この論文を構成して公開するための資金は、ミネソタ大学で昆虫学科にユスリカ研究グループ（LC Ferrington、ジュニア、PI）への複数の助成金や契約を通じて提供されました。 数字として使用されるフィールドワークの写真を共有するためのネイサン·ロバーツに感謝をこの原稿に関連したビデオで。

フィールドコレクション用品の調製<ol><li>研究デザインに基づいて収集し、各サンプルの1サンプル瓶（ 例えば、60ミリリットル）を取得する必要がありますSFPEサンプルの数を決定します。 </li><li>各サンプルジャーのための2つの日付と局所性のラベルを準備します。内側に1を配置し、瓶の外側に他を貼付。国、州、郡、市、水本体、GPS座標、日付、人の名前（複数可）のサンプルを収集：各日付および局所ラベルは、以下の情報が含まれていることを確認。 </li><li>他の具体的な資機材を（特定の材料/機器の表を参照）収集します。 </li></ol> 2.フィールドコレクション<ol><li>片手に幼虫トレイとその他でふるいを保持します。 SFPEが蓄積水に幼虫トレイを浸し（ 例えば、フォームの蓄積、思わぬ障害、緊急植生、バック破片、渦、銀行縁に沿って）（ 図2A）は 、ワットを許可しますえー、脱皮殻、および破片幼虫トレイを入力して、ふるいを通して、この材料を注ぎます。流水のシステムでのサンプリングは、サンプル範囲の下流端から始まり、（ 図2B）を上流作業する場合。静水システムでサンプリングした場合は、風下の海岸線から始まります。 <ol><li>各事前定義されたサンプルリーチ（一般的に100〜200のストリームから採取したサンプルのためのMが、水生監視サイトの全体の面積に依存する）内に10分間（又はそうでなければ特定のサンプリング政権に対して定義された）を繰り返し、ステップ2.1。適宜SFPE蓄積領域間を移動。 </li></ol></li><li>サンプルサイトからの水で満たされた噴出ボトルを使用して、ふるいの一つの領域にゴミを濃縮し、慎重に鉗子の援助と噴霧ボトルからのエタノールの流れで前標識されたサンプル瓶にSFPEサンプルを転送します。エタノールでサンプル瓶を埋めます。 </li><li>繰り返しは、すべてのサンプルについて2.1 2.2を繰り返します。 </li></ol> 3.サンプル·ピッキング注：このプロトコルの残りは300 SFPEのサブサンプルに関係し、他のサブサンプルのサイズに変更する必要があるかもしれません。勉強-具体的な目標とリソースを満たすためにSFPE方法を調整するためブシャールとFerringtonの9サブサンプリングとサンプリング周波数のガイドラインを参照してください。 <ol><li>各SFPEサンプルの1ドラムバイアルを割り当てます。各バイアルの内側に配置し、エタノールで完全バイアルを¾記入する日付と局所ラベルを準備します。 </li><li>対応するサンプル瓶から蓋を外し、添付の蛹脱皮殻をチェック。静かエタノールを充填した噴出ボトルを使用して、ペトリ皿に蓋オフ内容をすすぎます。探して、ピンセットを使用してサンプル瓶の内側からラベルを削除し、優しくペトリ皿の上にラベルから内​​容をすすぎます。脇のラベルを設定します。 </li><li>何SFPEは、サンプル瓶に残ってないことを確認するためにエタノールですすぎ、幼虫トレイにサンプル瓶の内容を転送します。蛹脱皮殻の一部を転送し、RESIペトリ皿のトレイから起因し、エタノール。サンプルをエタノールに覆われていることを確認してください。 </li><li>実体顕微鏡下でペトリ皿を置きます。体系蛹脱皮殻のためのペトリ皿の内容をスキャンします。ピンセットを用いて皿からすべての蛹脱皮殻を選択し、バイアル中に配置します。壊れている標本を選択しないで、乾燥させ、以降の識別の問題を回避するために圧縮（ すなわち、頭胸部と腹部の少なくとも半分を持っていません）。 注：いくつかのケースでは、属レベルの同定は、部分標本で可能かもしれませんが種を同定は、多くの場合、試料全体が存在することが必要です。 <ol><li>渦巻き皿と最初に検出されていない可能性があります皿の側面に貼り付けることができ、そのいずれかを含む追加の蛹脱皮殻、ならびに、任意の小さな半透明の試料をスキャンします。二つの連続スキャンが追加の蛹脱皮殻を明らかにしなくなるまで繰り返します。 </li></ol></li><li>繰り返しは、手順3.3と全部または300蛹脱皮殻まで3.4を拾ってきました。 300蛹脱皮殻を取り上げてきたとき、幼虫のトレイにペトリ皿から残渣を返し、エタノールでペトリ皿をすすぎます。そして、空のサンプル瓶に幼虫トレイから残留物を転送した日付と局所性のラベルを追加し、瓶の蓋を置きます。保持またはプロジェクト固有のプロトコルに応じて、残留​​物を処分します。 </li></ol> 4.サンプルソート<ol><li>すべてがちょうど標本をカバーするのに十分なエタノールで満たされたペトリ皿に標識されたバイアルから蛹脱皮殻を選んだ注ぎます。 </li><li>実体顕微鏡下で、独立した試験片の異なる形態学的基（ すなわち、morphotaxa）に、別々に標識されたバイアルに各morphotaxonを置くには、エタノールで完全番目の 3/4を満たしました。 <ol><li>ユスリカmorphotaxaを分離するために、外部形態的特徴を利用しています。例えば、頭胸部から、存在、大きさ、形状の違いを使用し、かつ頭結節、正面いぼ、正面毛、および胸部ホーンの着色。腹部から、morphotaxa分離（ 図4A）のための肛門ローブに加えて、棘、hookrows、シャグリーン、毛、および腹部のセグメントのスプリアスを使用します。追加の説明および形態学的特徴の数値のFerrington ら 5、Sæther26、ピンダーとReissの27を参照してください。 </li><li>試験片を乾燥し始める場合は、追加のエタノールを使用してください。 </li></ol></li></ol>取り付け5.スライド<ol><li> 95％エタノールで各morphotaxonためのマルチウェルプレートの1つのウェルを埋めます。 <ol><li>プレートの個々のウェルに取り付けられたスライドであることを各morphotaxonの複数の表現（ 例えば、全体の25％）を配置します。標本が十分に脱水するために、少なくとも10分間、十分に座ってできるようにします。 </li></ol></li><li>適切なサイト、コレクション、および識別informatiとラベルスライド（ 図3）に。 </li><li>実体顕微鏡のスライドを置きます。 注：ステージにテープで貼り付けたスライドのテンプレートは、一貫性のある配置のために有用です。 </li><li>スライド上のEuparalの低下を置き、それはカバースリップのサイズを近似するようEuparalを広げます。 Euparalで作業する場合、適切な換気を使用してください。 注：Euparalで作業する場合、適切な換気を使用してください。 </li><li>ピンセットを用いてEuparalに最初morphotaxonからの代表を埋め込みます。 注：試料から過剰のエタノールを無効にするには、ピンセットを用いて、穏やかに研究室に検体をタップ前Euparalでそれを埋め込むに拭き取ります。 </li><li>先の細いピンセットおよび/ ​​または解剖プローブ（ 図4A）を用いて、腹部から頭胸部を分離します。 <ol><li> ecdysial縫合糸（ 図4B）に沿って頭胸部を分割し、縫合糸の端が反対側（ 図4C）にあるように、頭胸部を開きます。 </li><li>オリエントCephalothorax腹側（ 図4C）が上を向いているように。 </li><li>腹部背側を配置し、すぐに頭胸部（ 図4C）の下に配置します。 </li></ol></li><li>試料上のカバーガラスを配置します。一方のエッジがスライドに触れると、角度でカバーガラスを保持し、その後徐々に低くし、気泡の形成を減少させるためにカバースリップをドロップします。試料を平らにカバースリップ上で軽く押し。 </li><li>繰り返して、すべての脱水の試料のために5.7を介して5.3を繰り返します。 </li></ol> 6.属の識別<ol><li>複合顕微鏡を用いてスライドに取り付けられた試験片の属を決定します。 ら 、Wiederholm 28とFerringtonでキーと診断を使用して属に検体を特定します。5。必要に応じて、 ら 、Ferringtonを使用して家族レベルの識別を確認する。5。注：Wiederholm 28とFerringtonので、多数の一般的な説明と改訂が行われています、ら 5は 、したがって、これらのキーと診断は不完全であり、主要な文献で ​​補充する必要があります。 </li></ol>

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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Communities in Freshwater Coastal Rock Pools of Lake Superior, with a Focus on Chironomidae (Diptera). , (2014).</li><li>Raunio, J., Heino, J., Paasivirta, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-Biting+Midges+in+Biodiversity+Conservation+and+Environmental+Assessment:+Findings+from+Boreal+Freshwater+Ecosystems.">Non-Biting Midges in Biodiversity Conservation and Environmental Assessment: Findings from Boreal Freshwater Ecosystems.</a> Ecological Indicators. 11, 1057-1064 (2011).</li><li>Kavanaugh, R. G., Egan, A. T., Ferrington, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+affecting+decomposition+rates+of+chironomid+(Diptera)+pupal+exuviae.">Factors affecting decomposition rates of chironomid (Diptera) pupal exuviae.</a> Chironomus: Newsletter on Chironomidae Research. 27, 16-24 (2014).</li><li>Anderson, A. M., Stur, E., Ekrem, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+and+Morphological+Methods+Reveal+Cryptic+Diversity+and+Three+New+Species+of+Nearctic+Micropsectra+(Diptera:+Chironomidae).">Molecular and Morphological Methods Reveal Cryptic Diversity and Three New Species of Nearctic Micropsectra (Diptera: Chironomidae).</a> Freshwater Science. 32, 892-921 (2013).</li><li>Ekrem, T., Willassen, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploring+Tanytarsini+Relationships+(Diptera:+Chironomidae)+using+Mitochondrial+COII+Gene+Sequences.">Exploring Tanytarsini Relationships (Diptera: Chironomidae) using Mitochondrial COII Gene Sequences.</a> Insect Systematics &amp; Evolution. 35, 263-276 (2004).</li><li>Ekrem, T., Willassen, E., Stur, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Comprehensive+DNA+Sequence+Library+is+Essential+for+Identification+with+DNA+Barcodes.">A Comprehensive DNA Sequence Library is Essential for Identification with DNA Barcodes.</a> Molecular Phylogenetics and Evolution. 43, 530-542 (2007).</li></ol>

著者は、彼らが競合する金融利害関係がないことを宣言します。

成功SFPEサンプル採取、スライドを取り付け、ピッキング並べ替え、および識別のための最も重要なステップは次のとおりです。（1）フィールドコレクション（ 図2A）の間、研究領域内の高SFPEの蓄積領域の位置を特定します。 （2）ゆっくりとサンプル中のすべてのピッキングSFPEの検出のためのペトリ皿の内容をスキャンします。 （3）（ 図4A）を取り付け、スライ?...

環境の健康を評価するための生物を使用し生物学的モニタリングプログラムは、多くの場合、水の品質を評価または生態系復元プログラムの成功を監視するために使用されます。底生大型無脊椎動物の集合体を使用して、簡易生物調査プロトコル（RBP）は1989年1ので、状態の水資源機関の間で人気となっている。このようなディップネット、Surberサンプラー、およびヘスサンプラ2としてRBPsために底生大型無脊椎動物をサンプリングする従来の方法、タイムすることができます高価な、消費、および、特定のマイクロハビタット3から集合体を測定することができます。特に水の体の生体情報を生成するための効率的な、代替RBPはユスリカ表面フローティング蛹脱皮殻（SFPE）3のコレクションを含みます。 ユスリカ（昆虫：双翅目）、一般的に非痛烈ユスリカとして知られているが、一般的に大人として浮上する前に、水生環境で発生する完全変態のハエです<html> 60;水の表面に。ユスリカファミリは、世界中に説明約5,000種と、種が豊富です。しかし、20,000人の種が4存在すると推定されます。ユスリカは、高い多様性と可変汚染許容レベル5の多くの水生生態系に水と生息環境の質を文書化するのに有用です。さらに、それらは一般的に、コミュニティ5,6で種の50％以上を占め、多くの場合、水系で最も豊富かつ広範底生大型無脊椎動物です。地上大人の出現に続いて、蛹脱皮殻（蛹皮膚をキャスト）は、水の表面に浮いたまま（ 図1）。蛹脱皮殻は、風や水流の作用を介して、銀行に沿って、または障害物の後ろに蓄積し、容易かつ迅速に前の24〜48時間の7時に登場したユスリカ種の総合的なサンプルを与えるために収集することができます。 ntent &quot;&gt;相対存在し、収集SFPEの分類学的組成物は他の人が5非常に敏感でありながら、いくつかの種は、非常に汚染寛容であることを考慮して、水質を反映SFPE法を含む従来の幼虫ユスリカサンプリング技術よりも多くの利点があります：（1）最小限の（2）サンプルは、生物を収集することに焦点を当てるのではなく、非生物肌なので、社会動態の軌跡には影響しませんしません;属〜（3）の識別、およびもしあれば、生息地の乱れがサンプリング領域で発生します多くの場合、種は、適切なキーおよび説明3与えられた比較的容易である;（4）の収集、処理、およびサンプルの識別は、従来のサンプリング方法3,8,9と比較して、効率的かつ経済的である;（5）累積脱皮殻から発信された分類群を表します微細環境10の広い範囲が、（6）の方法は、小川や河川、河口、LAK含め、ほぼすべての水生環境でも適用可能ですES、池、岩のプール、および湿地;彼らはすべての未成熟の段階を完了し、正常成人11として浮上している個人を表すので、（7）SFPEは多分生態系の健全性のより敏感な指標です。 SFPE方法はユスリカのコミュニティについての情報を収集するための新しいアプローチではありません。 SFPEの使用は、最初の1900年代初頭にティーネマン12によって示唆されました。様々な研究は、分類学上の調査（ 例えば、13〜15）、生物多様性と生態系の研究（ 例えば 7,16-19）、および生物学的評価（ 例えば、20〜22）のためにSFPEを使用しています。さらに、いくつかの研究は、サンプルデザイン、サンプルサイズ、及び種または属の様々な検出レベルを達成するために必要なサンプルのイベント（ 例えば、8,9,23）の数のさまざまな側面に対処しています。これらの研究は、種または属の比較的高い割合が緩やかefforで検出できることを示していますサンプル処理に関連したTまたは費用。例えば、アンダーソンとFerrington 8は 100カウントのサブサンプルに基づいて、1/3以下の時間がサンプルネットを浸漬するために比較SFPEサンプルを選択するために必要と判断しました。別の研究では、3-4 SFPEサンプルがソートされ、すべてのディップネットサンプルについてと種の豊富は、3の増加としてSFPEサンプルが検出種でディップネットのサンプルよりも効率的であったことが確認できたと判断しました。 SFPEサンプルは97.8％で、効率的な3であった例えば、15〜16種の種の豊かさの値を持つサイトで、平均ディップ正味効率は、45.7パーセントでした。 重要なことに、SFPE方法は生態学的評価のために、欧州連合24（ユスリカ蛹の脱皮殻法（CPET）として知られている）と北米25で標準化されているが、この方法は、詳細に記載されていません。 SFPE方法論の一つの用途は、 ら Ferrington、によって記述されました。 <s&gt; 3アップ。しかし、その研究の主な焦点は、SFPE方法の効率性、有効性、および経済性を評価することでした。この作業の目的は、サンプル収集、研究室処理、スライドマウントおよび属の識別を含む、詳細にSFPE方法のすべてのステップを記述することです。対象読者は、彼らの研​​究に伝統的な水質モニタリングの取り組みを拡大するに興味大学院生、研究者、専門家が含まれています。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Ethanol</td>
			<td>Fisher Scientific</td>
			<td>S25309B&#160;</td>
			<td>70-95%</td>
		</tr>
		<tr>
			<td>Plastic wash bottles</td>
			<td>Fisher Scientific</td>
			<td>0340923B</td>
			<td></td>
		</tr>
		<tr>
			<td>Sample jar</td>
			<td>Fisher Scientific</td>
			<td>0333510B</td>
			<td>Glass or plastic, 60-mL recommended</td>
		</tr>
		<tr>
			<td>Testing sieve</td>
			<td>Advantech</td>
			<td>120SS12F</td>
			<td>125-micron mesh size</td>
		</tr>
		<tr>
			<td>Larval tray</td>
			<td>BioQuip</td>
			<td>5524</td>
			<td>White</td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glass shell vials</td>
			<td>Fisher Scientific</td>
			<td>0333926B</td>
			<td>1-dram size</td>
		</tr>
		<tr>
			<td>Plastic dropper</td>
			<td>Thermo Scientific</td>
			<td>1371110</td>
			<td>30 to 35 drops/mL</td>
		</tr>
		<tr>
			<td>Fine forceps</td>
			<td>BioQuip</td>
			<td>4524</td>
			<td>#5</td>
		</tr>
		<tr>
			<td>Petri dish</td>
			<td>Carolina</td>
			<td>741158</td>
			<td>Glass or plastic</td>
		</tr>
		<tr>
			<td>Multi-well plate</td>
			<td>Thermo Scientific</td>
			<td align="right">144530</td>
			<td>Glass or plastic</td>
		</tr>
		<tr>
			<td>Glass microslides</td>
			<td>Thermo Scientific</td>
			<td>3010002</td>
			<td>3 x 1 in.</td>
		</tr>
		<tr>
			<td>Glass cover slips</td>
			<td>Thermo Scientific</td>
			<td>12-519-21G</td>
			<td>Circular or square</td>
		</tr>
		<tr>
			<td>Euparal mounting medium&#160;</td>
			<td>BioQuip</td>
			<td>6372B</td>
			<td></td>
		</tr>
		<tr>
			<td>Pigma pen</td>
			<td>BioQuip</td>
			<td>1154F</td>
			<td>Black</td>
		</tr>
		<tr>
			<td>Probe</td>
			<td>BioQuip</td>
			<td>4751</td>
			<td></td>
		</tr>
		<tr>
			<td>Kimwipes</td>
			<td>Kimberly-Clark Professional&#8482;</td>
			<td align="right">34120</td>
			<td></td>
		</tr>
	</tbody>
</table>

use of chironomidae (diptera) surface-floating pupal exuviae as a rapid bioassessment protocol for water bodies

底生大型無脊椎動物の集合体を使用して、簡易生物調査プロトコルが正常に水質に人間の影響を評価するために使用されています。残念ながら、そのようなディップネットなどの伝統的な底生幼虫のサンプリング方法は、時間がかかり、高価です。代替プロトコルは、ユスリカ表面フローティング蛹脱皮殻（SFPE）のコレクションを含みます。ユスリカは、未成熟の段階、通常、水生生息地で発生するハエ（双翅目）の種が豊富なファミリーです。大人ユスリカは、水から出てくる水の表面に浮い、彼らの蛹皮、または脱皮殻を残します。脱皮殻は、多くの場合、銀行に沿って、または、彼らはユスリカの多様性と豊かさを評価するために収集することができ、風や水流の作用によって、障害物の後ろに蓄積します。一部の種は他のものより汚染に対してより耐性があるので、ユスリカは、重要な生物学的指標として使用することができます。そのため、収集されたSFPEの相対量と種組成に反映します水質の変化。ここでは、フィールドコレクション、実験室処理、スライドマウントおよびユスリカSFPEの識別に関連する方法が詳細に記載されています。 SFPE方法の利点は、サンプリング領域、効率的かつ経済的なサンプル収集と研究室の処理、識別を容易に、ほぼすべての水生環境での適用性、および生態系のストレスの潜在的に、より高感度の尺度で最小の乱れがあります。この制限は、成人男性に関連付けられていない場合は種に蛹脱皮殻を識別するために、幼虫の生息の使用とできないことを決定することができないことがあります。

図1は、ユスリカのライフサイクルを示す図です。未成熟の段階（卵、幼虫、さなぎ）は、典型的に起こり、または密接に水生環境に関連します。幼虫ライフステージが完了すると、幼虫は、チューブ状のシェルターを構築し、周囲の基板にシルクの分泌物で自身を添付して蛹化が発生します。開発大人が成熟した後、蛹は、自分自身を解放し、大人が蛹脱皮殻から出てくること...

Watch this Scientific Journal Video about Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies at JoVE.com

Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

Rapid bioassessment protocols using benthic macroinvertebrates are often used to monitor and assess water quality. An efficient protocol involves collections of Chironomidae surface-floating pupal exuviae (SFPE). Here, techniques for field collection, laboratory processing, slide mounting, and identification of Chironomidae SFPE are described.

水域のためのユスリカ（双翅目）簡易生物調査プロトコルとして表面フローティング蛹脱皮殻の使用

Rapid bioassessment protocols using benthic macroinvertebrate assemblages have been successfully used to assess human impacts on water quality. ...

use-chironomidae-diptera-surface-floating-pupal-exuviae-as-rapid

Research

JoVE Journal

Environment

Use of Chironomidae (Diptera) Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

11.6K ビュー.  University of Minnesota. Rapid bioassessment protocols using benthic macroinvertebrates are often used to monitor and assess water quality. An efficient protocol involves collections of Chironomidae surface-floating pupal exuviae (SFPE). Here, techniques for field collection, laboratory processing, slide mounting, and identification of Chironomidae SFPE are described.

ビデオ: Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

Laboratory-determined Phosphorus Flux from Lake Sediments as a Measure of Internal Phosphorus Loading

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) loading from lake sediments can account for a substantial portion of the total P load in eutrophic, and some mesotrophic, lakes. Laboratory determination of P release rates from sediment cores is one approach for determining the role of internal P loading and guiding management decisions. Two principal alternatives to experimental determination of sediment P release exist for estimating internal load: in situ measurements of changes in hypolimnetic P over time and P mass balance. The experimental approach using laboratory-based sediment incubations to quantify internal P load is a direct method, making it a valuable tool for lake management and restoration.
Laboratory incubations of sediment cores can help determine the relative importance of internal vs. external P loads, as well as be used to answer a variety of lake management and research questions. We illustrate the use of sediment core incubations to assess the effectiveness of an aluminum sulfate (alum) treatment for reducing sediment P release. Other research questions that can be investigated using this approach include the effects of sediment resuspension and bioturbation on P release.
The approach also has limitations. Assumptions must be made with respect to: extrapolating results from sediment cores to the entire lake; deciding over what time periods to measure nutrient release; and addressing possible core tube artifacts. A comprehensive dissolved oxygen monitoring strategy to assess temporal and spatial redox status in the lake provides greater confidence in annual P loads estimated from sediment core incubations.

Lake eutrophication is a water quality issue worldwide, making the need to identify and control nutrient sources critical. Laboratory determination of phosphorus release rates from sediment cores is a valuable approach for determining the role of internal phosphorus loading and guiding management decisions.

内部リン負荷の尺度として湖堆積物の研究室に決定されたリンフラックス

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) ...

環境学

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.

腸内のメタンと反芻動物からの二酸化炭素排出量を監視するために、自動システム（GreenFeed）の使用

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

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most experiments examine only two or a few levels of CA concentration and a single soil type, but if CA can be varied as a gradient from subambient to superambient concentrations on multiple soils, we can discern whether past ecosystem responses may continue linearly in the future and whether responses may vary across the landscape. The Lysimeter Carbon Dioxide Gradient Facility applies a 250 to 500 &#181;l L-1 CA gradient to Blackland prairie plant communities established on lysimeters containing clay, silty clay, and sandy soils. The gradient is created as photosynthesis by vegetation enclosed in in temperature-controlled chambers progressively depletes carbon dioxide from air flowing directionally through the chambers. Maintaining proper air flow rate, adequate photosynthetic capacity, and temperature control are critical to overcome the main limitations of the system, which are declining photosynthetic rates and increased water stress during summer. The facility is an economical alternative to other techniques of CA enrichment, successfully discerns the shape of ecosystem responses to subambient to superambient CA enrichment, and can be adapted to test for interactions of carbon dioxide with other greenhouse gases such as methane or ozone.

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

The Lysimeter Carbon Dioxide Gradient Facility creates a 250 to 500 &#181;l L-1 linear carbon dioxide gradient in temperature-controlled chambers housing grassland plant communities on clay, silty clay, and sandy soil monoliths. The facility is used to determine how past and future carbon dioxide levels affect grassland carbon cycling.

CO<sub&gt; 2</subテスト用のCO&gt;濃度勾配施設<sub&gt; 2</sub&gt;濃縮および草地生態系の機能に及ぼす土壌の影響

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most ...

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

毒性のための飲料水試料の迅速な評価のためのRTgill-W1細胞とのインピーダンスベースの流体バイオチップの製造および試験

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

Protocol for Measuring the Thermal Properties of a Supercooled Synthetic Sand-water-gas-methane Hydrate Sample

Methane hydrates (MHs) are present in large amounts in the ocean floor and permafrost regions. Methane and hydrogen hydrates are being studied as future energy resources and energy storage media. To develop a method for gas production from natural MH-bearing sediments and hydrate-based technologies, it is imperative to understand the thermal properties of gas hydrates.
The thermal properties' measurements of samples comprising sand, water, methane, and MH are difficult because the melting heat of MH may affect the measurements. To solve this problem, we performed thermal properties' measurements at supercooled conditions during MH formation. The measurement protocol, calculation method of the saturation change, and tips for thermal constants' analysis of the sample using transient plane source techniques are described here.
The effect of the formation heat of MH on measurement is very small because the gas hydrate formation rate is very slow. This measurement method can be applied to the thermal properties of the gas hydrate-water-guest gas system, which contains hydrogen, CO2, and ozone hydrates, because the characteristic low formation rate of gas hydrate is not unique to MH. The key point of this method is the low rate of phase transition of the target material. Hence, this method may be applied to other materials having low phase-transition rates.

We present a protocol for measuring the thermal properties of synthetic hydrate-bearing sediment samples comprising sand, water, methane, and methane hydrate.

過冷却合成砂 - 水 - ガス・メタンハイドレート試料の熱特性を測定するためのプロトコル

Methane hydrates (MHs) are present in large amounts in the ocean floor and permafrost regions. Methane and hydrogen hydrates are being studied as future ...

Soil Lysimeter Excavation for Coupled Hydrological, Geochemical, and Microbiological Investigations

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled Earth-system processes. Such knowledge is imperative in improving predictions of hydro-biogeochemical cycles, especially under climate change scenarios. We present an experimental method, designed to capture sub-surface heterogeneity of an initially homogeneous soil system. This method is based on destructive sampling of a soil lysimeter designed to simulate a small-scale hillslope. A weighing lysimeter of one cubic meter capacity was divided into sections (voxels) and was excavated layer-by-layer, with sub samples being collected from each voxel. The excavation procedure was aimed at detecting the incipient heterogeneity of the system by focusing on the spatial assessment of hydrological, geochemical, and microbiological properties of the soil. Representative results of a few physicochemical variables tested show the development of heterogeneity. Additional work to test interactions between hydrological, geochemical, and microbiological signatures is planned to interpret the observed patterns. Our study also demonstrates the possibility of carrying out similar excavations in order to observe and quantify different aspects of soil-development under varying environmental conditions and scale.

This study presents an excavation method for investigating subsurface hydrological, geochemical, and microbiological heterogeneity of a soil lysimeter. The lysimeter simulates an artificial hillslope which was initially under homogeneous condition and had been subjected to approximately 5,000 mm of water over eight cycles of irrigation in an 18-month period.

結合水文学、地球化学、および微生物学的調査のための土壌ライシメータ発掘

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled ...

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

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

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

ろ過水のサンプルおよび環境DNAの抽出のためのフィルターカートリッジの使用

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

Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of scientific publications address microplastic pollution of the sea surface. The protocol below describes the methodology for sampling, sample preparation, separation and chemical identification of microplastic particles. A manta net fixed on an &#187;A frame&#171; attached to the side of the vessel was used for sampling. Microplastic particles caught in the cod end of the net were separated from samples by visual identification and use of stereomicroscopes. Particles were analyzed for their size using an image analysis program and for their chemical structure using ATR-FTIR and micro FTIR spectroscopy. The described protocol is in line with recommendations for microplastics monitoring published by the Marine Strategy Framework Directive (MSFD) Technical Subgroup on Marine Litter. This written protocol with video guide will support the work of researchers that deal with microplastics monitoring all over the world.

The protocol below describes the methodology for: microplastics sampling on the sea surface, separation of microplastic and chemical identification of particles. This protocol is in line with the recommendations for microplastics monitoring published by the MSFD Technical Subgroup on Marine Litter.

海面とサンプル分析にMicroplasticsサンプリングのためのプロトコル

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of ...

Measuring and Mapping Patterns of Soil Erosion and Deposition Related to Soil Carbonate Concentrations Under Agricultural Management

Spatial patterns of soil erosion and deposition can be inferred from differences in ground elevation mapped at appropriate time increments. Such changes in elevation are related to changes in near-surface soil carbonate (CaCO3) profiles. The objective is to describe a simple conceptual model and detailed protocol for repeatable field and laboratory measurements of these quantities. Here, accurate elevation is measured using a ground-based differential global positioning system (GPS); other data acquisition methods could be applied to the same basic method. Soil samples are collected from prescribed depth intervals and analyzed in the lab using an efficient and precise modified pressure-calcimeter method for quantitative analysis of inorganic carbon concentration. Standard statistical methods are applied to point data, and representative results show significant correlations between changes in soil surface layer CaCO3 and changes in elevation consistent with the conceptual model; CaCO3 generally decreased in depositional areas and increased in erosional areas. Maps are derived from point measurements of elevation and soil CaCO3 to aid analyses. A map of erosional and depositional patterns at the study site, a rain-fed winter wheat field cropped in alternating wheat-fallow strips, shows the interacting effects of water and wind erosion affected by management and topography. Alternative sampling methods and depth intervals are discussed and recommended for future work relating soil erosion and deposition to soil CaCO3.

Spatial patterns of soil erosion and deposition can be inferred from differences in ground elevation mapped at appropriate time increments. Such changes in elevation are related to changes in near-surface soil carbonates. Repeatable methods for field and laboratory measurements of these quantities and data analysis methods are described here.

測定と土壌侵食と堆積土炭酸濃度農業経営に関連のパターンのマッピング

Spatial patterns of soil erosion and deposition can be inferred from differences in ground elevation mapped at appropriate time increments. Such changes ...

A Simple Planting Technique for Re-establishing Trees Where Frequent Inundation Occurs

Many of the Everglades tree islands have lost elevation over the past century and most of their trees have died such that they are now covered with herbaceous plants. This protocol describes a simple, cost-effective tree planting technique needed for restoring degraded Everglade tree islands. The design is patterned after a natural Everglades process that creates floating peat islands, which allows tree survival and growth in flooded conditions and often leads to the development of tree islands. Commercially available peat bags were used as the medium for the growth and establishment of potted native tree saplings. The pop-up configuration floated initially and provided additional elevation to minimize inundation, with a single native tree species sapling and a single tree fertilizer spike. During a 3 year study involving 105 pop-ups, most plants survived (80%) and many thrived. Determining whether this technique can establish trees on a degraded tree island will require longer studies and extensive field tests.

This protocol describes a simple, cost-effective tree planting technique for restoring degraded Everglades tree islands that experience inundation. The design creates an island (pop-up) which floats initially and adds elevation to promote tree survival and growth under flooded conditions.

単純な頻繁に浸水が発生した再構築ツリー手法を植栽

Many of the Everglades tree islands have lost elevation over the past century and most of their trees have died such that they are now covered with ...