彼らの協力ははるかに高く評価されている - 著者は、そのボート我々はからサンプルを許可されたすべてのニューイングランドの商業lobstermanの複数形に感謝したいと思います。加えて、我々はリトルベイロブスターカンパニー（ニューイントン、ニューハンプシャー州）、私たちは彼らの施設でサンプリングを行うことが許さチャンプリンのシーフード（ポイントジュディス、RI）を感謝します。次のUNHの学生がこの研究を通して非常に貴重な支援を提供：ヘイリーホワイト、フランソワーズ·モリソン、サラHavener、オードラシャプー法、5月グロースを。このプロジェクトは、WHW（プロジェクト＃のR/CFR-11）と共同研究会へのUNHマリンプログラム付与するには、NHのSeaGrantからの補助金によって支えられている。

パートA：フィールドサンプリングテクニック 1。女性の測定<ol><li>各女性のロブスターは、甲長（CL）およびキャリパーのペアを使用して、最も近い1.0ミリメートルの第二の腹部のセグメントの幅を測定。 注：（雌は卵を運ぶ）腹部の幅は性成熟の指標であるので、第2腹節が測定される。また、（ステップ1.2参照）に保存生殖成長段階で、これらのデータと一致するように脱皮段階を確認するために有利であり得る。 </li><li>腹部の腹肢（ロブスター内の標準方法論のプロトコル）の1から小さな遠位部をクリップし、後に解剖顕微鏡9で観察するためのクリーンな海水で腹肢を保存する。 注：このステップはまた、女性が自分の卵10を押し出すために準備をしている指標である腹肢、上のセメント腺の存在を明らかに可能性があります。 </li></ol> 2。針INSErtion <ol><li> （、18 G、表1を参照）注射針を挿入〜45°（ 図1）〜尾に向かって角度のついた精液のレセプタクルに1cmの。 注：それは前に精液レセプタクル（ 図1）の下部にある精子の最高濃度に到達する精子のプラグを介してプッシュされるように完全な精子プラグが付いているロブスターでは、針は、挿入の間に多少の抵抗を満たすことがあります。 注：プラグの存在は、それ自体では、ロブスターが交尾している信頼性の高い指標である。いくつかの動物ではない精包が存在しない可能性があるため、ほとんど抵抗があってもよい、精包は、それがまだプラグに硬化していないか、またはそれが既に卵の一つ以上のクラッチを受精するのに使用されている可能性が小さい。 </li></ol><img alt="図1" fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig1highres.jpg" src="/files/ftp_upload/50498/50498fig1.jpg" wid目= &quot;600PX&quot; /&gt; 図1。針の挿入やロブスター精液レセプタクルの眺め。精子サンプルを抽出するロブスターの精液レセプタクルに鈍先端が針を挿入。針の角度と深さは、両方の一貫性のあるサンプルを得るために重要である。右：解剖し、縦方向に分割されている精液のレセプタクルの眺め。精子が容器の下部（最深部）に位置している間、精子のプラグは、コンセントの上部または最も外側の部分を占めている。それは、この深さ以下のサンプリングはレセプタクルを穿刺し、血液で汚染されたサンプルをもたらすかもしれないことに注意することが重要である。 <a href="https://www.jove.com/files/ftp_upload/50498/50498fig1highres.jpg" target="_blank">拡大画像を表示するには、ここをクリックしてください</a> 。 3。精子プラグを貫通<ol><li>最初の挿入後に、（体軸に垂直な）ほぼ垂直に、針の角度を急勾配にし、非常にゆっくりと作業を開始下向きレセプタクルに。 </li><li>先端が尾の方に少し指摘されるような角度で傾いた針を保管してください。それが最終的にマトリックスに浸透するまで、この場合、針の先端は、プラグの上部を圧縮している。 </li><li>抵抗をもう一度感じられるまで徐々に下方向に針をプッシュし続ける - これは、容器の底部にあり、どこでサンプルが採取されている。 </li></ol> 4。精子の取り外し<ol><li>レセプタクルの底からのサンプル（しばしば固体プラグ材料、液状精子と他の流体）を外し、ラベルされた2.0ミリリットルのプラスチックチューブにサンプルを堆積させる。 </li><li>冷たい海水〜0.1〜0.5ミリリットルとレセプタクルをフラッシュします。 </li><li>同じチューブ内の流体と場所の約0.3ミリリットルを削除します。 </li><li>氷上ストア精子サンプル彼らは実験室で検査することができるようになるまで。 パートB：ラボでの精子サンプルの検討 注：個々のLの精液コンセントから取り外しサンプル<html> obstersは精子細胞が存在するか存在しないかどうかを決定するために検討すべきである。 </li><li>各プラスチック製のチューブ、カバーガラスでガラススライド上の場所、および複合顕微鏡で100倍の視野から〜流体の50μLを削除します。 </li><li>としてサンプルスコア：希少数多く、または存在しない精子（本研究では、実際の精子数は 、 図2を定量化しなかった）。 </li></ol><img alt="図2" fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig2highres.jpg" src="/files/ftp_upload/50498/50498fig2.jpg" width="600px" /> 図2。ロブスター精子サンプル画像。40X倍率の複合顕微鏡で見たときに、女性の精液コンセントから取ったロブスター精子サンプルの画像。左の画像は、他の（右）スパースサンプル（スケールバー= 10μm）を示しているが、多数の精子が含まれていたサンプルを示しています。。JPG「標的= &quot;_blank&quot;&gt;拡大画像を表示するにはここをクリックしてください。

<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. 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著者らは、利害の衝突を宣言していません。

この動画と紙輪郭と雌ロブスターが正常交配しているかどうかを決定するための方法を実証する。このアプローチは、ユーザーの様々な海でのロブスターの大規模なサンプリングのために使用できることが十分に単純である。この方法は、交配の際にオスからメスのロブスターに渡される精液の容器に精包の存在を検出することに基づいています。これら精包は、部分的に、それらが容器内?...

<html>アメリカンロブスター（ ロブスターのamericanus）は、北大西洋（〜3.9億ドルかけて評価され、2011年に56,000 MT、）1の中で最も生産的な漁業の一つです。しかし、野生の個体群におけるこの種の生殖ダイナミクスに関する理解の一般的な不足があります。積極的に再生に参加する人々の数とサイズを含む生殖出力のより正確な推定値を、生成することは、資源評価プロセスを改善することがあります。イセエビ2、ワタリガニ3、キングカニ4、石のカニ5、およびズワイガニたとえばによる精子の制限のため、完全な生殖能力を達成することを防止することができる女性はを含むいくつかの商業的搾取の海洋甲殻類の懸念として同定されている6。全体的な目標は、精子の制限が同様にアメリカのロブスター漁業のある領域に要因になるかどうかを判断することです。 NT &quot;&gt;（目に見えて無傷で、最近になって押し出さ卵）抱卵雌のロブスターと関係のないタグ付けの研究を行いながらロブスターの潜在的な精子の制限の問題の最初の兆候が観察された。〜これらの動物の15％が低下したことを示した漁師からレポートを奪還彼らの卵のクラッチわずか1〜2ヶ月後。作業仮説は、いくつかのロブスターは、結果として、7 &#39;落ちた&#39;、受精とされていなかった。その後の研究の予備的データはロブスターがあるという事実を確認した卵を運んでいたということでした彼らが成功して交配しておらず、これらの未受精卵にのみ〜1ヶ月8用に運ばれていても、卵を押し出しているため、自然集団における未受精卵を運ぶ人の女性の観察を考えると、我々は精子の制限はに貢献するかもしれない程度を決定しようとした性的に成熟したロブスターによる再生の最大下のレベル。この目標を達成するために、技術がspermatoを検出するために開発されました交配中に男性が堆積phore。 本論文では、ビデオは、女性のロブスターの交尾の成功を確認するために開発された非侵襲的な、簡単な方法を説明します。技術は海で、いずれかの研究や商業漁船に乗って迅速かつ確実に利用することができる。このサンプリング方法の詳細だけでなく、いくつかの代表的な知見は、技術の適用を例示するために提示される。

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			<td height="40" style="height:40px;width:163px;">Calipers: Mitutoyo Digimatic</td>
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			<td height="40" style="height:40px;width:163px;">3 mL Luer-Lok&#8482; syringe</td>
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			<td height="60" style="height:60px;width:163px;">Monoject aluminum hub blunt needles, 18-G x 1</td>
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			<td height="40" style="height:40px;width:163px;">2.0 mL plastic storage tubes</td>
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			<td height="40" style="height:40px;width:163px;">Styrofoam cooler with ice</td>
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			<td height="40" style="height:40px;width:163px;">Compound light microscope</td>
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			<td height="41" style="height:41px;width:163px;">Pipettor with disposable tips</td>
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a noninvasive method for in situ determination of mating success in female american lobsters (homarus americanus)

北西大西洋で最も生産漁業の1であるにもかかわらず、多くのアメリカのロブスターの自然な生殖ダイナミクスについて不明である。悪用甲殻類の個体数（カニやエビ）での最近の研究は、成熟したメスによる精子の制限により、完全な生殖能力を達成することができない事情があることを示唆している。アメリカンロブスター漁業の異なる領域にこの可能性を検討するために、信頼性があり、非侵襲的方法は、海上で女性のロブスターを大量にサンプリングするために開発されました。この方法は、精子プラグの有無を判定し、精子の存在について調べることができる試料を撤回する雌の精液容器に鈍い先端が針を挿入することを含む。制御一連の研究は、この技術の信頼性をテストするために、ドックで、実験室で行った。こうした努力は、294の女性ロブスターを確認するために、サンプリングを伴うことプレゼンスOFA精子プラグは、信頼性の高い容器の中の精子の指標、したがって、交配した。本論文では、方法論とサンプリングされた合計の女性のサブセットから得られた結果の詳細を示します。ジョージの銀行とCape Annのマサチューセッツ（甲長サイズ範囲= 71〜145ミリメートル）からサンプリングされた230の女性ロブスターのうち、90.3％が精子陽性であった。一部の女性では精子の不在のための潜在的な説明は次のとおりです。未熟さ（生理学的成熟がないこと）、卵のクラッチを受精するために使用された後の精子のプラグの故障、と相手活性の欠如を。調査は女性のロブスターの交尾の成功を調べるため、この手法は、自然集団における生殖活動を文書化する分野で使用可能な信頼性の高いプロキシであることを示している。

<html>ここに報告されたデータは沿岸と沖合の場所の両方から発信され、女性ロブスターのサブセットでした。沿岸グループはCape Annの、マサチューセッツ州に近い沿岸海域で捕獲され、検査のための実験室にクーラーに輸送された44ロブスターで構成されていた。 186 offshoreロブスターの合計は、ジョージ銀行でキャプチャされ、ニューハンプシャー州における商業holding施設でサンプリングし?...

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

アメリカンロブスター漁業として利用甲殻類漁業への漁業による変化、潜在的に交尾の成功が低下する、生殖ダイナミクスに影響を与える可能性があります。この研究の目標は、生理学的または機能的に成熟したことが女性のロブスターの交尾の成功を確認するための非侵襲的方法を開発することでした。

非侵襲的方法<em&gt;その場で</em&gt;女性アメリカンロブスターにおける交配の成功（の決定<em&gt;ロブスターのamericanus</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 ビュー.  University of New Hampshire. アメリカンロブスター漁業として利用甲殻類漁業への漁業による変化、潜在的に交尾の成功が低下する、生殖ダイナミクスに影響を与える可能性があります。この研究の目標は、生理学的または機能的に成熟したことが女性のロブスターの交尾の成功を確認するための非侵襲的方法を開発することでした。 

ビデオ: 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.

ブナスケール昆虫への抵抗のためのアメリカのブナを（画面ための技術<em&gt;クリプトコッカスfagisuga</em&gt;リンド。）

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

環境学

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、のための改良INEX方法<em&gt;その場で</em&gt;微粒子と溶解した化合物の測定は、アクティブサスペンションフィーダによって処理します

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.

最適化された小型リン定量法を用いた粒状水酸化第二鉄に酸の吸着を確認する手順

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.

その場でパラフィン埋め込まれた大人サンゴの標本の交配の技術

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.

タバココナジラミの迅速同定のループ介在増幅法 (LAMP) アッセイ

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.

少年太平洋ヤツメウナギやアメリカのウナギの新しいマイクロ音響タグの注入

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

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.

純粋な落葉スタンドにおける3つの異なる方法を用いてリーフエリア指数推定

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 ...

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&#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.

植物環境相互作用のリアルタイム生理的フェノタイピングのためのテレメトリー、重力測定プラットフォーム

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.

ヤドリギ根絶剤 - ヤドリギの機械的および化学的同時制御のための新しいツール

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.

非侵襲的方法

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ブナスケール昆虫への抵抗のためのアメリカのブナを（画面ための技術

VacuSIP、のための改良INEX方法