从个体到群体度评价环境化学物质对蜜蜂发展的影响
Summary
在这里我们提出 的方法，以农药污染的食物输送到两个单独的蜜蜂和蜂箱菌落。过程评估由基本幼虫饮食的体内喂养和也对蜂窝菌落的天然条件对个体蜜蜂的农药效果。
Abstract
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.
Introduction
农药在环境中的存在是最严重的问题之一是影响一个蜜蜂1，2，3的使用寿命。一些研究表明农药残留量的蜂蜜蜂群和蜜蜂产品的共同存在。在台湾，农药的平均应用为11-12公斤/公顷，每年（2005至13年）。农药在台湾使用量是比欧盟国家的高，以及拉丁美洲国家的4，5。换句话说，在养蜂环境正在遭受严重的农药压力，特别是在台湾，可能在其他国家。
在西方蜜蜂是农业系统6的主要传粉昆虫之一，它也产生有价值的产品，如蜂蜜。然而，蜜蜂是博览会ED各种杀虫剂和这些农药能觅食上采集花蜜和花粉7,8时已喷洒杀虫剂花后带回蜂箱。他们还可以通过自己的目的来控制荨麻疹9，10，11内的虫害问题养蜂人暴露于杀虫剂。由于蜜蜂幼虫被护士蜜蜂为他们的发展，幼虫，无人机喂，甚至女王可能会接触到这些农药污染的花蜜和花粉12。需要解决的13多种农药对蜜蜂的毒性。
许多已作出努力，以评估环境农药残留的问题。杨等人。测试的蜜蜂幼虫在发展中的神经毒性杀虫剂吡虫啉的影响蜂箱和报道，吡虫啉亚致死剂量导致成年蜂14的嗅觉关联行为。此外，Urlacher 等。检查的有机磷农药，毒死蜱，对蜜蜂工人的学习表现在实验室条件下15的亚致死效应。在我们以前的研究中，我们评估的昆虫生长调节剂（IGR）的影响，吡丙醚（PPN），幼虫蜜蜂16。
在本文中，我们提出，以评估对蜜蜂的发展化学影响的方法。进行了描述并应用于任何个别蜜蜂或菌落蜜蜂馈送方法。起初，我们在殖民地幼虫上测试了不同浓度的农药污染的基本幼虫饲料（BLD），以评估在体内个别蜜蜂农药的影响。我们接着来模拟自然康迪特通过使用蜂箱内农药污染的糖浆中的农药的离子。在该方法中，PPN，它被广泛用于对抗有害昆虫17和有害蜜蜂幼虫和蛹16，18，19的发展，将表示该领域的农药的负面影响的指标。
Protocol
Representative Results
Discussion
王后限制产蛋方法和王后交换法是用于获得本协议内的现场测试设置蜜蜂基团的关键步骤。王后限制产蛋方法允许蜜蜂的生命周期的同步。因此，研究人员可以选择相同的年龄与不同剂量农药的治疗1日龄幼虫。对于女王交换法，王后被部分A（4帧）和B（5帧）以获得蜜蜂的不同发育阶段的现场测试，以评估农药和农药残留的影响之间进行交换。此外，大量的选择育雏细胞被记录在使用透明幻灯片?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项研究是由格兰特105AS-13.2.3-BQ-B1从动物局动植物卫生检验检疫，农业委员会，行政院和格兰特103-2313-B-197-002-MY3从支持部科学技术部（MOST）。
Materials
| Honey bee box | SAN-YI Honey Factory | W1266 | Honeybees rearing |
| Queen excluder (between frames) | SAN-YI Honey Factory | I1575 | Queen limitation |
| Queen excluder (on top ) | SAN-YI Honey Factory | I1566 | Queen limitation on top |
| Bee brush | SAN-YI Honey Factory, Taiwan | W1414 | clean the bees on frame gently |
| Bee feeder | SAN-YI Honey Factory, Taiwan | P0219 | feed sugar syrup to colony |
| Transparent slide | Wan-Shih-Chei, Taiwan (http://www.mbsc.com.tw/a01goods.asp?s_id=40) | 1139 | Mark the larval area on the frames (Material: Polyethylene Terephthalate, PET) (Size= Length*Width*thick= 29.7mm*21mm*0.1mm) |
| 24 well tissu culture plate | Guangzhou Jet Bio-Filtration Co., Ltd | TCP011024 | Rearing pupae from extraction |
| Autoclave | Tomin medical equipmenco., LTD. | TM-321 | Make sterilized distilled deionized water (ddH2O) |
| P20 pipetman | Gilson | F123600 | Add PPN into bee larval food pool |
| Incubator | Yihder Co., Ltd. | LE-550RD | Rearing pupae from extraction |
| Kimwipes | COW LUNG INSTRUMENT CO., LTD | KCS34155 | Rearing pupae from extraction |
| Royal jelly | National Ilan University (NIU) | NIU | Make basic larval diet (BLD) |
| D-(+)-Glucose | Sigma | G8270 | Make basic larval diet (BLD) |
| D-(-)-Fructose | Sigma | F0127 | Make basic larval diet (BLD) |
| Yeast extract | CONDA, pronadisa | 1702 | Make basic larval diet (BLD) |
| Sucrose | Taiwan sugar coporation | E01071010 | Make sugar syrup for bee food |
| Pyriproxyfen (11%) | LIH-NUNG CHEMICAL CO.. LTD. | Registration No. 1937 | Insect growth regulator (IGR) used in the experiment |
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