笔者想感谢所有的新英格兰商业lobstermen的船，我们被允许从品尝 - 他们的合作是非常赞赏。此外，我们要感谢小龙虾湾公司（纽因顿，NH）和Champlin的的海鲜（点朱迪思，RI）谁允许我们在他们的设施进行采样。下面UNH学生在本研究提供了宝贵的援助：海利白色，弗朗索瓦·莫里森，莎拉Havener，奥德拉Chaput的，和五月格罗斯。该项目由赠款从新罕布什尔州SeaGrant到WHW（项目＃R/CFR-11）和UNH海洋项目资助，以联合研究小组的支持。

A部分：现场抽样技术 1。女性三围<ol><li>对于每个雌性龙虾，测量头胸甲长（CL）和所述第二腹节到使用一对卡尺的最接近1.0毫米的宽度。 注意：第二腹节进行测量，因为腹部（其中女性随身携带的蛋）的宽度是性成熟的一个指标。此外，它可能是有利的，以确定蜕皮阶段以后配合生殖阶段这些数据（参见步骤1.2）。 </li><li>夹一小部分远端关闭腹腔腹足之一（龙虾标准的方法论协议）和腹足清洁海水储存后用解剖显微镜观察9。 注意：此步骤可能也露出水泥腺体对腹肢，这是女性正准备挤出它们的卵10指示的存在。 </li></ol> 2。樱雪针rtion <ol><li>将注射器针头（18克， 见表1）〜1厘米的精液接受器，向着尾巴〜45°（ 图1）的角度。 注意：在龙虾与完整的精子塞，针可能会遇到一些阻力插入过程中，因为它是通过精子插头之前将精子的浓度最高的精液容器的底部（ 图1）推压。 注意：插头的存在，本身就是一个可靠的指标，有龙虾交配。在一些动物可能有很小的阻力，这可能是因为没有精囊存在，精囊是小的，它没有硬化成一个插头还，或者它可能已经被用于受精的鸡蛋中的一个或多个离合器。 </li></ol><img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig1highres.jpg" src="/files/ftp_upload/50498/50498fig1.jpg" widTH =“600px的”/&gt; 图1。针插入和视图的龙虾精液插座。插入钝头针头插入龙虾的精液接受器中提取精子样本。针的角度和深度都在取得一致的样品是至关重要的。右：查看已解剖和垂直分割的一个开创性的插座。精子塞占据了容器的顶部或最外部分，而精子位于所述容器的底部（最深点）。需要注意的是低于这个深度取样可能刺破插座，并导致血液污染的样品中是很重要的。 <a href="https://www.jove.com/files/ftp_upload/50498/50498fig1highres.jpg" target="_blank">点击这里查看大图</a> 。 3。穿透精子插件<ol><li>最初的插入后，陡针头至接近垂直的角度（垂直于身体轴），并非常缓慢地开始工作，它向下插入插座。 </li><li>保持倾斜一个角度针，使针尖朝向尾部略尖。在这种情况下，针尖压缩所述插头的顶部，直到它最终渗透到基质中。 </li><li>继续针头慢慢往下压，直到感觉到阻力再一次 - 这是插座的底部，并在那里被抽取的样品。 </li></ol> 4。去除精子<ol><li>从容器的底部取出的样品（通常固体插塞材料，液体精子和其它流体）和采样存入一个标记2.0毫升塑料管。 </li><li>冲洗容器用〜0.1-0.5毫升冷海水。 </li><li>在同一管除去约0.3ml的流体和地点。 </li><li>冰商店精子样本，直到他们可以在实验室进行检查。 B部分：精子样本在实验室考试 注：从个人的升精液的容器取出样品<html> obsters进行检查，以确定是否精子细胞存在或不存在。 </li><li>从每个塑料管，地方与盖玻片载玻片上，并认为在100X有一个复式显微镜取出〜50微升的液体。 </li><li>分数的样本为：精子稀少，众多的，或不存在（此研究没有实际的量化精子数， 图2）。 </li></ol><img alt="图2" fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig2highres.jpg" src="/files/ftp_upload/50498/50498fig2.jpg" width="600px" /> 图2。龙虾的精子样本图像。从女性的精液接受器取，作为观察与复合式显微镜放大40倍龙虾精子样本图像。图像上的左显示，含有大量的精子，而另一（右）示出了一个稀疏样本（比例尺= 10μm）的一个样本。jpg“格式目标=”_blank“&gt;点击这里查看大图。

<ol>
	<li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> FAO and Agriculture Organization of the United Nations). Fisheries and Aquaculture Department. , (2012).</li><li>MacDiarmid, A., Butler, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sperm+economy+and+limitation+in+spiny+lobsters.">Sperm economy and limitation in spiny lobsters.</a> Behav. Ecol. Sociobiol. 46, 14-24 (1999).</li><li>Kendall, M. S., Wolcott, D. L., Wolcott, T. G., Hines, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+male+size+and+mating+history+on+sperm+content+of+ejaculates+of+the+blue+crab+Callinectes+sapidus.">Influence of male size and mating history on sperm content of ejaculates of the blue crab Callinectes sapidus.</a> 230, 235-240 (2002).</li><li>Sato, T., Ashidate, M., Goshima, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+male+mating+frequency+and+male+size+on+ejaculate+size+and+reproductive+success+of+female+spiny+king+crab,+Parolithodes+brevipes.">Effects of male mating frequency and male size on ejaculate size and reproductive success of female spiny king crab, Parolithodes brevipes.</a> Mar. Ecol. Prog. Ser. 296, 251-262 (2005).</li><li>Sato, T., Goshima, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impacts+of+male-only+fishing+and+sperm+limitation+in+manipulated+populations+of+an+unfished+crab,+Hapalogaster.">Impacts of male-only fishing and sperm limitation in manipulated populations of an unfished crab, Hapalogaster.</a> 313, 193-204 (2006).</li><li>Rondeau, A., Sainte-Marie, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variable+mate-guarding+time+and+sperm+allocation+by+male+snow+crabs+(Chionoecetes+opilio)+in+response+to+sexual+competition,+and+their+impact+on+the+mating+success+of+females.">Variable mate-guarding time and sperm allocation by male snow crabs (Chionoecetes opilio) in response to sexual competition, and their impact on the mating success of females.</a> Biol. Bull. 201, 204-217 (2001).</li><li>Waddy, S. L., Aiken, D. E., Bauer, R. T., Martin, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Mating and insemination in the Ameri can lobster,Homarus americanus. Crustacean sexual behavior. , 126-144 (1990).</li><li>Johnson, K. J., Goldstein, J. S., Watson, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+methods+for+determining+the+fertility+status+in+early-stage+American+lobster,+Homarus+americanus,+eggs.">Two methods for determining the fertility status in early-stage American lobster, Homarus americanus, eggs.</a> J. Crust. Biol. 31, 693-700 (2011).</li><li>Aiken, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proecdysis,+setal+development,+and+molt+prediction+in+the+American+lobster+(Homarus).">Proecdysis, setal development, and molt prediction in the American lobster (Homarus).</a> 30, 1337-1344 (1973).</li><li>Aiken, D. E., Waddy, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cement+gland+development,+ovary+maturation,+and+reproductive+cycles+in+the+American+lobster+Homarus+americanus.">Cement gland development, ovary maturation, and reproductive cycles in the American lobster Homarus americanus.</a> J. Crust. Biol. 2, 315-327 (1982).</li><li>Pugh, T. L., Goldstein, J. S., Lavalli, K. L., Clancy, M., Watson, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=At-sea+determination+of+female+American+lobsters+(Homarus+americanus)+mating+activity:+Patterns+vs.+expectations.">At-sea determination of female American lobsters (Homarus americanus) mating activity: Patterns vs. expectations.</a> Fish. Res. 147, 327-337 (2013).</li><li>Bauer, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phylogenetic+trends+in+sperm+transfer+and+storage+complexity+in+decapod+crustaceans.">Phylogenetic trends in sperm transfer and storage complexity in decapod crustaceans.</a> J. Crust. Biol. 6, 313-325 (1986).</li><li>Talbot, P., Helluy, S., Factor, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproduction+and+embryonic+development.">Reproduction and embryonic development.</a> Biology of the Lobster: Homarus americanus. , 177-216 (1995).</li><li>Cooper, R. A., Uzmann, J. R., Cobb, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ecology of juvenile and adult Homarus. The biology and management of lobsters. 2, 97-142 (1980).</li><li>Estrella, B. T., McKiernan, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catch-per-unit-effort+and+biological+parameters+from+the+Massachusetts+coastal+lobster+(Homarus+americanus)+resource:+description+and+trends.">Catch-per-unit-effort and biological parameters from the Massachusetts coastal lobster (Homarus americanus) resource: description and trends.</a> NOAA Tech. Rep. NMFS. 81, (1989).</li><li>Waddy, S. L., Aiken, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+fertilization+and+consecutive+spawning+in+large+American+lobster,+Homarus+americanus.">Multiple fertilization and consecutive spawning in large American lobster, Homarus americanus.</a> Can. J. Fish. Aquat. Sci. 43, 2291-2294 (1986).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atlantic+States+Marine+Fisheries+Commission+(ASMFC).+American+lobster+stock+assessment+report+for+peer+review.">Atlantic States Marine Fisheries Commission (ASMFC). American lobster stock assessment report for peer review.</a> ASMFC Stock Assessment. , (2009).</li></ol>

作者宣称没有利益冲突。

这部影片和纸提纲，并展示用于确定雌性龙虾已经成功交配的方法。这种方法是很简单的，它可以由多种用户用于龙虾海上大规模抽样。该方法是基于检测精荚的传递从男女龙虾交配时精液容器的存在。这些精荚局部变硬成精子塞它们存放在容器后不久，因此，精子塞的存在也表明交配成功。在这项研究中，以及一个随后的更大的研究，已确定，大部分精子插头已精子与它们相关联11。然...

<html>美国龙虾（ 美洲螯龙虾 ）是在北大西洋（〜56,000吨，2011年，价值超过390,000,000元）1最有生产力的渔场之一。然而，有关于这个物种的野生种群的繁殖动力学普遍缺乏了解。生成繁殖输出更准确的估计，包括个人的积极参与繁殖的数量和大小，可以提高该股的评估过程。例如，女性是从实现其全部潜能的生殖防止因精子限制已被确定为几个商业开发海洋甲壳类动物，包括一个担忧：多刺龙虾2，蓝蟹3，帝王蟹4，石蟹5，和雪蟹6。总的目标是，以确定是否精子的限制可能是在美国龙虾渔业某些地区的一个因素也是如此。 NT“&gt;，同时进行与产卵雌龙虾无关的标记研究（有明显完好无损，最近挤压蛋）的龙虾潜在的精子限制问题的第一个征象进行观察。夺回渔民报告表明，〜这些动物的15％已下降他们的鸡蛋离合器只有1-2个月后的工作假设是，一些龙虾携带了那些尚未被受精卵，并作为结果，“掉下来”7。从随后的研究的初步数据已经证实的事实，将龙虾挤出鸡蛋即使他们没有成功交配而这些未受精的卵子只携带〜1 第八个月，因此，给出的观测女性携带未受精的卵子在自然种群中，我们试图确定到精子的限制可能会造成程度由性成熟龙虾再现的次最大水平。为了实现这一目标，一种技术被开发，以检测spermato交配过程中phore沉积男性。 本文和视频描述为确定女性龙虾交配成功开发出一种无创，简便的方法。该技术可以快速，可靠地使用在海上，无论是乘坐研究或商业渔船。这种抽样方法的细节，以及一些有代表性的研究结果说明了该技术的应用。

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a noninvasive method for in situ determination of mating success in female american lobsters (homarus americanus)

尽管是在西北大西洋最有生产力的渔场之一，还有许多未知的关于美国龙虾自然繁殖动态。最近在利用甲壳类人群（螃蟹和龙虾）的研究表明，在有些情况下成熟的女性是无法充分实现其生殖潜力，由于精子的限制。为了检验这种可能性在美国龙虾渔业的不同区域，一个可靠的无创方法，在海上采样大批女性龙虾的开发。该方法涉及将一个钝头针头插入女性的精液接受器来确定精子塞的存在或不存在和撤回，可检查精子的存在的样品。一系列控制研究是在码头和在实验室中进行的，以测试该技术的可靠性。这些努力entailed抽样294女龙虾，确认存在Õ发精子插头是精液的容器内的可靠指标，因此，交配。本文详细说明的方法及从采样的总位女性的子集​​所获得的结果。从乔治的银行和海角安，马取样的230女龙虾（尺寸范围= 71-145毫米，背甲长），90.3％呈阳性的精子。潜在的解释为没有精子在女性的一些包括：不成熟（缺乏生理成熟），精子插件被用来卵子受精的离合器故障后，和缺乏交配活动。调查表明，该技术用于检查雌性龙虾的交配成功是一个可靠的代理，可用于在现场对文件生殖活动中自然种群。

<html>这里报道的数据是针对源于近岸和近海地区的女性龙虾的一个子集。近岸组包括被抓获的近海角安，马萨诸塞州沿海水域及运送冷却器实验室检查44龙虾。共有186近海龙虾被抓获乔治银行和采样在新罕布什尔州举行的商业设施。 精子样品是从所有的动物中，根据上述方法获得的，并立即检查精子的存在与否。总共在这个子集的所有女性90.3％有精子在他们的精液的容器。?...

Watch this Scientific Journal Video about A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus at JoVE.com

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

渔业引起的变化，以利用甲壳类水产，如美国龙虾捕捞业，有可能影响他们的生育动态，导致交配成功的减少。这项研究的目标是开发以确定女性龙虾，可能是生理上或功能上的成熟的交配成功一种无创性的方法。

一种非侵入性的方法<em&gt;原位</em&gt;交配成功的女美国龙虾​​（测定<em&gt;美洲螯龙虾</em&gt;）

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American ...

a-noninvasive-method-for-situ-determination-mating-success-female

Research

JoVE Journal

Environment

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

10.9K 次观看.  University of New Hampshire. 渔业引起的变化，以利用甲壳类水产，如美国龙虾捕捞业，有可能影响他们的生育动态，导致交配成功的减少。这项研究的目标是开发以确定女性龙虾，可能是生理上或功能上的成熟的交配成功一种无创性的方法。 

视频: A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

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

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

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

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

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

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

科研

环境科学

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

VacuSIP，对改进的方法INEX<em&gt;原位</em&gt;颗粒物与测量溶解的化合物加工后主动悬挂给料机

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

Optimized Procedure for Determining the Adsorption of Phosphonates onto Granular Ferric Hydroxide using a Miniaturized Phosphorus Determination Method

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide (GFH), with little effort and high reliability. The phosphonate, e.g., nitrilotrimethylphosphonic acid (NTMP), is brought into contact with the GFH in a rotator in a solution buffered by an organic acid (e.g., acetic acid) or Good buffer (e.g., 2-(N-morpholino)ethanesulfonic acid) [MES] and N-cyclohexyl-2-hydroxyl-3-aminopropanesulfonic acid [CAPSO]) in a concentration of 10 mM for a specific time in 50 mL centrifuge tubes. Subsequently, after membrane filtration (0.45 &#181;m pore size), the total phosphorus (total P) concentration is measured using a specifically developed determination method (ISOmini). This method is a modification and simplification of the ISO 6878 method: a 4 mL sample is mixed with H2SO4 and K2S2O8 in a screw cap vial, heated to 148-150 &#176;C for 1 h and then mixed with NaOH, ascorbic acid and acidified molybdate with antimony(III) (final volume of 10 mL) to produce a blue complex. The color intensity, which is linearly proportional to the phosphorus concentration, is measured spectrophotometrically (880 nm). It is demonstrated that the buffer concentration used has no significant effect on the adsorption of phosphonate between pH 4 and 12. The buffers, therefore, do not compete with the phosphonate for adsorption sites. Furthermore, the relatively high concentration of the buffer requires a higher dosage concentration of oxidizing agent (K2S2O8) for digestion than that specified in ISO 6878, which, together with the NaOH dosage, is matched to each buffer. Despite the simplification, the ISOmini method does not lose any of its accuracy compared to the standardized method.

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide, with little effort and high reliability. In a buffered solution, the phosphonate is brought into contact with the adsorbent using a rotator and then analyzed via a miniaturized phosphorus determination method.

用小型化磷测定法测定膦酸在粒状氢氧化铁上吸附的优化方法

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric ...

In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples

Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ocean temperatures and ocean acidification, corals are increasingly under threat. To tackle these challenges, advances in cell and molecular biology have proven to be crucial for diagnosing the health of corals. Modifying some of the techniques commonly used in human medicine could greatly improve researchers&#39; ability to treat and save corals. To address this, a protocol for in situ hybridization used primarily in human medicine and evolutionary developmental biology has been adapted for use in adult corals under stress.
The purpose of this method is to visualize the spatial expression of an RNA probe in adult coral tissue that has been embedded in paraffin and sectioned onto glass slides. This method focuses on removal of the paraffin and rehydration of the sample, pretreatment of the sample to ensure permeability of the sample, pre-hybridization incubation, hybridization of the RNA probe, and visualization of the RNA probe. This is a powerful method when using non-model organisms to discover where specific genes are expressed, and the protocol can be easily adapted for other non-model organisms. However, the method is limited in that it is primarily qualitative, because expression intensity can vary depending on the amount of time used during the visualization step and the concentration of the probe. Furthermore, patience is required, as this protocol can take up to 5 days (and in many cases, longer) depending on the probe being used. Finally, non-specific background staining is common, but this limitation can be overcome.

In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples

The goal of this protocol is to perform in situ hybridization on adult coral samples that have been embedded in paraffin and sectioned onto glass slides. This is a qualitative method used to visualize the spatial expression of an RNA anti-sense probe in paraffin-embedded tissues.

就地石蜡镶嵌成人珊瑚样品的杂交技术

Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ...

A Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Identification of Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many countries around the world. Severe yield losses are caused by direct feeding, and even more importantly, also by the transmission of more than 100 harmful plant pathogenic viruses. As for other invasive pests, increased international trade facilitates the dispersal of B. tabaci to areas beyond its native range. Inspections of plant import products at points of entry such as seaports and airports are, therefore, seen as an important prevention measure. However, this last line of defense against pest invasions is only effective if rapid identification methods for suspicious insect specimens are readily available. Because the morphological differentiation between the regulated B. tabaci and close relatives without quarantine status is difficult for non-taxonomists, a rapid molecular identification assay based on the loop-mediated isothermal amplification (LAMP) technology has been developed. This publication reports the detailed protocol of the novel assay describing rapid DNA extraction, set-up of the LAMP reaction, as well as interpretation of its read-out, which allows identifying B. tabaci specimens within one hour. Compared to existing protocols for the detection of specific B. tabaci biotypes, the developed method targets the whole B. tabaci species complex in one assay. Moreover the assay is designed to be applied on-site by plant health inspectors with minimal laboratory training directly at points of entry. Thorough validation performed under laboratory and on-site conditions demonstrates that the reported LAMP assay is a rapid and reliable identification tool, improving the management of B. tabaci.

A Loop-mediated Isothermal Amplification LAMP Assay for Rapid Identification of Bemisia tabaci

This paper reports the protocol for a rapid identification assay for Bemisia tabaci based on loop-mediated isothermal amplification (LAMP) technology. The protocol requires minimal laboratory training and can, therefore, be implemented on-site at points of entry for plant imports such as seaports and airports.

循环介导等温扩增 (灯管) 法快速识别烟粉虱

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many ...

Implantation of a New Micro Acoustic Tag in Juvenile Pacific Lamprey and American Eel

Juvenile Pacific Lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology has been identified as a way to obtain more detailed information on movement and behavior across a broader spatial scale than is possible with other known technology. The purpose of this method is to provide a detailed step by step instruction on tag implantation for both lampreys and eel. For laboratory studies using actively migrating juvenile Pacific Lamprey (120-160 mm), we determined that the presence of the tag did not alter the swimming ability between tagged and untagged Individuals or have any significant tag loss (&lt;3%). Similar results were determined during laboratory testing of Yellow phase American Eels (113-175 mm). No mortality occurred during a 38-day holding period and there was minimal tag loss (3.8%). The presence of the tag did not have any significant effect on the swimming ability or survival of tagged eels compared to untagged controls and there was minimal tag loss.

This article illustrates the procedure for implanting a micro acoustic tag in juvenile Pacific Lamprey and American Eel. Our application is based on laboratory trials, which indicate that acoustic tags can be successfully implanted with no apparent effects on swimming ability or survival and minimal tag loss.

一种新的微声学标记在少年太平洋兰普瑞和美洲鳗鱼中的植入

Juvenile Pacific Lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology ...

A Telemetric, Gravimetric Platform for Real-Time Physiological Phenotyping of Plant&ndash;Environment Interactions

Food security for the growing global population is a major concern. The data provided by genomic tools far exceeds the supply of phenotypic data, creating a knowledge gap. To meet the challenge of improving crops to feed the growing global population, this gap must be bridged.
Physiological traits are considered key functional traits in the context of responsiveness or sensitivity to environmental conditions. Many recently introduced high-throughput (HTP) phenotyping techniques are based on remote sensing or imaging and are capable of directly measuring morphological traits, but measure physiological parameters mainly indirectly.
This paper describes a method for direct physiological phenotyping that has several advantages for the functional phenotyping of plant&#8211;environment interactions. It helps users overcome the many challenges encountered in the use of load-cell gravimetric systems and pot experiments. The suggested techniques will enable users to distinguish between soil weight, plant weight and soil water content, providing a method for the continuous and simultaneous measurement of dynamic soil, plant and atmosphere conditions, alongside the measurement of key physiological traits. This method allows researchers to closely mimic field stress scenarios while taking into consideration the environment&#8217;s effects on the plants&#8217; physiology. This method also minimizes pot effects, which are one of the major problems in pre-field phenotyping. It includes a feed-back fertigation system that enables a truly randomized experimental design at a field-like plant density. This system detects the soil-water-content limiting threshold (&#952;) and allows for the translation of data into knowledge through the use of a real-time analytic tool and an online statistical resource. This method for the rapid and direct measurement of the physiological responses of multiple plants to a dynamic environment has great potential for use in screening for beneficial traits associated with responses to abiotic stress, in the context of pre-field breeding and crop improvement.

A Telemetric, Gravimetric Platform for Real-Time Physiological Phenotyping of Plant&amp;#8211;Environment Interactions

This high-throughput, telemetric, whole-plant water relations gravimetric phenotyping method enables direct and simultaneous real-time measurements, as well as the analysis of multiple yield-related physiological traits involved in dynamic plant&#8211;environment interactions.

植物与环境相互作用实时生理表型的遥测重力平台

Food security for the growing global population is a major concern. The data provided by genomic tools far exceeds the supply of phenotypic data, creating ...

Mistletoe Eradicator - A Novel Tool for Simultaneous Mechanical and Chemical Control of Mistletoe

Some species of mistletoes, an aerial plant hemiparasites, play a Janus-like role in ecosystems by being rich biodiversity hotspots due to their trophic and topic associations with various organisms and at the same time acting as biological pirates, exhausting the host trees by stealing water and nutrients via the haustorium. Many crop and timber trees are attacked by mistletoes, for example, dwarf mistletoes parasitize conifers, representatives of genus Tapinanthus target guava, and Phoradendron mistletoe target pecans. Recently, an increasing infestation of the European mistletoe (Viscum album subsp. album L.) in monoculture stands of common walnuts (Juglans regia L.)&#160;was recorded for the first time during 2018 in different regions of Kashmir Valley, India. The common chemical approach based on the spraying of mistletoe exophytes with the growth regulators, ethephon and glyphosate, was not efficient since V. album resprouted in the next vegetation season. Hand pruning of mistletoe was time-consuming, and a polyethylene covering resulted in the localized rotting of host tissues. The most efficient approach to eradicate mistletoe was the use of a novel tool called Mistletoe Eradicator, which allows to use mechanical and chemical means of mistletoe control simultaneously. It consists of a manipulating pole with the terminal pruning saw having a container filled with the growth regulator, from which the liquid slowly drops onto the saw blade through the nozzle. It allows working both from the ground and in the tree canopy. After the targeted application of 5%-10% ethephon or glyphosate to the base of the just pruned V. album stems (stubs), exophyte resprouting was significantly reduced 9 months after the pruning as compared to the only pruned or sprayed exophytes. An alternative variant of Mistletoe Eradicator was constructed in Ukraine filled just with a food color additive to track the liquid dropping and was successfully tested.

The protocol describes an easy-to-handle tool called Mistletoe Eradicator which combines mechanical and chemical approaches and is efficient in the control of mistletoe infestation on the host trees. The manuscript uses the European mistletoe (Viscum album subsp. album L.) as an example.

槲寄生根除器 - 同时机械和化学控制槲寄生的新型工具

Some species of mistletoes, an aerial plant hemiparasites, play a Janus-like role in ecosystems by being rich biodiversity hotspots due to their trophic ...

A Complete Method for Evaluating the Performance of Photocatalysts for the Degradation of Antibiotics in Environmental Remediation

Various antibiotics such as tetracycline, aureomycin, amoxicillin, and levofloxacin are found in large quantities in groundwater and soil systems, potentially leading to the development of resistant and multi-drug resistant bacteria, posing a threat to humans, animals, and environmental systems. Photocatalytic technology has attracted keen interest due to its rapid and stable treatment and direct use of solar energy. However, most studies evaluating the performance of semiconductor catalysts for the photocatalytic degradation of organic pollutants in water are currently incomplete. In this paper, a complete experimental protocol is designed to comprehensively evaluate the photocatalytic performance of semiconductor catalysts. Herein, rhombic dodecahedral silver phosphate was prepared by a simple solvent phase synthesis method at room temperature and atmospheric pressure. BrSubphthalocyanine/Ag3PO4 heterojunction materials were prepared by the solvothermal method. The catalytic performance of as-prepared materials for the degradation of tetracycline was evaluated by studying different influencing factors such as catalyst dosage, temperature, pH, and anions at atmospheric pressure using a 300 W xenon lamp as a simulated solar light source and a light intensity of 350 mW/cm2. Compared with the first cycle, the constructed BrSubphthalocyanine/Ag3PO4 maintained 82.0% of the original photocatalytic activity after five photocatalytic cycles, while the pristine Ag3PO4 maintained only 28.6%. The stability of silver phosphate samples was further tested by a five-cycle experiment. This paper provides a complete process for evaluating the catalytic performance of semiconductor catalysts in the laboratory for the development of semiconductor catalysts with potential for practical applications.

Presented here is a protocol to explore a universal set of experimental procedures for comprehensive laboratory evaluation of photocatalysts in the field of environmental purification, using the example of photocatalytic removal of antibiotic organic pollutant molecules from water by phthalocyanine sensitized silver phosphate composites.

一种评价光催化剂降解环境修复中抗生素性能的完整方法

Various antibiotics such as tetracycline, aureomycin, amoxicillin, and levofloxacin are found in large quantities in groundwater and soil systems, ...

In Situ Soil Moisture Sensors in Undisturbed Soils

Soil moisture directly affects operational hydrology, food security, ecosystem services, and the climate system. However, the adoption of soil moisture data has been slow due to inconsistent data collection, poor standardization, and typically short record duration. Soil moisture, or quantitatively volumetric soil water content (SWC), is measured using buried, in situ sensors that infer SWC from an electromagnetic response. This signal can vary considerably with local site conditions such as clay content and mineralogy, soil salinity or bulk electrical conductivity, and soil temperature; each of these can have varying impacts depending on the sensor technology.
Furthermore, poor soil contact and sensor degradation can affect the quality of these readings over time. Unlike more traditional environmental sensors, there are no accepted standards, maintenance practices, or quality controls for SWC data. As such, SWC is a challenging measurement for many environmental monitoring networks to implement. Here, we attempt to establish a community-based standard of practice for in situ SWC sensors so that future research and applications have consistent guidance on site selection, sensor installation, data interpretation, and long-term maintenance of monitoring stations.
The videography focuses on a multi-agency consensus of best-practices and recommendations for the installation of in situ SWC sensors. This paper presents an overview of this protocol along with the various steps essential for high-quality and long-term SWC data collection. This protocol will be of use to scientists and engineers hoping to deploy a single station or an entire network.

In Situ Soil Moisture Sensors in Undisturbed Soils

The determination of soil water content is a critical mission requirement for many state and federal agencies. This protocol synthesizes multi-agency efforts to measure soil water content using buried in situ sensors.

原位 未受干扰土壤中的土壤湿度传感器

Soil moisture directly affects operational hydrology, food security, ecosystem services, and the climate system. However, the adoption of soil moisture ...