This research was funded by Washington College's Provost's Office, Middendorf Fund, Hodson Trust, and Franklin and Marshall's Hackman Fund and College of Grants. We thank E. Counihan, S. Giordano, F. Rauh, and A. Roth for assistance in the field. We thank M. Conner, R. Fleegle, and D. Startt at Chesapeake Farms, and Chino Farms for permission and access. The Washington College GIS Program helped with the preparation of maps.

所有涉及动物对象的程序是由富兰克林和马歇尔和华盛顿学院的机构动物护理和使用委员会批准，并遵循所有当地，州和联邦法规。 1.捕获和处理<ol><li>放置箍陷阱的该已知含有龟水目标体。确定水深确保4 - 陷阱5英寸仍高于水，让被困的海龟浮出水面呼吸。可以肯定，扩大和保护圈陷阱，其最大长度与横梁，防止陷阱崩溃。桩陷阱入水体的床，以防止漂移。 </li><li>诱饵的陷阱与一些死鱼，鸡肝，或鸡脖子，猫粮罐头，和/或蔬菜25即可。 </li><li>检查陷阱，每天两次，并删除海龟。当从陷阱取出乌龟，由边抱动，谨慎使用，以避免爪子或喙伤害。 <ol><li>副渔获物的释放。回覆若继续俘获需要设置陷阱。评估诱饵的状态。如果诱饵已被消耗，增加更多。拉陷阱，如果附加诱捕不希望到岸边。 </li></ol></li><li>如果需要26，27决定龟的性别和年龄。 </li><li>将龟在保持袋，并测量体重用弹簧秤到最近的克</li><li>运输龟在气候受控运输箱在实验室用的少量水。房子单独的动物中，与未处理池塘水水族馆在25℃左右举行，深足只盖住头部。 </li></ol> 2.粘贴无线发射机<ol><li>为了最大限度地提高发射机寿命和输出，选择最大的无线电发射机可能不超过该动物的体重28的5％。 </li><li>大约一半确定发射器放置的位置上背甲背甲中线和横向边缘之间大约升1/3从背甲的后缘ength起来。 </li><li>用干布去除淤泥，杂物和藻类生长准备区。用70％异丙醇拭区。 </li><li>用5分钟环氧树脂少量附加发射机。定向发射机以最大化与甲壳的表面接触。使得它落后平行于主体的长轴动物后面的天线的位置。 </li><li>一旦适当地定位，覆盖整个发射器，并用5分钟环氧约为1厘米甲壳周围的表面。 </li><li>返回龟水族馆，让环氧树脂固化过夜。 </li></ol> 3.药理准备注意:氢溴酸东莨菪碱和东莨菪碱甲基溴是强效的乙酰胆碱受体拮抗剂。当使用这些药物的工作，请查阅材料安全数据表，使用适当的个人防护装备（ 如手套，通风橱），并遵守实验室的安全协议，以避免accidenTAL接触。 <ol><li>使用无菌和antipyrogenic用品，化合物氢溴酸东莨菪碱的储备溶液。称出药物的所需量在分析天平上。混合用注射盐水的东莨菪碱在锥形管的1毫克/毫升的浓度。涡旋溶液直至溶解。确保基的化学品是化学纯度符合或超过美国药典（USP）制剂时可能11。 </li><li>重复步骤3.1与东莨菪碱甲基溴化物。 </li><li>通过0.22微米孔径的尼龙或混合纤维素酯处理溶液注射器过滤器进入无菌密封的血清小瓶中。 </li><li>室温保存。 24小时内使用。 </li></ol> 4.追踪海龟运动使用无线电遥测11，12 <ol><li>移动到目标动物的大致位置，从动物其余至少25米使用定向八木天线和设置在中等接收机增益，扫描卧式上，以确定动物的粗方向和位置。当接收干扰或弱信号，找到了新的位置。增加抬高或持有八木高空时，可以提高信号。 </li><li>一旦一个合适的位置被发现采取轴承，记录与GPS你的立场。 </li><li>使用空/峰值方法28，确定左和右的空值的轴承。 <ol><li>识别最强信号的方向。转动增益向下尽可能同时仍接收可检测信号。在接收器上使用衰减开关，如果这样装备。移动天线向左侧并记录在该信号丢失的罗盘方位。 </li><li>重复正确的轴承上一步。 </li></ol></li><li>重复步骤4.3从同一动物的至少两个附加位置。 注:这些附加分应该采取这样的方式，以包围所述动物。 <ol><li>当一组轴承是从格雷亚取ŧ距离或不可避免的干扰，至少需要两个额外的轴承，以提高准确性。收集套用于尽可能快地估算一个位置，特别是如果动物移动的轴承。 注:此外，多个人员可以协调就可以同时在同一动物的多个独立的轴承。 </li></ol></li><li>使用上述步骤15分钟 - 数字或手工每10动物的记录位置。 </li><li>认知过程的操纵<ol><li>一旦未经处理动物的路径记录（一个主体内的控制），提供了一个既剂量的东莨菪碱或甲基东的。使用在步骤1.5中收集的质量，计算药物的量给予动物以达到6.4毫克/千克东莨菪碱或6.8毫克/千克甲基东19,20的一个特定的爬行动物的最终剂量。 </li><li> 通过使用1.0M的尾腹膜窦直接递送药物进入腹膜升注射器用22号针头。确保输送的总体积不超过1毫升</li><li>在捕获其网站上尽快释放动物。 </li><li>继续监测动物的运动，每10 - 15分钟，直到它到达其预期目的地。 </li></ol></li></ol> 5.空间分析<ol><li>计算动物的位置的估计。 <ol><li>对于每一组的空轴承，平分角（用手或通过软件），以找到所得发射机轴承。重复在给定时间点的所有轴承。 </li><li>根据制造商的协议使用三角测量软件，估计动物的与多个发射机的轴承位置和相关联的错误。转换位置估计为x / y坐标，如果软件提供不同的输出。 </li></ol></li><li>重复步骤5.1轴承所有集合。 </li><li>计算运动的空间精度。 <ol><li>计算累积的几何平均值未经处理的动物（阴性对照），因为他们对自己的目标12移动（ 即平均路径）。 </li><li>用于治疗和阳性对照组，每组个体，计算直到所有的点的100％已列入11,12重叠来自5个米的间隔的几何平均线依次较大幅点的累积数目。 </li><li>计算的平均值和标准误差在每个组中的所有个体的每个分割行的距离。 </li><li>统计分析的数据时，治疗组的时间间隔中任一用于比较在给定的时间间隔，以容纳点或重叠点的100％，这取决于问题。 </li></ol></li></ol>

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The authors have nothing to disclose.

这里介绍的协议允许实验者记录和量化在导航认知的作用。在现场操作的认知已经证明是困难的，因为多数的方法发表实验者无法知道哪些动物的行为的具体方面是被操纵。然而，这里提出的协议允许实验者准确操纵，从而评估在导航认知的作用。该技术还允许实验者监测实时动物导航与异常高的空间和时间分辨率，由此赋予研究者清楚地记录在野生动物的认知实验操作的行为后果。 <p class="j...

认知（本文中定义为"涉及采集，存储，并使用从环境信息的所有进程"1）是中央的复杂的导航任务2的阵列。例如，沙丘鹤（ 丹顶鹤加拿大一枝黄花 ），显示精度迁徙对他们的本命的海滩幼体有明显改善与经验; 3，和海龟物种的印记，并返回作为成年人4-6。同样，成功的迁移，扩散和觅食的动物的能力，铰链，以收集有关其空间环境7,8的信息。有些动物出现学习就具体的景观特色9航行路线和筑巢和觅食区10之间移动时，可以使用空间认知。在东部锦龟最近的工作（Chrysemys PICTA）建议在航海，其中成人高地栖息地的成功导航尤文铰链的关键时期一个狭窄的年龄范围（&lt;4岁11-13）内Nile体验。虽然这些一起研究表明，在了解航海4-6，14-16学习的作用所取得的进展，背后这些行为的机制和认知导航的充分作用仍然是谜一般的，尤其是在脊椎动物8，17 18。 实地调查认知导航的作用是罕见的2，8，18，这主要是由于参与监督的方法上的困难，处理和跟踪野生动物。例如，大空间和时间尺度上许多动物进行导航往往妨碍调查两种类型的信息，这些动物可能学习和信息是如何获得的。实验者经常面对的检测和监控行为在如此大面积和时间框架时定位的动物，从而限制了类型的后勤困难的，可以被收集的数据，并且可以得出的结论。虽然使用动物安装全球定位系统（GPS）的记录器可以提高检测广泛范围的动物的概率，通过这些装置收集的空间数据一般都非常粗分辨率的和缺乏详细的行为的组件。因此，能在这种情况下要收集的数据是用于在行为检查不同的组或实验组之间微妙的变化价值有限。同样地，目标行为的直接受控操纵通常是由典型的导航行为，以及由实地研究的固有后勤限制的空间和时间尺度禁止。发现动物的自然栖息地，捕捉和操纵它们，然后收集行为数据，而无需在不经意间产生的寄生行为在外地工作动物的重大挑战。因此，对FR实验设计EE-不等的动物往往是制约和对认知的导航作用进行严格，控制现场实验的能力是有限的。 本研究绕过许多用在野外条件下的新的药理学处理和自由导航动物高分辨率跟踪的组合调查在该领域认知和导航之间的关系的先前困难。东莨菪碱，毒蕈碱乙酰胆碱受体（mAchR的）拮抗剂，已显示在多种脊椎动物类群18-24的脑中阻断胆碱能活性阻断空间记忆形成和召回。东莨菪碱能有效地对自由放养动物的野外条件下11，18下使用并有标记，但暂时的效果（ 例如 ，6 -爬行动物8小时）。甲基东，即不穿过血脑屏障19-21一个mAchR的拮抗剂，可用于控制东莨菪碱和行为11的非认知方面的可能外周作用。药理学允许认知的精确操纵通过直接影响受体，和高精度的无线电遥测允许的对行为所得效果的观察。 通过远程三角同时具有高的空间（±2.5米）和时间（15分钟）的分辨率进行的测量允许精确记录和相对动物行为的量化，以认知的实验操作。 这项研究11切萨皮克农场，在肯特公司，MD一个3300英亩野生动物管理和农业研究领域，美国（39.194°N，76.187°W）五月和2013年8月和2014年间进行的。该协议包括五个主要步骤:（1）捕捉和处理的动物（2）固定用无线电发射机（3）制备的药理学试剂（4）监测和操纵动物的动作，和（5）ANAlyzing空间数据。本文所述集中在东部的研究锦龟（Chrysemys PICTA）。在重点人群海龟参与其中，他们离开自己的家的池塘，然后导航到使用四种非常精确的（±3.5米）的，复杂的，高度可预测的路线11，12替代水生境的年度陆上运动。在动物药理操纵该系统具有高分辨率无线电遥测配对揭示认知的自由航行野生动物的角色光芒。

<table border="0" cellpadding="0" cellspacing="0" width="595">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Scopolamine bromide</td>
			<td style="width:93px;">Sigma</td>
			<td style="width:119px;">S0929</td>
			<td style="width:167px;">USP</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;">Scopolamine methylbromide</td>
			<td>Sigma</td>
			<td>S8502, 1421009</td>
			<td style="width:167px;">USP and non USP versions</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;">Saline</td>
			<td>Hanna Pharmaceutical Supply Co., Inc.</td>
			<td>409488850</td>
			<td style="width:167px;">USP, formulated as an injectable&#160;</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Syringe filter</td>
			<td>Fisher</td>
			<td>09-720-004</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Syringe</td>
			<td>Fisher</td>
			<td>14-823-30</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Hypodermic needle</td>
			<td>Fisher</td>
			<td>14-823-13</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;">Antenna</td>
			<td>Wildlife Materials</td>
			<td>3 Element Folding Yagi</td>
			<td style="width:167px;">Antennae with additional elements are available, but can be cumbersome in the field.&#160;</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;">Radio Receiver</td>
			<td>Wildlife Materials</td>
			<td>TRX-2000S</td>
			<td style="width:167px;">Water resistant models are also available.</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Compass</td>
			<td>Brunton&#160;</td>
			<td>Truarc 15</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="169">
			<td height="169" style="height:169px;">Radio transmitters</td>
			<td>Holohil Inc.</td>
			<td>BD-2, PD-2, RI-2B</td>
			<td style="width:167px;">Transmitter models vary in lifespan and signal output as a function of battery size and pulse rate settings, which can be customized based on the study question and organism.</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">GPS</td>
			<td>Garmin</td>
			<td>eTrex Venture</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;">Coaxial cable</td>
			<td>newegg.com</td>
			<td>C2G 40026</td>
			<td style="width:167px;">BNC connections are necessary.</td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;">Hoop net</td>
			<td>Memphis Net and Twine&#160;</td>
			<td>TN325</td>
			<td style="width:167px;">Net mesh size should be chosen based on the minimum size of the target animal.&#160;</td>
		</tr>
	</tbody>
</table>

using pharmacological manipulation and high-precision radio telemetry to study the spatial cognition in free-ranging animals

动物的感知和了解其环境的能力起着许多行为过程，包括导航，迁移，扩散和觅食了关键作用。然而，认知的导航策略和这些策略背后的机制发展中的作用的认识是通过参与监督的方法上的困难限制，操纵的认知和跟踪野生动物。这项研究描述为解决认知导航，它结合了高精度无线电遥测行为药理操纵角色的协议。该方法使用东莨菪碱，毒蕈碱乙酰胆碱受体拮抗剂，操纵认知空间能力。处理的动物中，然后用高频率和通过远程三角高空间分辨率监测。该协议是一个人口的东锦龟（Chrysemys PICTA）内的应用已经有人居住季节性短暂的水源〜100年间，采用精确的（±3.5米）远源之间移动，复杂的（ 即具有较高的曲折经过多个栖息地非线性），且可预测的路线前4岁学会了。这项研究表明，由这些龟所使用的过程是与空间记忆形成和召回一致。总之，这些结果与空间认知的复杂的导航作用是一致的，突出的生态和药理学技术认知和导航的研究的整合。

采用上述方案，认知导航的作用，东部人口进行评估锦龟（Chrysemys PICTA）已经经历了〜百年的季节性短暂的水源。这一人群栖息的短暂的混合（每年并迅速排出-在几个小时）和永久水生生境（ 图1）。先前的研究表明，它们的池塘排水后，居民龟导航到以高精度（±3.5米）在使用前4岁11-13（ 图1）了解到复杂的，可预测的路线的替代...

Watch this Scientific Journal Video about Using Pharmacological Manipulation and High-precision Radio Telemetry to Study the Spatial Cognition in Free-ranging Animals at JoVE.com

Using Pharmacological Manipulation and High-precision Radio Telemetry to Study the Spatial Cognition in Free-ranging Animals

本文介绍了结合内存和无线电遥测的药理操作记录和量化导航认知的作用新颖的协议。

使用药理操纵和高精度无线电遥测研究空间认知中自由放养的动物

An animal&#39;s ability to perceive and learn about its environment plays a key role in many behavioral processes, including navigation, migration, ...

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Research

JoVE Journal

Environment

6.7K 次观看.  Franklin and Marshall College. 本文介绍了结合内存和无线电遥测的药理操作记录和量化导航认知的作用新颖的协议。 

视频: Using Pharmacological Manipulation and High-precision Radio Telemetry to Study the Spatial Cognition in Free-ranging Animals

The Use of an Automated System (GreenFeed) to Monitor Enteric Methane and Carbon Dioxide Emissions from Ruminant Animals

Ruminant animals (domesticated or wild) emit methane (CH4) through enteric fermentation in their digestive tract and from decomposition of manure during storage. These processes are the major sources of greenhouse gas (GHG) emissions from animal production systems. Techniques for measuring enteric CH4 vary from direct measurements (respiration chambers, which are highly accurate, but with limited applicability) to various indirect methods (sniffers, laser technology, which are practical, but with variable accuracy). The sulfur hexafluoride (SF6)&#160;tracer gas method is commonly used to measure enteric CH4 production by animal scientists and more recently, application of an Automated Head-Chamber System (AHCS) (GreenFeed, C-Lock, Inc., Rapid City, SD), which is the focus of this experiment, has been growing. AHCS is an automated system to monitor CH4 and carbon dioxide (CO2) mass fluxes from the breath of ruminant animals. In a typical AHCS operation, small quantities of baiting feed are dispensed to individual animals to lure them to AHCS multiple times daily. As the animal visits AHCS, a fan system pulls air past the animal&#8217;s muzzle into an intake manifold, and through an air collection pipe where continuous airflow rates are measured. A sub-sample of air is pumped out of the pipe into non-dispersive infra-red sensors for continuous measurement of CH4 and CO2 concentrations. Field comparisons of AHCS to respiration chambers or SF6 have demonstrated that AHCS produces repeatable and accurate CH4 emission results, provided that animal visits to AHCS are sufficient so emission estimates are representative of the diurnal rhythm of rumen gas production. Here, we demonstrate the use of AHCS to measure CO2 and CH4 fluxes from dairy cows given a control diet or a diet supplemented with technical-grade cashew nut shell liquid.

The Use of an Automated System GreenFeed to Monitor Enteric Methane and Carbon Dioxide Emissions from Ruminant Animals

Accuracy and precision of the techniques used to measure methane emissions from ruminant animals are critically important for the success of greenhouse gas mitigation efforts. This manuscript describes the principles and operation of an automated system to monitor methane and carbon dioxide mass fluxes from the breath of ruminant animals.

自动系统（绿色饲料）的用于监视肠道的甲烷和二氧化碳从反刍动物排放

Ruminant animals (domesticated or wild) emit methane (CH4) through enteric fermentation in their digestive tract and from decomposition of manure during ...

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 ...

Experimental Study of the Relationship Between Particle Size and Methane Sorption Capacity in Shale

The amount of adsorbed shale gas is a key parameter used in shale gas resource evaluation and target area selection, and it is also an important standard for evaluating the mining value of shale gas. Currently, studies on the correlation between particle size and methane adsorption are controversial. In this study, an isothermal adsorption apparatus, the gravimetric sorption analyzer, is used to test the adsorption capacity of different particle sizes in shale to determine the relationship between the particle size and the adsorption capacity of shale. Thegravimetric method requires fewer parameters and produces better results in terms of accuracy and consistency than methods like the volumetric method. Gravimetric measurements are performed in four steps: a blank measurement, preprocessing, a buoyancy measurement, and adsorption and desorption measurements. Gravimetric measurement is presently considered to be a more scientific and accurate method of measuring the amount of adsorption; however, it is time-consuming and requires a strict measuring technique. A Magnetic Suspension Balance (MSB) is the key to verify the accuracy and consistency of this method. Our results show that adsorption capacity and particle size are correlated, but not a linear correlation, and the adsorptions in particles sieved into 40 - 60 and 60 - 80 meshes tend to be larger. We propose that the maximum adsorption corresponding to the particle size is approximately 250 &#181;m (60 mesh) in the shale gas fracturing.

We use an isothermal adsorption apparatus, the gravimetric sorption analyzer, to test the adsorption capacity of different particle sizes of shale, in order to find out the relationship between particle size and the adsorption capacity of shale.

页岩中粒径与甲烷吸附能力关系的实验研究

The amount of adsorbed shale gas is a key parameter used in shale gas resource evaluation and target area selection, and it is also an important standard ...

Measuring Stolons and Rhizomes of Turfgrasses Using a Digital Image Analysis System

Length and diameter of stolons or rhizomes are usually measured using simple rulers and calipers. This procedure is slow and laborious, so it is often used on a limited number of stolons or rhizomes. For this reason, these traits are limited in their use for morphological characterization of plants. The use of digital image analysis software technology may overcome measurement errors due to human mistakes, which tend to increase as the number and size of samples also increase. The protocol can be used for any kind of crop but is particularly suitable for forage or grasses, where plants are small and numerous. Turf samples consist of aboveground biomass and an upper soil layer to the depth of maximum rhizome development, depending on the species of interest. In studies, samples are washed from the soil, and stolons/rhizomes are cleaned by hand before analysis by digital image analysis software. The samples are further dried in a laboratory heating oven to measure dry weight; therefore, for each sample, the resultant data are total length, total dry weight, and average diameter. Scanned images can be corrected before analysis by excluding visible extraneous parts, such as remaining roots or leaves not removed with the cleaning process. Indeed, these fragments normally have much smaller diameters than stolons or rhizomes, so they can be easily excluded from analysis by fixing the minimum diameter below which objects are not considered. Stolon or rhizome density per unit area can then be calculated based on sample size. The advantage of this method is quick and efficient measurement of the length and average diameter of large sample numbers of stolons or rhizomes.

A software-based image analysis system provides an alternative method to study the morphology of stoloniferous and rhizomatous species. This protocol permits measurement of the length and diameter of stolons and rhizomes and can be applied to samples with a large amount of biomass and to a wide variety of species.

利用数字图像分析系统测量草坪的茎和根状茎

Length and diameter of stolons or rhizomes are usually measured using simple rulers and calipers. This procedure is slow and laborious, so it is often ...

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens

Many insects live above the ground as larvae and adults and as pupate below the ground. Compared to the above-ground stages of their life cycles, less attention has been paid on how environmental factors affect these insects when they pupate within the soil. The tea looper, Ectropis grisescens Warren (Lepidoptera: Geometridae), is a severe pest of tea plants and has caused huge economic losses in South China. The protocols described here aim to investigate, through multiple-choice bioassays, whether mature last-instar E. grisescens larvae can discriminate soil variables such as the substrate type and moisture content, and determine, through no-choice bioassays, the impact of the substrate type and moisture content on pupation behaviors and the emergence success of E. grisescens. The results would enhance the understanding of the pupation ecology of E. grisescens and may bring insights into soil-management tactics for suppressing E. grisescens populations. In addition, these bioassays can be modified to study the influences of various factors on the pupation behaviors and survivorship of soil-pupating pests.

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens

Here, we present a protocol to investigate the pupation preference of mature larvae of Ectropis grisescens in response to soil factors (e.g., substrate type and moisture content) using choice bioassays. We also present a protocol of no-choice bioassays to determine the factors that affect the pupation behaviors and survivorship of E. grisescens.

选择与无选择生物鉴定研究蛾 grisescens的化蛹偏好及出现的成功

Many insects live above the ground as larvae and adults and as pupate below the ground. Compared to the above-ground stages of their life cycles, less ...

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.

生态动物的长期视频跟踪: 挪威龙虾 (诺菲氏肾病) 日生物活动的案例研究

We present a protocol related to a video-tracking technique based on the background subtraction and image thresholding that makes it possible to ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

不同环境条件下生物气溶胶的组成及分布分析

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...

Automated 3D Optical Coherence Tomography to Elucidate Biofilm Morphogenesis Over Large Spatial Scales

Biofilms are a most successful microbial lifestyle and prevail in a multitude of environmental and engineered settings. Understanding biofilm morphogenesis, that is the structural diversification of biofilms during community assembly, represents a remarkable challenge across spatial and temporal scales. Here, we present an automated biofilm imaging system based on optical coherence tomography (OCT). OCT is an emerging imaging technique in biofilm research. However, the amount of data that currently can be acquired and processed hampers the statistical inference of large scale patterns in biofilm morphology. The automated OCT imaging system allows covering large spatial and extended temporal scales of biofilm growth. It combines a commercially available OCT system with a robotic positioning platform and a suite of software solutions to control the positioning of the OCT scanning probe, as well as the acquisition and processing of 3D biofilm imaging datasets. This setup allows the in situ and non-invasive automated&#160;monitoring of biofilm development and may be further developed to couple OCT imaging with macrophotography and microsensor profiling.

Microbial biofilms form complex architectures at interphases and develop into highly scale-dependent spatial patterns. Here, we introduce an experimental system (hard- and software) for the automated acquisition of 3D optical coherence tomography (OCT) datasets. This toolset allows the non-invasive and multi-scale characterization of biofilm morphogenesis in space and time.

自动 3D 光学相干断层扫描，以在大型空间尺度上阐明生物膜变形

Biofilms are a most successful microbial lifestyle and prevail in a multitude of environmental and engineered settings. Understanding biofilm ...

Leaf Area Index Estimation Using Three Distinct Methods in Pure Deciduous Stands

Accurate estimations of leaf area index (LAI), defined as half of the total leaf surface area per unit of horizontal ground surface area, are crucial for describing the vegetation structure in the fields of ecology, forestry, and agriculture. Therefore, procedures of three commercially used methods (litter traps, needle technique, and a plant canopy analyzer) for performing LAI estimation were presented step-by-step. Specific methodological approaches were compared, and their current advantages, controversies, challenges, and future perspectives were discussed in this protocol. Litter traps are usually deemed as the reference level. Both the needle technique and the plant canopy analyzer (e.g., LAI-2000) frequently underestimate LAI values in comparison with the reference. The needle technique is easy to use in deciduous stands where the litter completely decomposes each year (e.g., oak and beech stands). However, calibration based on litter traps or direct destructive methods is necessary. The plant canopy analyzer is a commonly used device for performing LAI estimation in ecology, forestry, and agriculture, but is subject to potential error due to foliage clumping and the contribution of woody elements in the field of view (FOV) of the sensor. Eliminating these potential error sources was discussed. The plant canopy analyzer is a very suitable device for performing LAI estimations at the high spatial level, observing a seasonal LAI dynamic, and for long-term monitoring of LAI.

An accurate estimation of leaf area index (LAI) is crucial for many models of material and energy fluxes within plant ecosystems and between an ecosystem and the atmospheric boundary layer. Therefore, three methods (litter traps, needle technique, and PCA) for taking precise LAI measurements were in the presented protocol.

在纯落叶支架中使用三种截然不同的方法估算叶面积指数

Accurate estimations of leaf area index (LAI), defined as half of the total leaf surface area per unit of horizontal ground surface area, are crucial for ...

Manipulation of Color Patterns in Jumping Spiders for Use in Behavioral Experiments

In the field of behavioral ecology, many experiments are designed to investigate the evolutionary purposes of colorful traits in the context of sexual selection and predation. Methods are various but mostly consist of modifying the color patterns of individuals with diverse colorants. Such techniques have been used across many vertebrate taxa, particularly in birds, but have remained underdeveloped for invertebrates because of the difficulty of effectively manipulating color in small organisms. Instead, to manipulate the appearance of invertebrates, scientists have usually modified the lighting environment to filter out certain wavelengths. However, such a method affects not only the phenotypic trait of interest but the entire appearance of the individual and its surrounding. Here, scaling down the techniques previously used on colorful birds, we present ways of manipulating the colors of small arthropods, using equally emblematic but understudied species: the colorful jumping spiders.

The goal of this protocol is to manipulate the color patterns of jumping spiders and other very small arthropods with paint in order to study questions related to sexual selection, sexual cannibalism, predation, aposematism, or any other field of animal coloration.

跳跃蜘蛛颜色模式的处理及其在行为实验中的应用

In the field of behavioral ecology, many experiments are designed to investigate the evolutionary purposes of colorful traits in the context of sexual ...