我们实验室的研究目前由澳大利亚研究理事会 (ARC、DP170100008 和 DP180100144)、美国空军科学研究办公室 (AFOSR、FA9550-15-1-0188) 和 Stiftelsen Olle Engkvist (2016/348) 资助。我们感谢过去的实验室成员为开发食蚜虻库存, Cederholms Lantbruk 和 & T 绿色 & 儿子提供了在他们的农场上的牛粪和 hoverflies, 以及阿德莱德和乌普萨拉植物园的收集许可证和持续的支持。

1. 建立俘虏的 E. 管殖民地
<ol><li>通过收集成熟的幼虫 (步骤 1.2) 或通过收集自由飞行 hoverflies (步骤 1.3) 建立殖民地。</li>
	<li>
成熟幼虫的收集
	<ol>
<li>从牛场的粪便中收集第二和第三龄幼虫。 注意: 成熟的幼虫在迁徙阶段开始时最容易发现, 因为他们正积极寻求一个黑暗的干燥环境来蛹。这往往是在肥料坑的边界附近, 潮湿的肥料接近干燥地区含有大量的秸秆。我们在瑞典乌普萨拉附近的一个畜牧场得到许可。</li>
		<li>在步骤3.2 中概述的奶牛粪便后成熟幼虫。</li>
	</ol>
</li>
	<li>
野生 Hoverflies 的收集
	<ol>
<li>收集野外 hoverflies, 通常来自植物园和公园, 那里有大量的开花植物。 注: 我们得到了几个地点的许可, 包括三阿德莱德植物园, Myponga, 南澳大利亚的一个奶牛场, 以及在瑞典乌普萨拉各地的各种植物园和公园。</li>
		<li>房屋领域收集 hoverflies 如步骤2所述。</li>
	</ol>
</li>
</ol>2. Hoverflies 的住房和长期维护
<ol><li>房子 hoverflies 在 30 cm x 45 cm 塑料袋子为小组20或更少, 或者在昆虫饲养笼子 (25 cm x 25 cm x 25 cm) 为更大的小组。</li>
	<li>提供食物和水的ad 随意, 形式为10-20 粒蜂花粉和2-3 毫升蜂蜜放置在几个湿润的棉花球顶部。 注: 对于塑料袋中的外壳, 重要的是, 棉花球是湿润的, 但不是过度饱和, 因为任何堆积的水袋内可能会损害生存率。相反, 在饲养昆虫的笼子里, 网边允许大量的蒸发发生。因此, 棉球应放置在浅容器中, 并充分饱和。<ol>
<li>让 hoverflies 在室温下进食6小时。</li>
		<li>安置 hoverflies 与食物和水在他们的住房, 在冰箱在8-10 °c 和保持在完全黑暗中。 注意: 在黑暗中储存 hoverflies 8-10 摄氏度会导致 hoverflies 进入冬眠状态, 活动和新陈代谢率降低。</li>
	</ol>
</li>
	<li>每3-4 天从冰箱中取出 hoverflies, 从而打破人工冬眠, 并允许喂养和梳理发生。<ol>
<li>将 hoverflies 转移到新的塑料袋或干净的昆虫笼中, 新鲜的食物和水。这种转移可以手动完成, 对于小数目的苍蝇, 或利用趋光性, 为较大的数字。</li>
		<li>要利用趋光性, 把干净的昆虫笼加入旧的笼子里, 确保 hoverflies 在两者之间自由移动, 而不逃避。用不透明的布料盖住旧的昆虫笼。hoverflies 将走向光明, 进入清洁的昆虫笼。</li>
	</ol>
</li>
	<li>允许 hoverflies 在室温下喂饲和新郎6小时。</li>
	<li>返回 hoverflies, 与食物和水在他们的住房, 到冰箱在8-10 °c 在完全黑暗中。这 recommences 了 hoverflies 的人工冬眠。</li>
	<li>继续周期性打破冬眠, 每3-4 天, 以确保在他们圈养期间 hoverflies 的健康和长寿。</li>
</ol>3. 对管的实验室饲养
<ol><li>从牛场收集的后成熟幼虫 (步骤 3.2) 或由妊娠野生捕获雌性 (步骤 3.3) 放置的后卵。</li>
	<li>
奶牛场成熟幼虫的实验室饲养
	<ol>
<li>将幼虫从奶牛场中收集到30升桶中的牛粪中。</li>
		<li>放置桶, 包含成熟的幼虫, 在一个更大的盒子或袋子 (最小容积50-60 升) 和地方20-30 升木屑的高度的边缘的桶。 注意: 这允许 3rd龄幼虫爬入木屑和蛹。</li>
		<li>在天花板上挂一个双层蚊帐, 允许它在盒子和/或袋子上悬垂, 从而确保幼虫和任何新兴的 hoverflies 都不能逃脱。 注意: 作为预防措施, 双面胶带可用于环绕安装, 因为任何逃逸幼虫将被卡住和蛹在这条带子上。如果他们这样做, 删除蛹前羽化。</li>
		<li>房子的幼虫和蛹在室温下 (21.5 2.5 °c), 并暴露在间接阳光和房间灯在办公时间或保持在一个光: 黑暗周期的 12 h light:12 h 黑暗。 注: 暴露于24小时的光线会损害存活率。对于从牛场收集的幼虫, 蛹时间从收集后的 1-20 天不等。</li>
		<li>在所需的羽化日期之前, 在悬挂的蚊帐外壳内提供食物和水 (如步骤2.2 所准备), 并每2-3 天更换一次。羽化将在蛹后7-10 天内发生。</li>
		<li>允许新出现的 hoverflies 在室温下喂养6小时, 然后再将它们放进房屋, 如步骤2所述。</li>
	</ol>
</li>
	<li>
妊娠野捉雌性卵的实验室饲养
	<ol>
<li>检查食蚜虻的鸡蛋, 无论是在更换住房之前 (步骤 2), 并在返回到冰箱之前。野生捕获的妊娠雌性产卵发生在8-10 摄氏度和室温。<ol>
<li>将卵放入100毫米 x 20 毫米培养皿中, 含70毫升自来水, 并保持室温直至孵化发生, 通常产卵后2-3 天。</li>
		</ol>
</li>
		<li>将孵出的幼虫放置在2.3 升桶中, 含有1升的新鲜兔粪和1升的自来水。<ol>
<li>每2-3 天检查一次泥浆, 并根据需要添加额外的自来水, 以确保泥浆在 3rd龄幼虫出现之前不会干涸。</li>
		</ol>
</li>
		<li>把水桶放在兔子粪浆中的幼虫, 在一个较大的盒子里 (最小容积30升), 其中含有20升的木屑。确保木屑达到桶边的高度。 注意: 这允许 3rd龄幼虫爬入木屑和蛹。<ol>
<li>将双层蚊帐放在盒子上, 以确保幼虫和任何正在出现的 hoverflies 都不能逃脱。</li>
		</ol>
</li>
		<li>房子的幼虫和蛹在室温下 (21.5 2.5 °c), 并暴露在间接阳光和房间灯在办公时间或保持在一个光: 黑暗周期的 12 h light:12 h 黑暗。 注: 暴露于24小时的光线会损害存活率。</li>
		<li>预计蛹会在15-20 天后发生。收集蛹和放置在昆虫笼, 允许 hoverflies eclose 那里。</li>
		<li>提供食物和水 (按照步骤2.2 中的准备) 在预期的羽化日期之前-羽化将发生在蛹后6-10 天-并更换每2-3 天。</li>
		<li>如步骤2所述, 允许新出现的 hoverflies 在室温下喂养6小时, 然后将它们放进住房。 注意: 幼虫的蛹和蛹的羽化都可以在8-10 摄氏度的黑暗中储存。为此, 将幼虫在兔粪浆和蛹中贮存在木屑中。</li>
	</ol>
</li>
</ol>

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(3-4), 305-314 (1995).</li><li>Hurtado, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Estudio del ciclo de vida de s&#237;rfidos eristalinos (Diptera, Syrphidae) y bases para su cr&#237;a artificial. , (2013).</li><li>Dolley, W. L., White, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+illuminance+on+the+reversal+temperature+in+the+drone+fly,+Eristalis+tenax.">The effect of illuminance on the reversal temperature in the drone fly, Eristalis tenax.</a> Biological Bulletin. 100 (2), 84-89 (1951).</li><li>Wellington, W., Fitzpatrick, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Territoriality+in+the+drone+fly,+Eristalis+tenax+(Diptera,+Syrphidae).">Territoriality in the drone fly, Eristalis tenax (Diptera, Syrphidae).</a> Canadian Entomologist. 113 (6), 695-704 (1981).</li><li>Nordstr&#246;m, K., Barnett, P. D., Moyer de Miguel, I. M., Brinkworth, R. S. A., O'Carroll, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+dimorphism+in+the+hoverfly+motion+vision+pathway.">Sexual dimorphism in the hoverfly motion vision pathway.</a> Current Biology. 18 (9), 661-667 (2008).</li><li>Collett, T. S., Land, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+control+of+flight+behaviour+in+the+hoverfly,+Syritta+pipiens+L.">Visual control of flight behaviour in the hoverfly, Syritta pipiens L.</a> Journal of Comparative Physiology A. 99, 1-66 (1975).</li><li>Pitnick, S., Miller, G., Reagan, J., Holland, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Males'+evolutionary+responses+to+experimental+removal+of+sexual+selection.">Males' evolutionary responses to experimental removal of sexual selection.</a> Proceedings of the Royal Society of London. Series B, Biological Sciences. 268, 1071-1080 (2001).</li><li>Holland, B., Rice, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+removal+of+sexual+selection+reverses+intersexual+antagonists+coevolution+and+removes+a+reproductive+load.">Experimental removal of sexual selection reverses intersexual antagonists coevolution and removes a reproductive load.</a> Proceedings of the National Academy of Sciences USA. 96, 5083-5088 (1999).</li><li>Thyselius, M., Nordstr&#246;m, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hoverfly+locomotor+activity+is+resilient+to+external+influence+and+intrinsic+factors.">Hoverfly locomotor activity is resilient to external influence and intrinsic factors.</a> Journal of Comparative Physiology A. 202 (1), 45-54 (2016).</li><li>Heal, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Colour+patterns+of+Syrphidae:+1.+Genetic+Variation+in+the+dronefly+Eristalis+tenax.">Colour patterns of Syrphidae: 1. Genetic Variation in the dronefly Eristalis tenax.</a> Heredity. 42 (2), 223-236 (1979).</li><li>Ireland, S., Turner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+larval+crowding+and+food+type+on+the+size+and+development+of+the+blowfly,+Calliphora+vomitoria.">The effects of larval crowding and food type on the size and development of the blowfly, Calliphora vomitoria.</a> Forensic Science International. 159, 175-181 (2006).</li><li>Dolley, W. L., Golden, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+sex+and+age+on+the+temperature+at+which+reversal+in+reaction+to+light+in+Eristalis+tenax+occurs.">The effect of sex and age on the temperature at which reversal in reaction to light in Eristalis tenax occurs.</a> Biology Bulletin. 92 (3), 178-186 (1947).</li></ol>

作者没有什么可透露的。

使用我们的技术 (图 1) hoverflies 在实验室中保持了1年的时间, 并在圈养397 月后的行为实验中成功使用。事实上, 即使它似乎违反常理, 保持 hoverflies 在一个更自然的环境, 在12小时 light:12 h 黑暗条件下, 在室温下, 大大降低他们的寿命为2-3 月。在我们的人工休眠周期中维护管超过一年的时间比以前使用不同协议 (77 天33、4月9、18周30) 的尝试要长。影响延长寿命的主要因素可能是人工冬眠的使用量在8-10 摄氏度。通过周期性地打破休眠, 每3-4 天 (步骤 2,图 1), 我们允许 hoverflies 喂养和自新郎, 从而保持营养状态和 hoverflies 的福祉, 这证明了观察到的体重增加 (图形4), 即使在长时间囚禁 (图 3和39) 之后, 也不会改变运动活动。事实上, 关于人工冬眠的失败尝试的文献中的报告并没有打破休眠周期, 从而导致死亡率增加和模具9的存在。
此外, 关于提供花粉作为食物来源, 在整个文献中也存在一些争议。一些论文指出蜂花粉是不够的, 特别是产卵, 只有干燥或确实新鲜的花粉提供适合9,29。我们的发现表明, 通过蜂蜜和水补充蜂花粉, 我们看到的长寿和产卵, 即使经过长时间的圈养, 增加体重在两性 (图 4) 和产卵仍然发生在女性在被囚禁的5月以后在39。延长寿命使我们能够研究 hoverflies 在所有生命阶段的行为。
在田间, 雌性 hoverflies 在季节性冬眠前受精, 并留在生殖滞育期, 精子被储存, 卵母细胞仍未发育, 直到春季28。鉴于一个典型的女性能够在60天内产卵3000个鸡蛋, 因此饲养这些卵是一种快速有效的方法来增加我们圈养的人口.然而, 我们目前对导致卵母细胞在冬眠期后发育的因素的理解是有限的。温度、湿度、光照强度和营养状态都被认为在控制生殖滞育中发挥作用28,40。对这些因素的实验操作可能导致对产卵时间和速率的更大的管理。
同样, 我们已经成功地延缓了幼虫的发育, 以及蛹的羽化, 在黑暗中储藏了2周的8-10 摄氏度, 虽然存活时间可能要长得多。实际上, 治疗30报告了当蛹温度从25摄氏度降至10摄氏度时, 蛹持续时间增加了37天。利用这些战略, 延缓卵子的生产和 (或) 蛹的发展, 可以更大程度地操纵圈养人口的人口统计。
虽然供应的时间一致性对我们的需求比大产量更重要, 但对于其他用途, 例如温室内授粉来说, 这可能更有意义。我们发现, 当使用我们的技术与兔粪便, 我们得到了 163, 34 eclosed hoverflies 从每个离合器的鸡蛋 (N=3)。鉴于一个典型的女性最多可放置200个卵子40, 我们也许能够通过减少过度拥挤和食物竞争, 或者通过调整温度来提高产量, 因为这些都被牵连到对幼虫生长有显著影响的9 ,31,40,41。但是, 没有迹象表明, 媒体的基础对产量的影响有很大的32。此外, 相对于其他脊椎动物的粪便29,30,31,42, 兔粪是比较无异味的, 允许菌落在正常的实验室条件下保存。不需要额外的通风。降低培养基中幼虫的密度, 或添加营养补充剂如酵母, 以及保持恒温20-25 摄氏度之间的恒定温度, 可能足以充分优化产量31,32,40。
收集足够数量的自由飞行 hoverflies, 或维持一个基因多样性的圈养种群的实用性, 对于小型研究项目来说既不切实际, 又有时间限制。因此, 饲养野生交配雌性的后代, 并通过收割成熟的幼虫补充供应品7, 允许在研究设置中为全年使用管提供最实用的选项。由于这些方法受到收集可能发生季节的限制, 因此需要确保成年 hoverflies 的寿命和饲养被捕获的妊娠雌性产卵的任何卵。

Hoverflies 正在成为调查一系列科学问题的有用模型, 包括飞行行为1, 运动视觉的神经机制2, 授粉效率3,4,5,6和微生物免疫7。但是, 与其他一些 dipteran 模型 (如果蝇8) 不同, 在科学研究中没有用于实验室饲养 hoverflies 的标准化协议。事实上, 即使目前的文献描述了育种食蚜虻Eristalis 管的方法, 其中许多都是为大规模种植 hoverflies 作物授粉、生物降解有机废物或解剖学研究而开发的。9,10,11. 因此, 它们没有解决需要一个简单的协议, 提供一致的健康强健的 hoverflies 供应, 同时保持人口的遗传适应性。
跟着蜜蜂和大黄蜂, hoverflies 是最重要的野生, 通才授粉组12,13。全球约有6000种食蚜虻物种, 范围为14, 15, 300 种以上的75属在瑞典16 和300属多个物种在印度 69, 17, 18.例如, 农业上重要的果酱食蚜虻Episyrphus balteatus 和无人机飞行, Eristalis管,, 我们集中在这里, 被发现横跨欧洲, 美国和亚洲6, 16, 17,18,19,20,21,22,23,24,25。Hoverflies 在全年都不一样活跃, 也不全天。事实上, 不仅是季节和白天的时间, 而且还有光强度、温度、湿度和风速的波动, 影响 hoverflies26、27的活动模式。在该字段中, Eristalis可以在一年中的任何时间在地中海气候11中找到, 但在冬季活动的 hoverflies 的数量要低得多。反之, 在寒冷的温带气候下, Eristalis在冬季冬眠, 在10月至 3月28的范围内没有发现积极的行为。
自由飞行 hoverflies 可以通过网在领域收集。事实上, 在温带气候中, 它们是在最丰富的中到晚, 在平静晴朗的日子, 在夏季结束和整个秋季26,27。或者, 成熟的E. 管幼虫, 第二或第三龄, 可以从腐烂的有机物中识别和收获, 如粪便堆或有机污染的溪流10,11。事实上, 为管实验室饲养而发布的技术都是基于在有机污染的水中饲养幼虫, 无论是通过某种形式的植物或粪便物质9,10,29,30,31,32,33. 然而, 幼虫的收集是受季节限制的, 而且是从深秋到秋季11开始的可行的收集工具。此外, 幼虫的丰度受当地天气模式的影响, 因为环境温度的变化会影响产卵和幼虫发育速率的发生,9,28。
因此, 必须通过在实验室内饲养幼虫和卵来维持健康的 hoverflies 种群的战略, 以确保无论季节或当地天气事件, 都能全年进行试验。重要的是, 这里描述的技术孕育了 hoverflies 从野生交配的雌性。这是重要的, 作为研究的 Francuski, et 等。10发现, 一个实验室繁殖的 hoverflies 种群的遗传多样性, 最初是由120成熟的幼虫建立的, 很快就失去了。因此, 他们建议, 为了维持用于商业性作物授粉目的的菌落中的遗传多样性, 这些需要补充, 甚至完全重新建立, 每个春季10都有实地收集的个人。
当研究视觉, 或其他感官使用的求爱和交配, 我们因此建议保持遗传多样性, 无论是重建殖民地或通过补充与野外收集的个人, 定期的殖民地。这一点很重要, 因为性选择会影响到人口的遗传漂移。事实上, 在野外, 雄性 hoverflies 需要识别和拦截合适的配偶, 并通过保卫他们的领地34来与其他雄性争夺交配权利。这一过程确保具有最佳视力和空间关注的男性在交配中可能是最成功的, 因此这些特质将传给下一代。在 hoverflies35、36的视觉通路中存在性 dimorphisms, 部分地证明了这些正在进行的过程的结果。在被囚禁的男性没有同样的障碍, 以成功交配, 如在领域: 首先, 雌性是现成的, 其次, 小的, 封闭的外壳抵消领土行为的影响, 它采取行动, 以阻止其他的交配访问有竞争力的男性。实验性选择清除在果蝇, 已证明对圈养人口有显著的影响, 减少整体体型, 睾丸大小和精子生产37, 并降低男性的比率求爱行为38。因此, 一个圈养繁殖计划, 不考虑性选择, 可能会对随后进行的视觉和行为研究产生深远的影响。
我们在这里描述一个简单和经济高效的解决方案, 提供了一致的健康 hoverflies 供应。该协议灵活, 易于重新启动和/或高级, 取决于研究的要求。

<table border="0" cellpadding="0" cellspacing="0" style="width:903px;" width="902">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Bee Pollen</td>
			<td style="width:211px;">Forest Super Foods</td>
			<td style="width:119px;"></td>
			<td style="width:357px;">any brand of bee pollen is suitable</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Honey</td>
			<td style="width:211px;">Bramwells</td>
			<td style="width:119px;"></td>
			<td>any brand of liquid honey is suitable</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Rabbit Faeces</td>
			<td style="width:211px;"></td>
			<td style="width:119px;"></td>
			<td>can be substituted with cow or pig manure made into a slurry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">BugDome</td>
			<td style="width:211px;">Australia Entomological Supplies</td>
			<td style="width:119px;">EM42222</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Plastic Bags</td>
			<td style="width:211px;">Woolworths Homebrand</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mosquito netting</td>
			<td style="width:211px;">Clas Ohlson</td>
			<td>34-1113</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cotton Balls</td>
			<td style="width:211px;">Woolworths Select</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fridge</td>
			<td>Hisense</td>
			<td style="width:119px;"></td>
			<td>fridge needs to maintain a stable 8-10&#176;C&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Buckets (2-3L)</td>
			<td style="width:211px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Large plastic tubs (30L)</td>
			<td style="width:211px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Wood shavings</td>
			<td style="width:211px;">Pollards Sawdust Supplies</td>
			<td>MaxiFlake (75)&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Bag clips</td>
			<td style="width:211px;">IKEA</td>
			<td style="width:119px;">Bevara 303.391.70</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Petri Dish (100mm x 20mm)</td>
			<td style="width:211px;">Corning</td>
			<td style="width:119px;">430167</td>
			<td></td>
		</tr>
	</tbody>
</table>

rearing and long-term maintenance of eristalis tenax hoverflies for research studies

在全世界估计有6000种物种, hoverflies 在生态学上是重要的, 作为替代传粉者的驯化蜜蜂。然而, 它们也是一个有用的科学模型来研究运动视觉和飞行动力学在一个控制的实验室设置。随着幼虫在有机污染水体中的发育, 它们是研究微生物免疫投资的有用模型。虽然大规模的农业商业育种已经发生, 但没有标准化的议定书来维持圈养的人口进行科学研究。这一点很重要, 因为在高峰授粉期间, 专注于大规模产出的商业性圈养育种计划可能无法提供全年一致、稳定和健壮的人口, 因为其他研究目的通常都需要这样做。因此, 需要建立、维持和更新圈养研究人口的方法。在这里, 我们描述了一个人工休眠循环的使用, 除了营养和住房需求, 长期维护Eristalis 管。使用这些方法, 与以前的报告相比, 我们大大提高了管中的捕获种群的健康和寿命。此外, 我们还讨论了小规模饲养方法和选择, 以优化产量和操纵人口统计。

我们已经制定了一个三路策略, 它为视觉和行为研究保持健康的人口 (在图 1中总结)。我们的方法从野外收集 hoverflies (步骤1、图 1) 开始。在我们的实验室中, hoverflies 在人工休眠周期 (步骤2、图 1) 下存放在昆虫笼或塑料袋中, 大大延长了它们的寿命。对于增加的数字, 后代可以从野生交配的雌性饲养 (步骤 3,图 1)。
我们发现, 捕捉大量的野生 hoverflies 是一个时间密集型的努力, 即使环境条件是有利的。相比之下, 从牛场的粪便中收获的成熟幼虫的成功饲养是一种更有效的方式来来源大量野生 hoverflies (步骤 1,图 1), 我们收集多达700幼虫在0.03 米3粪。此外, 我们的技术, 由捕获的妊娠雌性奠定的后卵已经证明是成功的 (步骤 3,图 1)。在秋季和冬季, 在地中海气候 (阿德莱德) 捕获的雌性产卵了几批卵, 在20周的时间里, 有24组从19只雌性中观察到。在这些卵批次中, 10 被放入水中, 所有这些都是肥沃的, 导致幼虫孵化。3组幼虫然后被采取在这一点之外, 并且安置在兔子粪浆, 导致 163 N=3 34 (意味的) 出现了 hoverflies, 没有被观察的性别偏见 (图 2). 
这些实验室饲养 hoverflies 的健康状况是通过比较女性 hoverflies 的体重和运动活动与野外捕获的个体相比而确定的。一般运动活动的评估使用运动活动监测系统 (LAMS), 如前所述39。在人工休眠周期内, 在圈养4月后, 在实验室饲养和野生捕获 hoverflies 之间, 观察到在饲养和野外捕捞后的3间, 在体重方面没有显著差异 (图) 或活动 (图 3B)。当管在实验室中保持不使用人工休眠周期时, 我们看到寿命显著下降, 其寿命为 2.5-3 月 (5 名女性 73 @ 7 天, 79 个男性4天)。当 hoverflies 被保存在人工冬眠时, 他们可以活超过12月。
此外, 使用我们描述的方法, 长期维护的影响, 进一步评估的重量随着时间的推移, 对实验室饲养 hoverflies 的性别。我们观察到, 男女在4月的时间里体重显著增加, 女性比男性更重 (p < 0.0001, 双向方差分析, N=12,图 4)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57711/57711fig1.jpg"> 图形1:概述维护健康圈养人口的方法的流程图管.(1) 此处描述的过程开始于从牛粪 (步骤 1.2) 或自由飞行 hoverflies (步骤 1.3) 中收集成熟幼虫。(2) hoverflies 被安置在昆虫笼或塑料袋中, 依赖于数字。他们被保存在一个人工冬眠周期在8-10 °c, 每3-4 天被打破。(3) 收集的幼虫保存在牛粪中 (步骤 3.2)。在实验室放置的鸡蛋放在兔子粪浆中 (步骤 3.3)。当达到成熟度时, 3 个rd龄幼虫爬入周围的蛹的灰尘。羽化发生在6-10 天后, eclosed hoverflies 放置在外壳 (步骤 2) 中。<a href="https://www.jove.com/files/ftp_upload/57711/57711fig1large.jpg">请单击此处下载此文件.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57711/57711fig2v3.jpg"> 图形2:编号和性别比率管成功地从单个鸡蛋批次中饲养.  数据显示管的数目, eclosed 从蛹中开发出的3批鸡蛋。这些卵是由野生捕获的雌性产卵的。数据是彩色编码的苍蝇的性别。这一比例没有显著差异。
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57711/57711fig3v3.jpg"> 图形3:长期  圈养后在实验室饲养和田间采集的女性 hoverflies 中对人口健康的评估。(A)人工冬眠 (N=12) 后4月内饲养的实验室和野外捕获的雌性 hoverflies 的体重比较。(B)在人工冬眠下圈养4月后, 实验室饲养的和野外捕获的雌性 hoverflies 的活动水平。在运动过程中, 通过对一束红外光束的破坏, 测量了 hoverflies 的活动活动系统。如前所述, 我们在一天中平均6.7 小时的活动, 在运动活动监测系统的第二个完整的一天,39 (n字段捕获= 9, n实验室饲养= 12)。每个箱线图的中心标记显示了中间值、框的边缘25 th 到75 th 百分点的数据, 而胡须从最小到数据的最大值。
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57711/57711fig4v3.jpg"> 图形4:长期维护对实验室饲养的 hoverflies 的重量的影响比较.  数据显示, 食蚜虻重量是在人工冬眠条件下囚禁时间的函数。由于所有的 hoverflies 都是从我们实验室的卵 (N=12 在每个数据点) 饲养的, t=0 等于蛹孵化时间, 圈养的时间与动物的年龄相同。每个箱线图的中心标记显示了中间值、框的边缘25 th 到75th 百分点的数据, 而胡须从最小到数据的最大值。

Watch this Scientific Journal Video about Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies at JoVE.com

Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

研究性学习的Eristalis 管Hoverflies 的饲养和长期维护

这些过程的总体目标是在研究设置中建立、维护和刷新Eristalis 管的俘虏群体。

With an estimated 6000 species worldwide, hoverflies are ecologically important as alternative pollinators to domesticated honeybees. However, they are ...

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Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

13.3K 次观看.  Flinders University. 这些过程的总体目标是在研究设置中建立、维护和刷新Eristalis 管的俘虏群体。

视频: Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

Waste Water Derived Electroactive Microbial Biofilms: Growth, Maintenance, and Basic Characterization

The growth of anodic electroactive microbial biofilms from waste water inocula in a fed-batch reactor is demonstrated using a three-electrode setup controlled by a potentiostat. Thereby the use of potentiostats allows an exact adjustment of the electrode potential and ensures reproducible microbial culturing conditions. During growth the current production is monitored using chronoamperometry (CA). Based on these data the maximum current density (jmax) and the coulombic efficiency (CE) are discussed as measures for characterization of the bioelectrocatalytic performance. Cyclic voltammetry (CV), a nondestructive, i.e. noninvasive, method, is used to study the extracellular electron transfer (EET) of electroactive bacteria. CV measurements are performed on anodic biofilm electrodes in the presence of the microbial substrate, i.e. turnover conditions, and in the absence of the substrate, i.e. nonturnover conditions, using different scan rates. Subsequently, data analysis is exemplified and fundamental thermodynamic parameters of the microbial EET are derived and explained: peak potential (Ep), peak current density (jp), formal potential (Ef) and peak separation (&#916;Ep). Additionally the limits of the method and the state-of the art data analysis are addressed. Thereby this video-article shall provide a guide for the basic experimental steps and the fundamental data analysis.

Chronoamperometric growth of anodic electroactive microbial biofilms in a fed-batch reactor using a three-electrode setup, controlled by a potentiostat, is demonstrated. The extracellular electron transfer is characterized using cyclic voltammetry in the presence of the electron donor and in the absence of the electron donor. Fundamental data analysis is demonstrated.

The growth of anodic electroactive microbial biofilms from waste water inocula in a fed-batch reactor is demonstrated using a three-electrode setup ...

科研

环境科学

Design and Construction of an Urban Runoff Research Facility

As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use due to increased demand for landscape irrigation. Improper irrigation practices and large rainfall events can result in runoff from urban landscapes which has potential to carry nutrients and sediments into local streams and lakes where they may contribute to eutrophication. A 1,000 m2 facility was constructed which consists of 24 individual 33.6 m2 field plots, each equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated urban landscapes. Runoff volumes from the first and second trials had coefficient of variability (CV) values of 38.2 and 28.7%, respectively. CV values for runoff pH, EC, and Na concentration for both trials were all under 10%. Concentrations of DOC, TDN, DON, PO4-P, K+, Mg2+, and Ca2+ had CV values less than 50% in both trials. Overall, the results of testing performed after sod installation at the facility indicated good uniformity between plots for runoff volumes and chemical constituents. The large plot size is sufficient to include much of the natural variability and therefore provides better simulation of urban landscape ecosystems.

This paper describes the design, construction, and function of a 1,000 m2 facility containing 24 individual 33.6 m2 field plots equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated home lawns.

设计一个城市径流研究设施的建设

As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use ...

Physical, Chemical and Biological Characterization of Six Biochars Produced for the Remediation of Contaminated Sites

The physical and chemical properties of biochar vary based on feedstock sources and production conditions, making it possible to engineer biochars with specific functions (e.g. carbon sequestration, soil quality improvements, or contaminant sorption). In 2013, the International Biochar Initiative (IBI) made publically available their Standardized Product Definition and Product Testing Guidelines (Version 1.1) which set standards for physical and chemical characteristics for biochar. Six biochars made from three different feedstocks and at two temperatures were analyzed for characteristics related to their use as a soil amendment. The protocol describes analyses of the feedstocks and biochars and includes: cation exchange capacity (CEC), specific surface area (SSA), organic carbon (OC) and moisture percentage, pH, particle size distribution, and proximate and ultimate analysis. Also described in the protocol are the analyses of the feedstocks and biochars for contaminants including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), metals and mercury as well as nutrients (phosphorous, nitrite and nitrate and ammonium as nitrogen). The protocol also includes the biological testing procedures, earthworm avoidance and germination assays. Based on the quality assurance / quality control (QA/QC) results of blanks, duplicates, standards and reference materials, all methods were determined adequate for use with biochar and feedstock materials. All biochars and feedstocks were well within the criterion set by the IBI and there were little differences among biochars, except in the case of the biochar produced from construction waste materials. This biochar (referred to as Old biochar) was determined to have elevated levels of arsenic, chromium, copper, and lead, and failed the earthworm avoidance and germination assays. Based on these results, Old biochar would not be appropriate for use as a soil amendment for carbon sequestration, substrate quality improvements or remediation.

Biochar is a carbon-rich material used as a soil amendment with the ability to sustainably sequester carbon, improve substrate quality and sorb contaminants. This protocol describes the 17 analytical methods used for the characterization of biochar, which is required prior to large scale implementation of these amendments in the environment.

物理，化学和六Biochars生产的生物表征污染场地修复的

The physical and chemical properties of biochar vary based on feedstock sources and production conditions, making it possible to engineer biochars with ...

Administering and Detecting Protein Marks on Arthropods for Dispersal Research

Monitoring arthropod movement is often required to better understand associated population dynamics, dispersal patterns, host plant preferences, and other ecological interactions. Arthropods are usually tracked in nature by tagging them with a unique mark and then re-collecting them over time and space to determine their dispersal capabilities. In addition to actual physical tags, such as colored dust or paint, various types of proteins have proven very effective for marking arthropods for ecological research. Proteins can be administered internally and/or externally. The proteins can then be detected on recaptured arthropods with a protein-specific enzyme-linked immunosorbent assay (ELISA). Here we describe protocols for externally and internally tagging arthropods with protein. Two simple experimental examples are demonstrated: (1) an internal protein mark introduced to an insect by providing a protein-enriched diet and (2) an external protein mark topically applied to an insect using a medical nebulizer. We then relate a step-by-step guide of the sandwich and indirect ELISA methods used to detect protein marks on the insects. In this demonstration, various aspects of the acquisition and detection of protein markers on arthropods for mark-release-recapture, mark-capture, and self-mark-capture types of research are discussed, along with the various ways that the immunomarking procedure has been adapted to suit a wide variety of research objectives.

Proteins are often used to mark arthropods for dispersal research. Methods for administering internal and external protein marks on arthropods are demonstrated. Protein mark detection techniques, either through indirect or sandwich enzyme-linked immunosorbent assays (ELISAs), are also illustrated.

管理和检测蛋白标记的节肢动物的扩散研究

Monitoring arthropod movement is often required to better understand associated population dynamics, dispersal patterns, host plant preferences, and other ...

Clean Sampling and Analysis of River and Estuarine Waters for Trace Metal Studies

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination control. Developments of some techniques aiming to reduce trace metal contamination in the last couple of decades have resulted in concentrations reported now being orders of magnitude lower than those in the past. These low concentrations often necessitate preconcentration of water samples prior to instrumental analysis of samples. Since contamination can appear in all phases of trace metal analyses, including sample collection (and during preparation of sampling containers), storage and handling, pretreatments, and instrumental analysis, specific care needs to be taken in order to reduce contamination levels at all steps. The effort to develop and utilize "clean techniques" in trace metal studies allows scientists to investigate trace metal distributions and chemical and biological behavior in greater details. This advancement also provides the required accuracy and precision of trace metal data allowing for environmental conditions to be related to trace metal concentrations in aquatic environments.
This protocol that is presented here details needed materials for sample preparation, sample collection, sample pretreatment including preconcentration, and instrumental analysis. By reducing contamination throughout all phases mentioned above for trace metal analysis, much lower detection limits and thus accuracy can be achieved. The effectiveness of "clean techniques" is further demonstrated using low field blanks and good recoveries for standard reference material. The data quality that can be obtained thus enables the assessment of trace metal distributions and their relationships to environmental parameters.

Special care using "clean techniques" is required to properly collect and process water samples for trace metal studies in aquatic environments. A protocol for sampling, processing, and analytical procedures with the aim of obtaining reliable environmental monitoring data and results with high sensitivity for detailed trace metal studies is presented.

清洁取样和痕量金属研究河和河口水域分析

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination ...

Use of a Filter Cartridge for Filtration of Water Samples and Extraction of Environmental DNA

Recent studies demonstrated the use of environmental DNA (eDNA) from fishes to be appropriate as a non-invasive monitoring tool. Most of these studies employed disk fiber filters to collect eDNA from water samples, although a number of microbial studies in aquatic environments have employed filter cartridges, because the cartridge has the advantage of accommodating large water volumes and of overall ease of use. Here we provide a protocol for filtration of water samples using the filter cartridge and extraction of eDNA from the filter without having to cut open the housing. The main portions of this protocol consists of 1) filtration of water samples (water volumes &#8804;4 L or &gt;4 L); (2) extraction of DNA on the filter using a roller shaker placed in a preheated incubator; and (3) purification of DNA using a commercial kit. With the use of this and previously-used protocols, we perform metabarcoding analysis of eDNA taken from a huge aquarium tank (7,500 m3) with known species composition, and show the number of detected species per library from the two protocols as the representative results. This protocol has been developed for metabarcoding eDNA from fishes, but is also applicable to eDNA from other organisms.

We describe a protocol for filtration of water samples with a filter cartridge and extraction of environmental DNA (eDNA) without having to cut open the housing to remove the filter. This protocol is developed for metabarcoding eDNA from fishes, but is also applicable to eDNA from other organisms.

滤芯水过滤样品和环境DNA的提取使用

Recent studies demonstrated the use of environmental DNA (eDNA) from fishes to be appropriate as a non-invasive monitoring tool. Most of these studies ...

RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop cultivars against environmental stresses. An experimental protocol is presented for RGB and hyperspectral imaging of root systems. The approach uses rhizoboxes where plants grow in natural soil over a longer time to observe fully developed root systems. Experimental settings are exemplified for assessing rhizobox plants under water stress and studying the role of roots. An RGB imaging setup is described for cheap and quick quantification of root development over time. Hyperspectral imaging improves root segmentation from the soil background compared to RGB color based thresholding. The particular strength of hyperspectral imaging is the acquisition of chemometric information on the root-soil system for functional understanding. This is demonstrated with high resolution water content mapping. Spectral imaging however is more complex in image acquisition, processing and analysis compared to the RGB approach. A combination of both methods can optimize a comprehensive assessment of the root system. Application examples integrating root and aboveground traits are given for the context of plant phenotyping and plant physiological research. Further improvement of root imaging can be obtained by optimizing RGB image quality with better illumination using different light sources and by extension of image analysis methods to infer on root zone properties from spectral data.

An experimental protocol is presented for assessment of soil grown plant root systems with RGB and hyperspectral imaging. Combination of RGB image time series with chemometric information from hyperspectral scans optimizes insights into plant root dynamics.

RGB 和光谱根成像植物表型和生理研究: 实验装置和成像协议

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop ...

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 ...

Long-term Video Tracking of Cohoused Aquatic Animals: A Case Study of the Daily Locomotor Activity of the Norway Lobster (Nephrops norvegicus)

We present a protocol related to a video-tracking technique based on the background subtraction and image thresholding that makes it possible to individually track cohoused animals. We tested the tracking routine with four cohoused Norway lobsters (Nephrops norvegicus) under light-darkness conditions for 5 days. The lobsters had been individually tagged. The experimental setup and the tracking techniques used are entirely based on the open source software. The comparison of the tracking output with a manual detection indicates that the lobsters were correctly detected 69% of the times. Among the correctly detected lobsters, their individual tags were correctly identified 89.5% of the times. Considering the frame rate used in the protocol and the movement rate of lobsters, the performance of the video tracking has a good quality, and the representative results support the validity of the protocol in producing valuable data for research needs (individual space occupancy or locomotor activity patterns). The protocol presented here can be easily customized and is, hence, transferable to other species where the individual tracking of specimens in a group can be valuable for answering research questions.

Long-term Video Tracking of Cohoused Aquatic Animals: A Case Study of the Daily Locomotor Activity of the Norway Lobster Nephrops norvegicus

Here we present a protocol to individually track animals over a long period of time. It uses computer vision methods to identify a set of manually constructed tags by using a group of lobsters as case study, simultaneously providing information on how to house, manipulate, and mark the lobsters.