetry were not manipulated in this experiment). Additionally, these visual displays in conjunction with the automated systems are capable of recording unrewarded and untrained choice behavior. The second method consists of recording choice behavior at artificial flowers using motion-sensitive high-definition camcorders. Bumblebees have number tags glued to their thoraces for unique identification. The advantage in this implementation over RFID is that in addition to observing landing behavior, alternate measures of preference such as hovering and antennation may also be observed. Both automation methods increase experimental control, and internal validity by allowing larger scale studies that take into account individual differences. External validity is also improved because bees can freely enter and exit the testing environment without constraints such as the availability of a research assistant on-site. Compared to human observation in real time, the automated methods are more cost-effective and possibly less error-prone.

This video describes Radio-Frequency Identification (RFID) and motion-sensitive video recording methods to monitor choice behavior by bumblebees.

无线射频识别和动作感应视频高效地自动Unrewarded选择行为的记录由大黄蜂

We present two methods for observing bumblebee choice behavior in an enclosed testing space. The first method consists of Radio Frequency Identification ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

一种新的行为分析调查个人的味觉，可自由移动的熊蜂（<em&gt;欧洲熊蜂</em&gt;）

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

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 ...

用于大黄蜂（Bombus）蜂巢入口的3D打印花粉陷阱

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