我々 はこれらの熱水合成実験の初期の方法論を開発するためアリゾナ州立大学の hog でグループを感謝し、特に、i. グールド、E. ショック、・ ウィリアムス、C. Glein、h. ハートネット、k. Fecteau、・ ロビンソンと C. Bockisch に感謝のガイダンスと参考に支援。Z のヤンと X. Fu Z のヤンにオークランド大学からスタートアップ資金によって資金を供給されました。

1. 熱水実験用サンプルを準備します。
<ol><li>水晶や石英ガラス管、例えば、 2 mm 内径 (ID) x 6 mm 外径 (OD) または ID x 12 が mm 外径 6 mm のサイズを選択し、有機物と使用する鉱物の量を定めます。この作品でニトロ ベンゼンと (例えば、 2 mm ID x 6 mm 外径) 石英管にロードする磁鉄鉱 (Fe3O4) の量は 3.0 μ L 13.9 mg それぞれです。 注: 大口径管材料の容易にロードするが管のシーリングのより多くの努力を必要とします。</li>
	<li>~ 30 cm と小さな断片にきれいな石英ガラス管をチューブ カッターを使用して長さにカットします。酸水素トーチを用いた適切な炎頭を閉じた管の一方の端をシールします。 注意: 酸水素のトーチを使用するため安全手順に従います。</li>
	<li>0.1 mg スケール (固体の場合) のバランスをとるし、秤量紙を用いた石英ガラス管に転送の開始の有機化合物の所定量の重量を量る。場合は、化合物は、液体 (例えば、この場合ニトロ ベンゼン)、小さな石英管にそれを転送する 1 マイクロリットル注射器 (例えば10 μ L) を使用します。パスツール ピペットで石英管に重量を量られた鉱物を追加し、脱・脱酸素水 (例えば、0.3 mL) を追加します。18.2 MΩ·cm 脱イオン水を使用し、超音波処理による deoxygenate。</li>
	<li>閉鎖弁付け真空ライン (~ 1 cm ID) に石英管を接続します。有機物と水が完全に冷凍されるまで 〜 3 分間液体窒素で満たされたデュワー フラスコで管を浸します。 注意: 転送し、液体窒素を使用するため安全手順に従います。</li>
	<li>チューブを液体窒素に浸漬に、ままとき真空バルブを開き、チューブのヘッドから空気を除去します。 注: このプロセスは、圧力が真空ポンプの圧力ゲージの 100 mtorr を下回るまで続くはず。</li>
	<li>バルブをオフ、液体窒素からチューブを削除し、室温まで温チューブ。ヘッド スペースに解決策から残りの空気の泡を解放する管の底を軽くたたきます。</li>
	<li>さらに 2 回の上記の凍結-融解性ポンプのサイクルを繰り返し、チューブのもう一方の端を密封する前に液体窒素でチューブを維持します。真空ラインを閉じ、閉じた全体の管を作る酸水素火炎を使用します。 注: チューブは、熱水実験を受け、液体の水膨張管のヘッド スペース台数が減少します。水の密度が 300 ° c 程度室温から約 30% を削減できるなど、計算し、管を密封した場合十分なヘッド ボリュームを残します。</li>
</ol>2. 熱水実験を設定します。
<ol><li>シールの手順後我慢シリカ チューブ小さなスチール パイプ (~ 30 cm の長さと直径 1.5 cm) に緩いねじキャップ、任意の圧力建物または管内管障害による損傷を防ぐために。</li>
	<li>よく恒温炉やオーブンでパイプを置き、所望の温度 (例えば、この仕事で 150 ° C) まで加熱します。水熱反応により温度を監視するのに炉内熱電対を使用します。</li>
	<li>(例えば、この作品で 2 h) 反応時間に達すると、すぐに、すばやく氷水風呂にパイプを置くことによって石英管を癒します。 注: 焼入プロセスは潜在的な逆行性反応を避け室温に冷却する 1 分未満かかります。</li>
</ol>3. 実験の後、サンプルを分析します。
<ol><li>チューブ カッターを使用して石英管を開き、パスツール ピペットを使用して 10 mL バイアルにすべての製品 (例えば、小さな石英管で ~0.3 mL) をすばやく転送します。</li>
	<li>ガスクロマトグラフィー (GC) 用内部標準として 8.8 mM ドデカンを含む 3 mL ジクロロ メタン (DCM) ソリューションとオーガニック製品を抽出します。キャップ バイアルとそれで手 2 分の渦 1 分間振る。 注: これはオーガニック有機相への抽出を容易にするのに役立ちます。また、製品の回復を確実に DCM の石英管の転送ピペットと内側の壁をすすいでください。ミネラル含量の高いサンプルより良い抽出の DCM ソリューションでそれらを超音波照射します。</li>
	<li>5 分使用して慎重に GC バイアルに DCM の層 (すなわち底層) から 〜 1 mL のサンプルを転送するパスツール ピペットの抽出液 (すなわちドデカンと DCM) に定住する鉱物粒子を許可します。</li>
	<li>ポリ毛細管カラム (例えば5 %diphenyl/95% ジメチルシロキサン) と炎イオン化検出器の GC を用いた有機製品の流通を分析します。50 ° c を開始、8 分間保持、220 ° C まで 10 ° C/分で増加し 10 分間保持、300 ° C まで 20 ° C/分で増加、300 ° C に 5 分セット インジェクター温度を保持するプログラムを GC オーブンを設定します。 注: を変更するために必要な GC プログラムは有機化合物の分析対象の種類に基づいています。</li>
	<li>GC 検を構築するには、試料の濃度と内部標準試料のピーク面積比をプロットします。</li>
	<li>前に、と後の反応、すなわち変換 % 有機原料の濃度に基づく反応換算 = ([初版]-[最終]) ⁄ [初期] × 100%。変換を使用すると、ミネラルを容易にまたは熱水有機変換が遅くかどうかを確認します。</li>
</ol>

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Catalysis by mineral surfaces.</a> Geochimica et Cosmochimica Acta. 58 (19), 4155-4177 (1994).</li><li>Yang, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrothermal+Photochemistry+as+a+Mechanistic+Tool+in+Organic+Geochemistry:+The+Chemistry+of+Dibenzyl+Ketone.">Hydrothermal Photochemistry as a Mechanistic Tool in Organic Geochemistry: The Chemistry of Dibenzyl Ketone.</a> The Journal of Organic Chemistry. 79 (17), 7861-7871 (2014).</li><li>Yang, Z., Hartnett, H. E., Shock, E. L., Gould, I. 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著者が明らかに何もありません。

本研究で我々 ニトロ ベンゼン ミネラル磁鉄鉱として使用例熱水有機反応における鉱物の影響を評価するのに方法をデモンストレーションします。小さな石英ガラス管で、実験を行い、再現性の高い結果がマグネタイトの実験、すなわち30.3 ± 1.4% ニトロ ベンゼンの変換、有効性と信頼性のあるこれを示唆しているので観察、熱水のプロトコル。なし鉱物の実験でニトロ ベンゼンの変換は 5.2 ± 2.1%、ミネラルの実験より低再現性を示します。サンプルで使用される小さな管の μ L (または mg) を考慮した原料の低変換なし鉱物実験で不確実性の比較的高い可能性があります。低変換反応の再現性を高めるためには、大きい内部ボリュームを持つシリカ チューブが示唆されました。サンプルの量は限られている、または化学物質のコストが高い場合、このプロトコルは小規模な実験のために特に有用かもしれない。このプロトコルでは、鉱物と非鉱物の熱水合成実験を実施できます。
前述したように、この熱水プロトコルに反応管、安易な操作方法、低または僅かの触媒効果11,14の低コストなど、他の伝統的な方法で特定の利点はします。しかし、限られた鉱物の強度と安定性のため石英管が故障原因 450 ° C 以上の温度または圧力 400 バー15、上で長時間熱水実験の臨界点以上の近くに適してしない可能性があります。水。このメソッドのもう一つの制限かもしれないということです、高温 (例えば、 > 400 ° C) で水晶も解散は、有機の熱水反応を妨害する溶解性シリカの種を作り出すことができます。シリカの溶解は、液の pH によって、影響を受ける可能性があります。、ので塩、酸や塩基の存在、生存温度は純粋な水システムのそれよりも低くなる可能性が、これらの要因は、高温においても考慮する必要があります。実験。さらに、ゴールドなど柔軟な原子炉材料と比較して、シリカ チューブは、通常いくつかの気相反応が発生することができる外部の圧力を適用することによって縮小できないヘッド ボリュームに関連付けられます。
さらに、シリカ チューブ内の液体の体積は、実験の成功を決定する際に重要な可能性があります。SUPCRT9216を使用して熱力学計算に基づいて、たとえば、(P土) 水の飽和蒸気圧力は 300 ° c 以上 85 バーを達することができる、シリカ チューブ内の液体水の量は 30% によって拡張できます。高温高圧で生き残るために厚い石英ガラス管 (内径/外径比すなわち、 < 0.3) 大きいヘッド スペースを使用する必要があります。同じ直径でもさまざまなメーカーからシリカ チューブ異なる温度での失敗があります。したがって、温度及び圧力抑制シリカ チューブは使用する前に徹底的にテストする必要がありますの種類ごとに。この熱水のプロトコルからそのホウケイ酸ガラスを除外すると、反応は、通常、300 ° C の上の温度を扱うことができないので注意してください。また、ある有機化合物を読み込み「スティッキー」または狭いシリカ チューブに粘性かもしれない挑戦、(例えば、 6 mm ID x 12 mm 外径) 大口径チューブが推奨される場合。

熱水環境 (すなわち、高温高圧の水溶液) 地球上至るところでされます。有機化合物の水熱化学は、有機堆積盆地、石油タンク、地下生物圏1,2,3などの地球化学的設定の広い範囲で重要な役割を果たしています。熱水系における有機炭素変換だけでなく地球豊富なミネラルなど溶存または固体の無機材料、でも純粋な水溶液で発生します。劇的に、選択的に影響を与える各種の有機化合物の水熱反応性鉱物が発見されているが、1,4、5複雑な熱水系におけるミネラルの効果を識別する方法課題として残っています。本研究の目的は、熱水有機反応に及ぼす鉱物を研究するための比較的簡単な実験プロトコルを提供することです。
水熱反応の実験的研究は伝統的にゴールド、チタンまたはステンレス鋼6,7,8,9から成っている堅牢な原子炉を使用します。たとえば、金の袋またはカプセル使用されている好意的、金は柔軟性ができ、サンプルの圧力、外部から水を加圧によって制御するサンプル中の気相の生成を避けますため。ただし、これらの原子炉は高価であり、潜在的な金属の触媒効果10と関連付けることができます。したがって、これらの熱水実験用低コスト、高い信頼性と別の方法を見つけることが不可欠です。
近年、水晶や石英ガラス製反応管は熱水実験11,12,13に多く適用されています。貴重な金やチタンと比較して、水晶や石英ガラスはまた強い材料がかなり安いです。もっと重大に、石英管ほとんどの触媒効果を示しているし、不活性にすることができます水熱反応11,14金として。このプロトコルではシリカの厚肉管の小規模の熱水有機鉱物実験を行うための一般的方法をについて説明します。デモンストレーションとしてミネラル効果を示すために 150 ° C の熱水溶液中の鉄酸化物鉱物 (すなわち磁鉄鉱) の有無でモデル化合物 (すなわち、ニトロ ベンゼン) を用いた例実験を提案する、このメソッドの有効性。

<table>
	<tbody>
		<tr>
			<td>Chemicals:</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dichloromethane</td>
			<td>VWR</td>
			<td>BDH23373.400</td>
			<td></td>
		</tr>
		<tr>
			<td>Dodecane</td>
			<td>Sigma-Aldrich</td>
			<td>297879</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrobenzene</td>
			<td>Sigma-Aldrich</td>
			<td>252379</td>
			<td></td>
		</tr>
		<tr>
			<td>Fe2O3</td>
			<td>Sigma-Aldrich</td>
			<td>310050</td>
			<td></td>
		</tr>
		<tr>
			<td>Fe3O4</td>
			<td>Sigma-Aldrich</td>
			<td>637106</td>
			<td></td>
		</tr>
		<tr>
			<td>Supplies:</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Silica tube</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum pump</td>
			<td>WELCH</td>
			<td>2546B-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum line</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Oven</td>
			<td>Hewlett Packard</td>
			<td>5890</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermocouple</td>
			<td>BENETECH</td>
			<td>GM1312</td>
			<td></td>
		</tr>
		<tr>
			<td>Gas chromatography</td>
			<td>Agilent</td>
			<td>7820A</td>
			<td></td>
		</tr>
	</tbody>
</table>

an experimental protocol for studying mineral effects on organic hydrothermal transformations

土地、および深海の熱水噴出孔に間欠泉、温泉などの熱水環境下での有機・無機相互作用が広く行われます。ミネラルの役割は、多くの熱水の有機地球化学的プロセスで重要です。ゴールド、チタン、プラチナ、またはステンレス スチール製の原子炉を使用して含まれています、伝統的な熱水方法論は高コストまたは好ましくない金属触媒効果に通常関連付けられます。最近では、熱水実験で費用対効果や不活性の水晶や石英ガラス管を使用して成長傾向があります。ここでは、我々 は石英管内有機鉱物の熱水合成実験を遂行するためプロトコルを提供し、試料、実験装置、製品の分離と定量分析に不可欠な手順について述べる。モデル有機化合物、ニトロ ベンゼンを使用して特定の熱水条件下での分解に鉄を含む鉱物、マグネタイトの効果を表示する実験を紹介します。この手法は、比較的簡単な実験室システムにおける複雑な有機鉱物熱水相互作用に適用できます。

このアプローチを使用して熱水の有機・無機相互作用モデル化合物を用いた簡単な実験を研究する方法を示すためには、ニトロ ベンゼン、ミネラル磁鉄鉱 (Fe3O4) 150 ° C と 5 の水熱条件で行った2 h のバー。ミネラルの効果を見る、ミネラルなしのニトロ ベンゼンの実験も同じ熱水条件下で実施しました。図 1 aのように、2 つのシリカ チューブは熱水実験前にプロトコル、次を作られました。透明な鉱物と封管とマグネタイト管展示中の黒色鉱物。ニトロ ベンゼンの開始濃度が両方の 0.1 M (0.3 mL の脱イオン水を脱酸素化) と追加の磁鉄鉱 13.9 mg であった。熱水処理後鉱物管変化が認められなかった色、磁鉄鉱とチューブと化した茶色の色 (図 1 b)、磁鉄鉱からヘマタイト (Fe2O3) への酸化反応を意味するのに対し。ガスクロマト グラフ分析に基づき、磁鉄鉱の影響 (図 2) 実験間ニトロ ベンゼンの変換によって明らかにされました。いいえ鉱物実験でニトロ ベンゼンの計算された変換は 5.2%。ただし、磁鉄鉱の存在下でニトロ ベンゼン変換 6 の要因によって増加した 30.3% であった。さらに、重複するが、独立した実験を行い、それぞれ 2.1%、1.4% いいえ鉱物およびマグネタイトの実験のために 1 つの標準偏差を求めた (図 2)。これらの結果は酸化還元反応によっておそらく、マグネタイトを提案する、水熱条件を与えられたでニトロ ベンゼンの反応を大幅促進することができます。このプロトコルは、鉱物の影響下で熱水有機劣化の定量化における再現性の比較的高い成功する発見されました。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58230/58230fig1.jpg"> 図 1: 磁鉄鉱の存在の有無でニトロ例実験。熱水実験; の前に (、) 石英ガラス管(b) 熱水実験後の石英ガラス管。磁鉄鉱石英管で色の変化があることに注意してください。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58230/58230fig1large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58230/58230fig2.jpg"> 図 2: 150 の ° C および 5 バーで熱水条件下での 2 時間後ニトロ ベンゼン変換の実験反応変換、ニトロ ベンゼンの量によって計算されます、反応後の反応します。誤差には、重複した実験の平均の 1 つの標準偏差があります。いいえ鉱物およびマグネタイト実験の違いは明確にニトロ ベンゼンの熱分解に関するミネラルの効果を示します。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58230/58230fig2large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。

Watch this Scientific Journal Video about An Experimental Protocol for Studying Mineral Effects on Organic Hydrothermal Transformations at JoVE.com

An Experimental Protocol for Studying Mineral Effects on Organic Hydrothermal Transformations

地球-豊富なミネラルは、天然の熱水系における重要な役割を果たします。ここでは、我々 は熱水条件下での有機・無機相互作用の実験的調査のため信頼性が高く、コスト効果の高い方法をについて説明します。

有機水熱変換のミネラル効果を研究するための実験プロトコル

Organic-mineral interactions are widely occurring in hydrothermal environments, such as hot springs, geysers on land, and the hydrothermal vents in the ...

an-experimental-protocol-for-studying-mineral-effects-on-organic

Nanyang Technological University

Research

JoVE Journal

Environment

5.6K ビュー.  Oakland University. 地球-豊富なミネラルは、天然の熱水系における重要な役割を果たします。ここでは、我々 は熱水条件下での有機・無機相互作用の実験的調査のため信頼性が高く、コスト効果の高い方法をについて説明します。

ビデオ: An Experimental Protocol for Studying Mineral Effects on Organic Hydrothermal Transformations

Experimental Protocol for Manipulating Plant-induced Soil Heterogeneity

Coexistence theory has often treated environmental heterogeneity as being independent of the community composition; however biotic feedbacks such as plant-soil feedbacks (PSF) have large effects on plant performance, and create environmental heterogeneity that depends on the community composition. Understanding the importance of PSF for plant community assembly necessitates understanding of the role of heterogeneity in PSF, in addition to mean PSF effects. Here, we describe a protocol for manipulating plant-induced soil heterogeneity. Two example experiments are presented: (1) a field experiment with a 6-patch grid of soils to measure plant population responses and (2) a greenhouse experiment with 2-patch soils to measure individual plant responses. Soils can be collected from the zone of root influence (soils from the rhizosphere and directly adjacent to the rhizosphere) of plants in the field from conspecific and heterospecific plant species. Replicate collections are used to avoid pseudoreplicating soil samples. These soils are then placed into separate patches for heterogeneous treatments or mixed for a homogenized treatment. Care should be taken to ensure that heterogeneous and homogenized treatments experience the same degree of soil disturbance. Plants can then be placed in these soil treatments to determine the effect of plant-induced soil heterogeneity on plant performance. We demonstrate that plant-induced heterogeneity results in different outcomes than predicted by traditional coexistence models, perhaps because of the dynamic nature of these feedbacks. Theory that incorporates environmental heterogeneity influenced by the assembling community and additional empirical work is needed to determine when heterogeneity intrinsic to the assembling community will result in different assembly outcomes compared with heterogeneity extrinsic to the community composition.

Understanding the role of environmental heterogeneity in species coexistence has typically focused on types of heterogeneity that are extrinsic to the community&#8217;s species composition. We provide novel detailed methods for creating soil heterogeneity treatments using soils subject to plant-soil feedback conditioning, or heterogeneity intrinsic to the community composition.

の操作植物に誘導される土壌の不均一性のための実験プロトコール

Coexistence theory has often treated environmental heterogeneity as being independent of the community composition; however biotic feedbacks such as ...

環境学

Evaluation of Integrated Anaerobic Digestion and Hydrothermal Carbonization for Bioenergy Production

Lignocellulosic biomass is one of the most abundant yet underutilized renewable energy resources. Both anaerobic digestion (AD) and hydrothermal carbonization (HTC) are promising technologies for bioenergy production from biomass in terms of biogas and HTC biochar, respectively. In this study, the combination of AD and HTC is proposed to increase overall bioenergy production. Wheat straw was anaerobically digested in a novel upflow anaerobic solid state reactor (UASS) in both mesophilic (37 &#176;C) and thermophilic (55 &#176;C) conditions. Wet digested from thermophilic AD was hydrothermally carbonized at 230 &#176;C for 6 hr for HTC biochar production. At thermophilic temperature, the UASS system yields an average of 165 LCH4/kgVS&#160;(VS: volatile solids) and 121 L CH4/kgVS&#160;at mesophilic AD over the continuous operation of 200 days. Meanwhile, 43.4 g of HTC biochar with 29.6 MJ/kgdry_biochar was obtained from HTC of 1 kg digestate (dry basis) from mesophilic AD. The combination of AD and HTC, in this particular set of experiment yield 13.2 MJ of energy per 1 kg of dry wheat straw, which is at least 20% higher than HTC alone and 60.2% higher than AD only.

A novel Upflow Anaerobic Solid State (UASS) reactor was used for biogas production from fibrous feedstock. Digestate from UASS reactor was hydrothermally carbonized into HTC biochar in a pressurized batch reactor. The integration of the two bioenergy concepts was applied in this study to increase overall bioenergy production.

バイオエネルギー生産のための統合された嫌気性消化と水熱炭化の評価

Lignocellulosic biomass is one of the most abundant yet underutilized renewable energy resources. Both anaerobic digestion (AD) and hydrothermal ...

Exploring the Effects of Atmospheric Forcings on Evaporation: Experimental Integration of the Atmospheric Boundary Layer and Shallow Subsurface

Evaporation is directly influenced by the interactions between the atmosphere, land surface and soil subsurface. This work aims to experimentally study evaporation under various surface boundary conditions to improve our current understanding and characterization of this multiphase phenomenon as well as to validate numerical heat and mass transfer theories that couple Navier-Stokes flow in the atmosphere and Darcian flow in the porous media. Experimental data were collected using a unique soil tank apparatus interfaced with a small climate controlled wind tunnel. The experimental apparatus was instrumented with a suite of state of the art sensor technologies for the continuous and autonomous collection of soil moisture, soil thermal properties, soil and air temperature, relative humidity, and wind speed. This experimental apparatus can be used to generate data under well controlled boundary conditions, allowing for better control and gathering of accurate data at scales of interest not feasible in the field. Induced airflow at several distinct wind speeds over the soil surface resulted in unique behavior of heat and mass transfer during the different evaporative stages.

A protocol for the design and construction of a soil tank interfaced to a small climate controlled wind tunnel to study the effects of atmospheric forcings on evaporation is presented. Both the soil tank and wind tunnel are instrumented with sensor technologies for the continuous in situ measurement of environmental conditions.

蒸発させる上で大気強制力の影響を探る：大気境界層と浅い地下の実験的インテグレーション

Evaporation is directly influenced by the interactions between the atmosphere, land surface and soil subsurface. This work aims to experimentally study ...

Experimental Protocol for Detecting Cyanobacteria in Liquid and Solid Samples with an Antibody Microarray Chip

Global warming and eutrophication make some aquatic ecosystems behave as true bioreactors that trigger rapid and massive cyanobacterial growth; this has relevant health and economic consequences. Many cyanobacterial strains are toxin producers, and only a few cells are necessary to induce irreparable damage to the environment. Therefore, water-body authorities and administrations require rapid and efficient early-warning systems providing reliable data to support their preventive or curative decisions. This manuscript reports an experimental protocol for the in-field detection of toxin-producing cyanobacterial strains by using an antibody microarray chip with 17 antibodies (Abs) with taxonomic resolution (CYANOCHIP). Here, a multiplex fluorescent sandwich microarray immunoassay (FSMI) for the simultaneous monitoring of 17 cyanobacterial strains frequently found blooming in freshwater ecosystems, some of them toxin producers, is described. A microarray with multiple identical replicates (up to 24) of the CYANOCHIP was printed onto a single microscope slide to simultaneously test a similar number of samples. Liquid samples can be tested either by direct incubation with the antibodies (Abs) or after cell concentration by filtration through a 1- to 3-&#956;m filter. Solid samples, such as sediments and ground rocks, are first homogenized and dispersed by a hand-held ultrasonicator in an incubation buffer. They are then filtered (5 - 20 &#956;m) to remove the coarse material, and the filtrate is incubated with Abs. Immunoreactions are revealed by a final incubation with a mixture of the 17 fluorescence-labeled Abs and are read by a portable fluorescence detector. The whole process takes around 3 h, most of it corresponding to two 1-h periods of incubation. The output is an image, where bright spots correspond to the positive detection of cyanobacterial markers.

Experimental Protocol for Detecting Cyanobacteria in Liquid and Solid Samples with an Antibody Microarray Chip

The presence of cyanobacterial toxins in fresh water reservoirs for human consumption is a major concern for water management authorities. To evaluate the risk of water contamination, this article describes an protocol for the in-field detection of cyanobacterial strains in liquid and solid samples by using an antibody microarray chip.

検出するための実験プロトコール<em&gt;シアノバクテリア</em&gt;抗体マイクロアレイチップと液体と固体試料中の

Global warming and eutrophication make some aquatic ecosystems behave as true bioreactors that trigger rapid and massive cyanobacterial growth; this has ...

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 ...

Protocol for Assessing the Relative Effects of Environment and Genetics on Antler and Body Growth for a Long-lived Cervid

Cervid phenotype can be placed into one of two categories: efficiency, which promotes survival over extravagant morphometric growth, and luxury, which promotes growth of large weaponry and body size. Populations of the same species display each phenotype depending on environmental conditions. Although antler and body size of male white-tailed deer (Odocoileus virginianus) varies by physiographic region in Mississippi, USA and is strongly correlated with regional variation in nutritional quality, the effects of population-level genetics from native stocks and previous re-stocking efforts cannot be disregarded. This protocol describes how we designed a controlled study, where other factors that influence phenotype, such as age and nutrition, are controlled. We brought wild-caught pregnant females and six-month-old fawns from three distinct physiographic regions in Mississippi, USA to the Mississippi State University Rusty Dawkins Memorial Deer Unit. Deer from the same region were bred to produce a second generation of offspring, allowing us to assess generational responses and maternal effects. All deer ate the same high-quality (20% crude protein deer pellet) diet ad libitum. We uniquely marked each neonate and recorded body mass, hind foot, and total body length. Each subsequent fall, we sedated individuals via remote injection and sampled the same morphometrics plus antlers of adults. We found that all morphometrics increased in size from first to second generation, with full compensation of antler size (regional variation no longer present) and partial compensation of body mass (some evidence of regional variation) evident in the second generation. Second generation males that originated from our poorest quality soil region displayed about a 40% increase in antler size and about a 25% increase in body mass when compared to their wild harvested counterparts. Our results suggest phenotypic variation of wild male white-tailed deer in Mississippi are more related to differences in nutritional quality than population-level genetics.

Phenotypic differences among cervid populations may be related to population-level genetics or nutrition; discerning which is difficult in the wild. This protocol describes how we designed a controlled study where nutritional variation was eliminated. We found that phenotypic variation of male white-tailed deer was more limited by nutrition than genetics.

アントラーとシカの長命の体の成長の遺伝と環境の相対的な効果の評価のためのプロトコル

Cervid phenotype can be placed into one of two categories: efficiency, which promotes survival over extravagant morphometric growth, and luxury, which ...

Experimental Column Setup for Studying Anaerobic Biogeochemical Interactions Between Iron (Oxy)Hydroxides, Trace Elements, and Bacteria

Fate and speciation of trace elements (TEs), such as arsenic (As) and mercury (Hg), in aquifers are closely related to physio-chemical conditions, such as redox potential (Eh) and pH, but also to microbial activities that can play a direct or indirect role on speciation and/or mobility. Indeed, some bacteria can directly oxidize As(III) to As(V) or reduce As(V) to As(III). Likewise, bacteria are strongly involved in Hg cycling, either through its methylation, forming the neurotoxin monomethyl mercury, or through its reduction to elemental Hg&#176;. The fates of both As and Hg are also strongly linked to soil or aquifer composition; indeed, As and Hg can bind to organic compounds or (oxy)hydroxides, which will influence their mobility. In turn, bacterial activities such as iron (oxy)hydroxide reduction or organic matter mineralization can indirectly influence As and Hg sequestration. The presence of sulfate/sulfide can also strongly impact these particular elements through the formation of complexes such as thio-arsenates with As or metacinnabar with Hg.
Consequently, many important questions have been raised on the fate and speciation of As and Hg in the environment and how to limit their toxicity. However, due to their reactivity towards aquifer components, it is difficult to clearly dissociate the biogeochemical processes that occur and their different impacts on the fate of these TE.
To do so, we developed an original, experimental, column setup that mimics an aquifer with As- or Hg-iron-oxide rich areas versus iron depleted areas, enabling a better understanding of TE biogeochemistry in anoxic conditions. The following protocol gives step by step instructions for the column set-up either for As or Hg, as well as an example with As under iron and sulfate reducing conditions.

Experimental Column Setup for Studying Anaerobic Biogeochemical Interactions Between Iron OxyHydroxides, Trace Elements, and Bacteria

Fate and speciation of arsenic and mercury in aquifers are closely related to physio-chemical conditions and microbial activity. Here, we present an original experimental column setup that mimics an aquifer and enables a better understanding of trace element biogeochemistry in anoxic conditions. Two examples are presented, combining geochemical and microbiological approaches.

(オキシ) 鉄の水酸化物、微量元素、細菌の嫌気性生物地球化学的相互作用を研究するため実験的列の設定

Fate and speciation of trace elements (TEs), such as arsenic (As) and mercury (Hg), in aquifers are closely related to physio-chemical conditions, such as ...

Collection and Extraction of Occupational Air Samples for Analysis of Fungal DNA

Traditional methods of identifying fungal exposures in occupational environments, such as culture and microscopy-based approaches, have several limitations that have resulted in the exclusion of many species. Advances in the field over the last two decades have led occupational health researchers to turn to molecular-based approaches for identifying fungal hazards. These methods have resulted in the detection of many species within indoor and occupational environments that have not been detected using traditional methods. This protocol details an approach for determining fungal diversity within air samples through genomic DNA extraction, amplification, sequencing, and taxonomic identification of fungal internal transcribed spacer (ITS) regions. ITS sequencing results in the detection of many fungal species that are either not detected or difficult to identify to species level using culture or microscopy. While these methods do not provide quantitative measures of fungal burden, they offer a new approach to hazard identification and can be used to determine overall species richness and diversity within an occupational environment.

Determining the fungal diversity within an environment is a method utilized in occupational health studies to identify health hazards. This protocol describes DNA extraction from occupational air samples for amplification and sequencing of fungal ITS regions. This approach detects many fungal species that can be overlooked by traditional assessment methods.

コレクションと真菌 DNA の分析のための産業大気サンプルの抽出

Traditional methods of identifying fungal exposures in occupational environments, such as culture and microscopy-based approaches, have several ...

Experimental Protocol for Detecting Mitochondrial Function in Hepatocytes Exposed to Organochlorine Pesticides

This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by environmental organochlorine pesticides (OCPs) in hepatocytes. HepG2 cells were exposed to &#946;-hexachlorocyclohexane (&#946;-HCH) for 24 h at an equivalent dose of internal exposure in general population. Ultrastructure in hepatocytes was examined by transmission electron microscopy (TEM) to show the damage of mitochondria. Mitochondrial function was further evaluated by mitochondrial fluorescence intensity, adenosine 5'-triphosphate (ATP) levels, oxygen consumption rate (OCR) and mitochondrial membrane potential (MMP) in HepG2 cells incubated with &#946;-HCH. The mitochondria fluorescence intensity after stained by mitochondrial green fluorescent probe was observed with a fluorescence microscopy. The luciferin-luciferase reaction was used to determine ATP levels. The MMP was detected by the cationic dye JC-1 and analyzed under flow cytometry. OCR was measured with an extracellular flux analyzer. In summary, these protocols were used in detecting mitochondrial function in hepatocytes with to investigate mitochondria damages.

Understanding the influence of environmental organochlorine pesticides (OCPs) on mitochondrial function in hepatocytes is important in exploring the mechanism of OCPs causing metabolic disorders. This paper presents detailed methods on detecting hepatic mitochondrial function.

有機塩素系農薬に曝露した肝細胞におけるミトコンドリア機能の検出実験プロトコル

This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by ...

The Hawaii Protocol for Scientific Monitoring of Coffee Berry Borer: a Model for Coffee Agroecosystems Worldwide

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at capturing and quantifying the dynamics and impact of this invasive insect pest as well as the development of its host plant across a heterogeneous landscape. The cornerstone of this comprehensive monitoring system is timely georeferenced data collection on CBB movement, coffee berry infestation, mortality by the fungus Beauveria bassiana, and coffee plant phenology via a mobile electronic data recording application. This electronic data collection system allows field records to be georeferenced through built-in global positioning systems, and is backed by a network of weather stations and records of farm management practices. Comprehensive monitoring of CBB and host plant dynamics is an essential part of an area-wide project in Hawaii to aggregate landscape-level data for research to improve management practices. Coffee agroecosystems in other parts of the world that experience highly variable environmental and socioeconomic factors will also benefit from implementing this protocol, in that it will drive the development of customized integrated pest management (IPM) to manage CBB populations.

Comprehensive monitoring of coffee berry borer and host plant dynamics is essential for aggregating landscape-level data to improve management of this invasive pest. Here, we present a protocol for scientific monitoring of coffee berry borer movement, infestation, mortality, coffee plant phenology, weather, and farm management via a mobile electronic data recording application.

コーヒーのベリーの穴あけ器の科学的なモニタリングのためのハワイ プロトコル: コーヒー生態系世界のモデル

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at ...