著者に感謝 Pi 芬、林、氏魏肺・ ツェン、ミス プリンスエド ワード島玄林博士甲陸チュアン フィールド サンプリングとラボ分析と「CATNET」の開発・実用化のための支援乗組員とボード上の調査船海洋研究技術者の支援-I およびサンプリング遠征中に海洋研究 II は大歓迎します。この作品は台湾省科学によって部分的サポートされて、技術付与 104-2611-M-002-019, 97-3114-M-002-006 96-2611-M-002-004 95-2611-M-002-009 91-2611-M-002-007。この原稿は、温輝リー嬢の彼女の巨大な献身と台湾の海洋の研究への貢献のためのメモリに書き込まれます。

次のプロトコルには、有害な化学物質の使用が含まれます。ください安全性データ シート (SDS) を熟読し、制度化学物質の安全性ガイドラインに従ってください。
1. 多層重力ろ過サンプラー準備
<ol><li>
クリーニングのサンプラー
	<ol>
<li>アニオン性プロテアーゼ酵素洗剤の 1% (w/v) と、チューブとろ過の単位を記入し、浸し 24 h. フラッシュ逆浸透膜二重蒸留水 (RO DDW) 多層重力ろ過サンプラー徹底的に 0.1% (v/v) でそれを埋める塩酸 (HCl、試薬グレード)、72 時間浸漬。</li>
		<li>徹底的に逆浸透ダブル蒸留純水 (RO-DD-DIW) 3 〜 5 倍 20 リットル以上の時間とビニール袋に群集を格納と多層重力ろ過サンプラーをフラッシュします。</li>
	</ol>
</li>
	<li>
粒子サンプル コンテナー洗浄/準備
	<ol>
<li>粒子のコンテナーとして低密度ポリエチレン (LDPE、125 mL) またはフッ素化エチレン プロピレン (FEP、125 mL) ボトルを使用します。それぞれ最初アルカリ性洗剤 (マイクロ、1%) の 50% (v/v) 硝酸 (HNO3、試薬グレード)、それから少なくとも 24、48、24 h の 10% (v/v) HCl ソリューションでそれらを浸すことによってボトルをきれい。脱イオン水 (RO-DD-DIW) 2 つの浸漬手順間でボトルをすすいでください。</li>
		<li>最終的な HCl 浸漬後、徹底的に脱イオン水 (RO DD DIW) でボトルを洗浄、クリーン ルーム、クラス 100 のクリーン ベンチの中にボトルを乾燥します。 多層重力ろ過サンプラ洗浄瓶を添付またはきれいにシールは輸送のためそれらを PE ジッパー袋、二重袋のボトルします。</li>
	</ol>
</li>
	<li>
多層重力ろ過サンプラーの群集
	<ol>
<li>サンプラーの上に 6 方向入口に (0.635 cm 外径) 6 4 m 長い化学的に抵抗力がある熱可塑性エラストマー チューブを接続します。</li>
		<li>低密度ポリエチレン サンプル容器 (LDPE 125 mL) を 3 つの異なるメッシュ ナイロン フィルターをクリーン ルーム外に配置されている 10 μ m のメッシュ フィルターをクリーンアップです (下記参照) 後、(ベンチ) の順序で組み立てる 63 μ m メッシュ フィルターで、中高 153 μ m メッシュの内側にフィルターを実行します。輸送、ポリエチレン (PE) 袋の 2 つの層の多層重力ろ過サンプラーを保存し、ポリプロピレン (PP) 出荷用コンテナーに配置。</li>
	</ol>
</li>
</ol>2. サンプリング
<ol><li>
サンプル コレクション
	<ol>
<li>サンプリング サイトに到着すると、調査船の甲板上のコンテナーから多層重力ろ過サンプラーを外し、サンプラーと袋を開ける人があります。その後、PE 手袋を入れて、高いマルチ ボトル サンプリング アレイ上 6 20 L PTFE 被覆サンプリング ボトルの水栓に 6 の 4 m 熱可塑性エラストマー チューブを接続し、このろ過装置に海水を導くことがあります。海水を通過方向の入り江と、粒子/プランクトンが優しく分離/分別ネットを通じて、網の基地で保護される LDPE ボトル 125 mL に落ち着きます。</li>
		<li>海水には (沿岸海水用通常 120 L と外洋水 240 L) が流れて後の順序で各ネットを削除 (まず、153 μ m、63 μ m と 10 μ m 最後に) クラス 100 のクリーン ベンチで、微量金属きれい 0.4 μ m f ネットをスプレーiltered 海水洗浄すべてのプランクトン ネットの内側表面に付着します。125 mL ペットボトルで高濃度粒子/プランクトンを海水を収集します。</li>
		<li>ネットからこれらのボトルを緩めてフィルター酸洗浄吸引ろ過装置を通して再び集中粒子/プランクトンとソリューションの事前の重量を量った、酸洗浄 47 mm の低真空下で 10 μ m 孔サイズのポリカーボネート フィルター条件 (< 5 kPa)。</li>
		<li>その後、サンプラーを流れる 5 L PE 容器内の水の 2 〜 5 L を収集し、酸洗浄吸引ろ過装置でこれらのサンプル水をフィルター海水の少なくとも 20 L を待つ 10 μ m より小さい粒子/プランクトンを収集するにはあらかじめ重量を量られた、酸洗浄、47 mm、0.4 μ m 孔サイズのポリカーボネート フィルター。</li>
		<li>真空ろ過後高純度 DDW 水海水、海の塩の粒子/プランクトンの乾燥重量の決定に及ぼす影響を最小限に抑えることの残基を削除するサンプル フィルターを直ちに洗浄します。壊れやすいプランクトンを損傷を防ぐためにミリリットルのカップルだけにすすぎのボリュームを維持します。</li>
		<li>、この洗浄のステップ後慎重に削除しフィルター真空ろ過装置から、酸洗浄、あらかじめ重量を量られたアクリル プラスチック シャーレ、にサンプル フィルターを格納、ジッパーのついたビニール袋に密封します。さらにサンプル前処理および化学分析のための土地ベースの研究室に戻るまで-20 ° C のフリーザー搭載で袋をしてください。</li>
	</ol>
</li>
</ol>3. サンプル治療
<ol><li>
凍結乾燥と粒子の消化
	<ol>
<li>凍結乾燥機のコレクターの部屋で粒子のサンプル フィルターを置き、マシンをオンにします。機械温度-40 ° C に達すると、機械の真空ポンプをオンに、凍結乾燥プロセスを開始します。 注: 真空度は、0.12 mBar 以下着実に維持する必要があります。くださいユーザー マニュアルを注意深く読み、各ステップの製造元のガイドラインに従ってください。</li>
		<li>72 時間後をオフに凍結乾燥機、乾燥フィルターを削除し、それらの重量を量る。あらかじめ重量を量られたパーフルオロアルコキシ アルカン (PFA) 容器 (容量 60 mL) に乾燥のサンプル フィルターを配置し、船2,3,6,7に集中している超高純度硝酸 3 mL を加えます。</li>
		<li>2.5 kg ・ m、定トルクをトルクレンチで血管を強化し、最初の消化シーケンスの 12 h 130 ° C で従来のオーブンで血管を配置します。冷却した後、血管をオーブンから取り出し、船、船2,3,6,7に超高純度フッ化水素酸 2 mL を追加します。</li>
		<li>2.5 kg ・ m のトルクで血管を強化し、2 番目の消化シーケンスである 12 h 130 ° C で従来のオーブンで血管を配置します。冷却した後、船、船2,3,6,7に 4.5% 純度ホウ酸液 16 mL を追加します。</li>
		<li>2.5 キロ メートルの一定トルクの血管を締め、最終消化順序の 12 h 130 ° C のオーブンでサンプルを消化します。冷却した後、各船の重さし、最終的な質量と各消化ソリューション最終 digestant ボリュームを生成するための具体的な重量を決定します。 注: 特定の質量は、digestant のまさに 1.00 mL の重量を測定によって決定されます。</li>
		<li>慎重に、digestant をさらに微量金属分析用 30 mL 酸洗浄 PE ボトルに注ぐ。</li>
	</ol>
</li>
	<li>
微量金属分析
	<ol>
<li>微量金属元素濃度 (Cd、Cu、Fe、Mn、Ni、および Al) 消化ソリューションの黒鉛炉原子吸光分析計 (吸)6を使用して粒子を決定します。</li>
		<li>精度のテストとして、カナダ国立研究評議会の国立標準研究所の技術から河口域底質標準物質から海底堆積物試料などの標準物質 (CRM) を使用して、米国および欧州委員会の科学と知識のサービスからプランクトン参考資料。プロセスは、CRM、微量金属の認定値の 95% に 107% 回復を与えます。</li>
	</ol>
</li>
</ol>

<ol>
	<li>Jeandel, C., 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=What+did+we+learn+about+ocean+particle+dynamics+in+the+GEOSECS-JGOFS+era.">What did we learn about ocean particle dynamics in the GEOSECS-JGOFS era.</a> Progr. Oceanogr. 133, 6-16 (2015).</li><li>Lam, P., 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=Methods+for+analyzing+the+concentration+and+speciation+of+major+and+trace+elements+in+marine+particles.">Methods for analyzing the concentration and speciation of major and trace elements in marine particles.</a> Progr. Oceanogr. 133, 32-42 (2015).</li><li>Collier, R., Edmond, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+trace+element+geochemistry+of+marine+biogenic+particulate+matter.">The trace element geochemistry of marine biogenic particulate matter.</a> Progr. Oceanogr. 13, 113-199 (1984).</li><li>Donat, J. R., Bruland, K. W., Steinnes, E., Salbu, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+elements+in+the+oceans.">Trace elements in the oceans.</a> Trace Elements in Natural Waters. , 247-280 (1995).</li><li>Wen, L. -. S., Santschi, P., Tang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction+between+radioactively+labeled+colloids+and+natural+particles:+evidence+for+colloidal+pumping.">Interaction between radioactively labeled colloids and natural particles: evidence for colloidal pumping.</a> Geochim. Cosmochim. Ac. 61, 2867-2878 (1997).</li><li>Wen, L. -. S., Warnken, K., Santschi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+organic+carbon,+iron,+and+aluminium+oxyhydroxides+as+trace+metal+carriers:+Comparison+between+the+Trinity+River+and+the+Trinity+River+Estuary+(Galveston+Bay,+Texas).">The role of organic carbon, iron, and aluminium oxyhydroxides as trace metal carriers: Comparison between the Trinity River and the Trinity River Estuary (Galveston Bay, Texas).</a> Mar. Chem. 112, 20-37 (2008).</li><li>Hurd, D., Spencer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marine+particles:+analysis+and+characterization.">Marine particles: analysis and characterization.</a> American Geophysical Union. , (1991).</li><li>Martin, J. H., Knauer, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+elemental+composition+of+plankton.">The elemental composition of plankton.</a> Geochim. Cosmochim. Ac. 37, 1639-1653 (1973).</li><li>Morel, F., Price, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biogeochemical+cycles+of+trace+metals+in+the+oceans.">The biogeochemical cycles of trace metals in the oceans.</a> Science. 300, 944-947 (2003).</li><li>Ho, T. -. 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Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+metal+composition+of+size-fractionated+plankton+in+the+Western+Philippine+Sea:+the+impact+of+anthropogenic+aerosol+deposition.">Trace metal composition of size-fractionated plankton in the Western Philippine Sea: the impact of anthropogenic aerosol deposition.</a> Limnol Oceanogr. , (2017).</li><li>Grasshoff, K., Kremling, K., Ehrhardt, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods of seawater analysis. , (2007).</li></ol>

共同著者氏アラン荘は特許の共同所有者と興味を持つユーザーのコレクション装置を製造 (中国楽器株式) 会社のゼネラル マネージャー。特許は、9th、2015年125 月に終わった。

中に検体の採取、処理、前処理、分析の目的で細心の注意が必要です取得の信頼性の高いトレース プランクトンと一般的に非常に低濃度で存在している、天然水中の懸濁粒子の金属濃度汚染を低減します。そのため、設計およびサンプリング装置、サンプル容器、材料を準備する手順を収集する、プロセスのサンプルが海洋環境中の微量金属の高品質データ取得に向けたすべての重要な手順...

海洋中の粒子は、海洋物質循環1で重要な役割を果たします。サイズ、鉱物組成などの粒子の特性の最も深く 1 つの地質や前浜の設定から別の2変更できます。加えて、海洋における元素の分布が海洋植物プランクトンのライフ サイクルに関連付けられている: 成長、死、沈没、赤潮3,4。海洋の粒子にまたがるサブミクロン粒子から大きな凝集体に至るまでのサイズの少なくとも 4 桁 (> 5 mm)。ほとんどの粒子は、ウイルスの換散、滲出、分泌、糞便のペレット製造などのプロセスから派生生物学的。その他の粒子は、細胞、細胞の残骸や隆起材料1の物理的な凝固から形成されます。両方の物質循環と生物学的現象と粒子4,5,6内で粒子の様々 な化学的および生物学的特性を制御します。これらの粒子は動物プランクトンや saprotrophs など、いくつかの有機体のための食糧源と同様、重要な生息地です。したがって、粒子の運命は頻繁に生物学的過程と粒子の周りによって変更することができます彼らのサイズに関連します。
海洋の粒子を通常サンプリング、濾過がこのアプローチは海洋の粒子の構成内容と同種ではないので、粒子の特性を識別するに特定のあいまいさを紹介します。主体がほぼ完全に懸濁液、小型・低密度粒子の懸濁粒子が懸濁液のより大きくより密度の高い粒子の変化量と流体条件によっては、時間の短い期間にのみ混在します。7. プランクトン試料の微量金属の組成の最初のレポートは研究容器8プランクトン トウまたは中断するプランクトン ネットで収集されました。著者はしばしば金属粒子を発見し、ペイント チップ サンプルの化学分析のための海洋の粒子のサンプリング時の汚染の深刻な問題を示唆しています。その他、純ゴムいかだによって曳航または塩化ビニル (PVC) を使用して-手のウィンチ3。粒子の信頼性の高いサンプリングの難しさ、特に微量元素より困難海洋粒子の化学組成の私達の理解の進歩になります。そのため、植物プランクトン中の微量元素濃度に関する最も重要な情報は、文化研究9,10から来ています。このような認識、海洋科学者が過去 30 年間11上海で粒子を研究するための新しいメソッドを作成する動機となった。
海洋学者は、その場でろ過、船上濾過など様々 なサンプリング技術を使用しているし、セジメント トラップ11。特に外洋と深海粒子濃度が非常に低いであるため、非汚染のサンプルを収集する海水の大量の処理に挑戦することができます (0.001 〜 0.1 mg/L)。大量の海水微量金属元素濃度を測定するパーティクルの十分な量を取得するフィルターを適用する必要です。一部の研究者は、沈降粒子から粒子を分離するのにサイズ分画測定法を使用しています。しかし、粒径、空隙率、密度、および図形は、すべて影響粒子沈降速度をことができます。セジメント トラップは、それらが沈降粒子設計されているため、懸濁粒子を収集するために実用的なツールではありません。したがって、最小の汚染懸濁粒子の十分な量を集めることができるサンプリング ・治療法を開発することが重要です。したがって、その場で濾過により分画のサイズはまだ海洋のサンプリング ツールボックスで有望なツールそれは海洋の粒子のダイナミクスに関する重要な情報を明らかにすることができますので。ここでは、正常にテスト述べる微量金属をきれい、ポリテトラフルオロ エチレン (PTFE) から 1 つのパスでボード上の海水のサンプリング装置、大容量 (120-240 L) を扱うことができる多層重力ろ過コーティングで水のサンプリング ボトル、マルチ ボトル サンプリング配列。この捕集装置が順番に酸洗浄した合成ナイロン ネットを使用し、ネットが優しくサイズ分画された懸濁物質と植物プランクトン12,13,を収集するポリカーボネート容器に含まれている14,15 (図 1)。この作業の目的は、金属粒子の関連付けと海洋環境における反応ダイナミクスを研究するためより良いツールを提供し、さまざまなプランクトン、粒子、およびこれらの微量金属の運命の私達の理解を改善環境。

<table>
	<tbody>
		<tr>
			<td>thermoplastic elastomer (C-Flex) Tubings</td>
			<td>Cole Palmer</td>
			<td>EW-06424-67</td>
			<td>O.D. 0.635 cm, Opaque White 1/8&#34;ID x 1/4&#34;OD, 25 ft/pack</td>
		</tr>
		<tr>
			<td>LDPE Bottle (Nalgene)</td>
			<td>ThermoFisher Scientific</td>
			<td>2103-0004</td>
			<td>125 mL, Nalgene Wide-Mouth LDPE Bottles with Closure</td>
		</tr>
		<tr>
			<td>anionic protease enzyme detergent detergent (Tergazyme)</td>
			<td>Alconox</td>
			<td>1104-1</td>
			<td>1&#215;4 lb box (1.8 kg)</td>
		</tr>
		<tr>
			<td>Hydrochloric Acid</td>
			<td>Sigma-Aldrich</td>
			<td>258148</td>
			<td>Reagent grade</td>
		</tr>
		<tr>
			<td>Nitric acid</td>
			<td>Sigma-Aldrich</td>
			<td>695025</td>
			<td>Reagent grade</td>
		</tr>
		<tr>
			<td>alkaline detergnet (Micro)</td>
			<td>Cole Palmer</td>
			<td>EW-99999-14</td>
			<td>Micro-90 Cleaning Solution</td>
		</tr>
		<tr>
			<td>polycarbonate filter, 47 mm, 0.4 &#181;m</td>
			<td>Sigma-Aldrich</td>
			<td>WHA111107</td>
			<td>Whatman Nuclepore Track-Etched Membranes, diam. 47 mm, pore size 0.4 &#956;m, polycarbonate</td>
		</tr>
		<tr>
			<td>polycarbonate filter, 47 mm, 10 &#181;m</td>
			<td>Sigma-Aldrich</td>
			<td>WHA111115</td>
			<td>Whatman Nuclepore Track-Etched Membranes, diam. 47 mm, pore size 10 &#956;m, polycarbonate</td>
		</tr>
		<tr>
			<td>PFA vessel, 60 ml capacity</td>
			<td>Savillex</td>
			<td>300-060-03</td>
			<td>60 mL Digestion Vessel, Flat Interior, Flat Exterior, Buttress Threaded Top</td>
		</tr>
		<tr>
			<td>Nitric acid, ultrapure</td>
			<td>Seastar Chemicals</td>
			<td>N/A</td>
			<td>BASELINE Nitric Acid</td>
		</tr>
		<tr>
			<td>HF, ultrapure</td>
			<td>Seastar Chemicals</td>
			<td>N/A</td>
			<td>BASELINE Hydrofluoric Acid</td>
		</tr>
		<tr>
			<td>Boric acid, ultrapure</td>
			<td>Seastar Chemicals</td>
			<td>N/A</td>
			<td>BASELINE Hydrobromic Acid</td>
		</tr>
		<tr>
			<td>polyethylene (PE) gloves</td>
			<td>Safty Zone</td>
			<td>GDPL-MD-5</td>
			<td>Clear Powder Free Polyethylene Gloves</td>
		</tr>
		<tr>
			<td>Multiple layer filtering and collecting device</td>
			<td>Sino Instrumnets Co. Ltd</td>
			<td>not available</td>
			<td>Multiple layer filtering and collecting device, CATNET</td>
		</tr>
		<tr>
			<td>10 um Nylon filters, Nitex</td>
			<td>Dynamic Aqua-Supply Ltd.</td>
			<td>NTX 10</td>
			<td>Nitex - Standard Widths (40 - 44 inches)</td>
		</tr>
		<tr>
			<td>60 um Nylon filters, Nitex</td>
			<td>Dynamic Aqua-Supply Ltd.</td>
			<td>NTX 60</td>
			<td>Nitex - Standard Widths (40 - 44 inches)</td>
		</tr>
		<tr>
			<td>150 um Nylon filters, Nitex</td>
			<td>Dynamic Aqua-Supply Ltd.</td>
			<td>NTX 150</td>
			<td>Nitex - Standard Widths (40 - 44 inches)</td>
		</tr>
		<tr>
			<td>torque wrench</td>
			<td>Halfords</td>
			<td>200238</td>
			<td>Halfords Professional Torque Wrench 8-60Nm</td>
		</tr>
		<tr>
			<td>multi-bottle sampling array, Rosette</td>
			<td>General Oceanics</td>
			<td>Model 1018</td>
			<td>Rosette Sampler</td>
		</tr>
		<tr>
			<td>PTFE-coated sampling bottles, GO-Flo</td>
			<td>General Oceanics</td>
			<td>108020T</td>
			<td>GO-Flo water sampler teflon coated</td>
		</tr>
		<tr>
			<td>Marine sediment reference materials</td>
			<td>National Research Council Canada</td>
			<td>MESS-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Estuarine sediment standard reference material</td>
			<td>National Institute of Standards and Technology</td>
			<td>1646a</td>
			<td></td>
		</tr>
		<tr>
			<td>Plankton reference material</td>
			<td>The European Commission&#39;s science and knowledge service</td>
			<td>CRM414</td>
			<td></td>
		</tr>
	</tbody>
</table>

an ultra-clean multilayer apparatus for collecting size fractionated marine plankton and suspended particles

海で多くの微量元素の分布、成長、死、および海洋プランクトンの赤潮と中断された沈降粒子に強く関連付けられています。すべてプラスチック (ポリプロピレン、ポリカーボネート) 多層ろ過システム海で浮遊粒子状物質 (SPM) のコレクションを紹介します。この超クリーン サンプリング デバイスを設計されており、微量元素の研究用に開発されました。すべての非金属材料の細心の選択、プロシージャ フローをインラインでの使用率は、サンプリング中に可能な金属汚染を最小限に抑えます。このシステムは、正常にテストされており、沿岸や外洋海域でさまざまな大きさの粒子における微量元素 (例えばFe、Al、Mn、Cd、Cu、Ni) を決定するために調整します。サウス イースト アジア時系列 (席) 駅南シナ海からの結果は、日内変動と有光層におけるプランクトンの分布することができます簡単に解決し、認識ことを示します。台湾海峡の海面でサイズ分画された粒子の化学分析は示唆大きい粒子 (> 153 μ m) 小さい粒子 (10 63 μ m) が主無機物質から成る間、得られた大抵生物学的。Cd から離れて増加するサイズの金属 (Fe、Al、Mn, Cu, Ni) の濃度が減少しました。

近代海洋学の発展に伴い、それは「クリーン技術」を使用する一般的な練習では今海洋粒子やプランクトンの金属濃度の正確なトレースを取得します。天然水の粒子の大半は μ g/L の範囲に低 mg/l なので海水の大量の治療は周囲の環境で様々 な粒子の微量金属元素の地球化学的および生物学的効果を検討する必要。きれいな、多層重力ろ過 (「CATNET」) のサンプリング...

Watch this Scientific Journal Video about An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles at JoVE.com

An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles

サイズ分画した海洋プランクトンや懸濁粒子を収集するための超きれいな積層装置

プランクトンや懸濁粒子は、海の物質循環に大きな役割を果たします。ここでは、海水の大量の処理能力を持つ、超クリーン、低応力法の粒子と海でプランクトンの様々 なサイズのコレクションを提供します。

The distributions of many trace elements in the ocean are strongly associated with the growth, death, and re-mineralization of marine plankton and those ...

an-ultra-clean-multilayer-apparatus-for-collecting-size-fractionated

Research

JoVE Journal

Environment

8.7K ビュー.  National Taiwan University. プランクトンや懸濁粒子は、海の物質循環に大きな役割を果たします。ここでは、海水の大量の処理能力を持つ、超クリーン、低応力法の粒子と海でプランクトンの様々 なサイズのコレクションを提供します。

ビデオ: An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles

A Fish-feeding Laboratory Bioassay to Assess the Antipredatory Activity of Secondary Metabolites from the Tissues of Marine Organisms

Marine chemical ecology is a young discipline, having emerged from the collaboration of natural products chemists and marine ecologists in the 1980s with the goal of examining the ecological functions of secondary metabolites from the tissues of marine organisms. The result has been a progression of protocols that have increasingly refined the ecological relevance of the experimental approach. Here we present the most up-to-date version of a fish-feeding laboratory bioassay that enables investigators to assess the antipredatory activity of secondary metabolites from the tissues of marine organisms. Organic metabolites of all polarities are exhaustively extracted from the tissue of the target organism and reconstituted at natural concentrations in a nutritionally appropriate food matrix. Experimental food pellets are presented to a generalist predator in laboratory feeding assays to assess the antipredatory activity of the extract. The procedure described herein uses the bluehead, Thalassoma bifasciatum, to test the palatability of Caribbean marine invertebrates; however, the design may be readily adapted to other systems. Results obtained using this laboratory assay are an important prelude to field experiments that rely on the feeding responses of a full complement of potential predators. Additionally, this bioassay can be used to direct the isolation of feeding-deterrent metabolites through bioassay-guided fractionation. This feeding bioassay has advanced our understanding of the factors that control the distribution and abundance of marine invertebrates on Caribbean coral reefs and may inform investigations in diverse fields of inquiry, including pharmacology, biotechnology, and evolutionary ecology.

This bioassay employs a model predatory fish to assess the presence of feeding-deterrent metabolites from organic extracts of the tissues of marine organisms at natural concentrations using a nutritionally comparable food matrix.

海洋生物の組織からの二次代謝産物のAntipredatory活性を評価するために魚の供給研究所バイオアッセイ

Marine chemical ecology is a young discipline, having emerged from the collaboration of natural products chemists and marine ecologists in the 1980s with ...

環境学

A Protocol for Collecting and Constructing Soil Core Lysimeters

Leaching of nutrients from land applied fertilizers and manure used in agriculture can lead to accelerated eutrophication of surface water. Because the landscape has complex and varied soil morphology, an accompanying disparity in flow paths for leachate through the soil macropore and matrix structure is present. The rate of flow through these paths is further affected by antecedent soil moisture. Lysimeters are used to quantify flow rate, volume of water and concentration of nutrients leaching downward through soils. While many lysimeter designs exist, accurately determining the volume of water and mass balance of nutrients is best accomplished with bounded lysimeters that leave the natural soil structure intact. 
Here we present a detailed method for the extraction and construction of soil core lysimeters equipped with soil moisture sensors at 5 cm and 25 cm depths. Lysimeters from four different Coastal Plain soils (Bojac, Evesboro, Quindocqua and Sassafras) were collected on the Delmarva Peninsula and moved to an indoor climate controlled facility. Soils were irrigated once weekly with the equivalent of 2 cm of rainfall to draw down soil nitrate-N concentrations. At the end of the draw down period, poultry litter was applied (162 kg TN ha-1) and leaching was resumed for an additional five weeks. Total recovery of applied irrigation water varied from 71% to 85%. Nitrate-N concentration varied over the course of the study from an average of 27.1 mg L-1 before litter application to 40.3 mg L-1 following litter application. While greatest flux of nutrients was measured in soils dominated by coarse sand (Sassafras) the greatest immediate flux occurred from the finest textured soil with pronounced macropore development (Quindocqua).

A detailed method for extraction and assembly of intact soil core lysimeters and their use for study of leachate and associated loss of nutrients from surface applied poultry litter is demonstrated.

土壌コアLysimetersを収集し、構築するためのプロトコル

Leaching of nutrients from land applied fertilizers and manure used in agriculture can lead to accelerated eutrophication of surface water. Because the ...

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 ...

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

水生生物に浮遊堆積物の影響を評価するためのフロースルー露光装置

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

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 ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment during manufacturing and transport. Because of their environmental persistence, they are widely distributed in the oceans and on beaches all over the world. They can act as a vector of potentially toxic organic compounds (e.g., polychlorinated biphenyls) and might consequently negatively affect marine organisms. Their possible impacts along the food chain are not yet well understood. In order to assess the hazards associated with the occurrence of plastic pellets in the marine environment, it is necessary to develop methodologies that allow for rapid determination of associated organic contaminant levels. The present protocol describes the different steps required for sampling resin pellets, analyzing adsorbed organochlorine pesticides (OCPs) and identifying the plastic type. The focus is on the extraction of OCPs from plastic pellets by means of a pressurized fluid extractor (PFE) and on the polymer chemical analysis applying Fourier Transform-InfraRed (FT-IR) spectroscopy. The developed methodology focuses on 11 OCPs and related compounds, including dichlorodiphenyltrichloroethane (DDT) and its two main metabolites, lindane and two production isomers, as well as the two biologically active isomers of technical endosulfan. This protocol constitutes a simple and rapid alternative to existing methodology for evaluating the concentration of organic contaminants adsorbed on plastic pieces.

Microplastics act as vector of potentially toxic organic contaminants with unpredictable effects. This protocol describes an alternative methodology for assessing the levels of organochlorine pesticides adsorbed on plastic pellets and identifying the polymer chemical structure. The focus is on pressurized fluid extraction and attenuated total reflectance Fourier transform infrared spectroscopy.

プラスチックペレットからの有機塩素系農薬の抽出と塑性解析

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment ...

Preparation of Authigenic Pyrite from Methane-bearing Sediments for In Situ Sulfur Isotope Analysis Using SIMS

Different sulfur isotope compositions of authigenic pyrite typically result from the sulfate-driven anaerobic oxidation of methane (SO4-AOM) and organiclastic sulfate reduction (OSR) in marine sediments. However, unravelling the complex pyritization sequence is a challenge because of the coexistence of different sequentially formed pyrite phases. This manuscript describes a sample preparation procedure that enables the use of secondary ion mass spectroscopy (SIMS) to obtain in situ &#948;34S values of various pyrite generations. This allows researchers to constrain how SO4-AOM affects pyritization in methane-bearing sediments. SIMS analysis revealed an extreme range in &#948;34S values, spanning from -41.6 to +114.8&#8240;, which is much wider than the range of &#948;34S values obtained by the traditional bulk sulfur isotope analysis of the same samples. Pyrite in the shallow sediment mainly consists of 34S-depleted framboids, suggesting early diagenetic formation by OSR. Deeper in the sediment, more pyrite occurs as overgrowths and euhedral crystals, which display much higher SIMS &#948;34S values than the framboids. Such 34S-enriched pyrite is related to enhanced SO4-AOM at the sulfate-methane transition zone, postdating OSR. High-resolution in situ SIMS sulfur isotope analyses allow for the reconstruction of the pyritization processes, which cannot be resolved by bulk sulfur isotope analysis.

Preparation of Authigenic Pyrite from Methane-bearing Sediments for In Situ Sulfur Isotope Analysis Using SIMS

Analyses of the sulfur isotopic composition (&#948;34S) of pyrite from methane-bearing sediments have typically focused on bulk samples. Here, we applied secondary ion mass spectroscopy to analyze the &#948;34S values of various pyrite generations to understand the diagenetic history of pyritization.

シムズを用いた硫黄同位体分析における原位置メタン胚胎堆積物から自生黄鉄鉱の準備

Different sulfur isotope compositions of authigenic pyrite typically result from the sulfate-driven anaerobic oxidation of methane (SO4-AOM) and ...

Development of New Methods for Quantifying Fish Density Using Underwater Stereo-video Tools

The use of video camera systems in ecological studies of fish continues to gain traction as a viable, non-extractive method of measuring fish lengths and estimating fish abundance. We developed and implemented a rotating stereo-video camera tool that covers a full 360 degrees of sampling, which maximizes sampling effort compared to stationary camera tools. A variety of studies have detailed the ability of static, stereo-camera systems to obtain highly accurate and precise measurements of fish; the focus here was on the development of methodological approaches to quantify fish density using rotating camera systems. The first approach was to develop a modification of the metric MaxN, which typically is a conservative count of the minimum number of fish observed on a given camera survey. We redefine MaxN to be the maximum number of fish observed in any given rotation of the camera system. When precautions are taken to avoid double counting, this method for MaxN may more accurately reflect true abundance than that obtained from a fixed camera. Secondly, because stereo-video allows fish to be mapped in three-dimensional space, precise estimates of the distance-from-camera can be obtained for each fish. By using the 95% percentile of the observed distance from camera to establish species-specific areas surveyed, we account for differences in detectability among species while avoiding diluting density estimates by using the maximum distance a species was observed. Accounting for this range of detectability is critical to accurately estimate fish abundances. This methodology will facilitate the integration of rotating stereo-video tools in both applied science and management contexts.

We describe a new method for counting fishes, and estimating relative abundance (MaxN) and fish density using rotating stereo-video camera systems. We also demonstrate how to use distance from camera (Z distance) to estimate species-specific detectability.

水中ステレオ ビデオのツールを使用して魚の密度を定量化する新方式の開発

The use of video camera systems in ecological studies of fish continues to gain traction as a viable, non-extractive method of measuring fish lengths and ...

Methods for Image-based Surveys of Benthic Macroinvertebrates and Their Habitat Exemplified by the Drop Camera Survey for the Atlantic Sea Scallop

Underwater imaging has long been used in the field of marine ecology but decreasing costs of high-resolution cameras and data storage have made the approach more practical than in the past. Image-based surveys allow for initial samples to be revisited and are non-invasive compared to traditional survey methods that typically involve nets or dredges. Protocols for image-based surveys can vary greatly but should be driven by target species behavior and survey objectives. To demonstrate this, we describe our most recent methods for an Atlantic sea scallop (Placopecten magellanicus) drop camera survey to provide a procedural example and representative results. The procedure is divided into three critical steps that include survey design, data collection, and data products. The influence of scallop behavior and the survey goal of providing an independent assessment of the U.S. sea scallop resource on the survey procedure are then discussed in the context of generalizing the method. Overall, the broad applicability and flexibility of the University of Massachusetts Dartmouth School for Marine Science and Technology (SMAST) drop camera survey demonstrates the method could be generalized and applied to a variety of sessile invertebrates or habitat focused research.

Image based surveying is an increasingly practical, non-invasive method to sample the marine environment. We present the protocol of a drop camera survey that estimates the abundance and distribution of the Atlantic sea scallop (Placopecten magellanicus). We discuss how this protocol can be generalized for application to other benthic macroinvertebrates.

底生動物とその生息地大西洋帆立て貝のドロップ カメラ調査による例示のイメージ調査法

Underwater imaging has long been used in the field of marine ecology but decreasing costs of high-resolution cameras and data storage have made the ...

The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

Barnacles are marine crustaceans with a sessile adult and free-swimming, planktonic larvae. The barnacle Balanus (Amphibalanus) improvisus is particularly relevant as a model for the studies of osmoregulatory mechanisms because of its extreme tolerance to low salinity. It is also widely used as a model of settling biology, in particular in relation to antifouling research. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae for studies. A protocol for the all-year-round culturing of B. improvisus has been developed and a detailed description of all steps in the production line is outlined (i.e., the establishment of adult cultures on panels, the collection and rearing of barnacle larvae, and the administration of feed for adults and larvae). The description also provides guidance on troubleshooting and discusses critical parameters (e.g., the removal of contamination, the production of high-quality feed, the manpower needed, and the importance of high-quality seawater). Each batch from the culturing system maximally yields roughly 12,000 nauplii and can deliver four batches in a week, so up to almost 50,000 larvae per week can be produced. The method used to culture B. improvisus is, probably, to a large extent also applicable to other marine invertebrates with free-swimminglarvae. Protocols are presented for the dissection of various tissues from adults as well as the production of high-quality RNA for studies on gene expression. It is also described how cultured adults and reared cyprids can be utilized in a wide array of experimental designs for examining gene expression in relation to external factors. The use of cultured barnacles in gene expression is illustrated with studies of possible osmoregulatory roles of Na+/K+ ATPase and aquaporins.

The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

The barnacle Balanus (Amphibalanus) improvisus is a model for studying osmoregulation and antifouling. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae. Here, a protocol for the all-year-round culturing of B. improvisus is described, including the production of larvae. The use of cultured barnacles in gene expression studies is illustrated.

海洋モデルとしてフジツボタテジマフジツボ improvisus培養と遺伝子発現

Barnacles are marine crustaceans with a sessile adult and free-swimming, planktonic larvae. The barnacle Balanus (Amphibalanus) improvisus is particularly ...