The investigations were financially supported by the Division for Earth and Life Sciences of the Netherlands Organization for Scientific Research (grant 833.13.004), the Sci-Tech Landing Projection of Higher Education of Jiangxi Province (KJLD14030) and The Cultivation Plan for Young Scientists of Jiangxi Province (Jinggang star 20153BCB23014). We thank Daomeng Fu, Zhigang Li and Xiaolong Huang for their help in producing the movie.

吸引サンプリングのためのリーフブロワー-VACの1変更<ol><li>物質一覧に記載されているすべての部品を収集します。 </li><li>無可塑ポリ塩化ビニル（U-PVC）接着剤（パイプ1-2-3-4-5および6-7）で、すべてのポリ塩化ビニル（PVC）パイプを接続します。 </li><li>均等機の吸込口の周りに分布している3つの穴を開けます。 </li><li> 3つの穴のそれぞれに1本のネジを挿入することによって、塩ビ管1-5の終わり1にマシンを接続します。接続がファンをきれいに可逆的でなければならないので、マシンにパイプ端部1を接続するための接着剤を使用しないでください。 注：機械の吸込口の直径は、ここで説明したモデルと異なる場合には、パイプ端部1の直径をシームレスマシンに合うように調整されるべきです。 </li><li> 5および6を終了パイプにシールテープの2層を追加します。 </li><li>ミリアン​​ペアに吸い込まれるのサンプリングネットを防止するためにホースと口部（塩ビ管6と7）の間に金属ガーゼを入れてチャイン。 0.5ミリメートルと0.5センチメートル間のメッシュの直径を有する金網を使用してください。 </li><li>金属クランプフープと5と6を終了パイプにホースを接続します。 </li></ol> 2.サンプリングエンクロージャを準備<ol><li>プラスチックバケツ（50 L、40センチメートル底面直径）から底部を外します。筐体のこのサイズは、作物ステージ24に応じて、移植水田に2-4米の丘をカバーしています。 </li><li>ゴムバンドを使用して、バケットの上部に、1メートルの長さで、ナイロンメッシュスリーブを取り付けます。このスリーブのために、最小のターゲット節足動物の漏れを防止するために十分に小さいメッシュサイズを使用します。直径を0.5mm未満を使用してください。 </li></ol>サクションサンプリングのために変更されたリーフブロワー-VACの3分野のアプリケーション<ol><li>フィールドでデバイスを動作させるために二人を使用してください。一人は、送風機、真空を操作し、他のバケットエンクロージャとサンプリングネットを処理します。 </li><li>マシンを起動します。 </li><li>サンプリングねを挿入送風機-VACの口部へのTとゴムバンドで固定します。ネットの場合は、最小のターゲット節足動物をキャッチするのに十分小さいですが、明らかな通気抵抗を作成していないメッシュサイズを使用します。 0.2〜0.5ミリメートルの間のメッシュ径を有する光ナイロン素材を使用してください。 </li><li>フィールド内のランダムな場所に植物を介して迅速に囲いを置き、しっかりと土壌にバケツの底を押してください。スリーブは筐体の上部から逃げるの節足動物を防ぐために閉じていることを確認します。 </li><li>標準化されたサンプリング期間のために、トップダウン螺旋状の方法で、エンクロージャの内部からすべての節足動物を削除します。米の栄養における作物と生殖の段階では、それぞれ、1と2分のサンプリング期間を使用します。 </li><li>サンプルを終えた後、マシンの走行を維持しながら、ネットを閉じ、口から出て送風機-VACの一部をそれを取ると結び目とそれを閉じてすぐに、サンプリングネットからゴムバンドを削除します。 </li><li>繰り返しの次のサンプルのためのフィールド内のランダムな位置で3.2〜3.5をTEPS。複製の数は、フィールド内の節足動物の空間分布、推定値の必要な精度と研究の目的の変化に依存します。一般的に、6回の反復は、節足動物群集と種の存在量の良い印象を与えるだろう。 </li></ol>

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The authors have nothing to disclose.

吸引サンプリングは作物に節足動物コミュニティをサンプリングする多くの可能な方法の一つです。米系における科学研究のための方法は、節足動物密度の絶対的な推定値を提供するので、吸引サンプリングが適切なオプションである、それは非破壊的であり、かつ - 視覚カウントとは対照的に - 後で処理するためにサンプルの収集と保存を可能にします。市販のD-VACと比較すると、送風機、掃除機は、筐体と結合するために、小さく軽く、（浸水）水田で扱いやすいとも容易です。例えば、送風機-VACは昆虫標本のための国際標準として提示リュックD-VACモデル、一方で、体重12キロ11を有し、約6キロの重量を量ります。送風機-VACのコストは小さいながら、より重要なことには、送風機、真空のサンプリング効率は、D-VAC 16,17よりも大きいです。吸引サンプラーにリーフブロワー-VACの変更は、特別なスキルや装置を必要としませんすべての追加の部分が収集された後と1時間ほどで完了します。送風機-VACがここで説明は、文献16,17に先に説明したバージョンよりも構築・運用が容易であり、かつ必要な部品（ 表1）は、広く利用可能である標準的な建設材料です。これは、発展途上国の小さな予算で研究者にも送風機、真空にアクセスできるようになります。 エンジンのパワーと変位は、送風機、真空の吸引力を決定します。ここでは、0.7から1.2キロワットと稲植物に生息する節足動物群集をサンプリングするために十分である25-35 ccで、間の変位との間の電力を持つマシンをお勧めします。柔軟なプラスチックホースの長さと吸引口部（パイプ7）の直径は、良好なサンプリング性能のために重要です。短すぎるホースは、サンプリング中に使用する不便されるのに対し、長すぎるホースは、吸引力が低下します。同様に、小さすぎる径が小さい面によるサンプリングの効率を低下させる一方で、あまりにも大きな直径を有する口部は、吸引力を減少させます。サンプリング効率は、サンプリング期間に大きく依存します。サンプリングは、生育期間を通して行われた場合、サンプリング期間は、効率の同様のレベルを維持するために、植物の大きさ、構造、および栽植密度に調整しなければならないことがあります。サンプリング効率は、サンプリング後に囲まれた領域を注意深く目視でチェックする必要があります。まだ節足動物が存在している場合は、サンプリング時間を増加させなければなりません。栄養段階における稲作のための推奨サンプリング期間は1分であり、生殖や熟成の段階で、それは2分です。 送風機、真空で吸引サンプリングは、浸水フィールドで行うことができる、そのような落とし穴などしながら代替の方法、およびシートのサンプリングを打つ水を立っに実現可能ではありません。送風機-VACもすることができますマシンはいくつかの水を吸引することが可能であるとして、浸水水田（ 例えば 、捕食水のバグ）の水面上の節足動物群集をサンプリングするために使用されます。しかし、それは口部を水に深く挿入され、空気の流れがブロックされたときに実行を停止することができるモータとして水生節足動物をサンプリングするため推奨されません。別に米から、送風機-VACは限り植生の高さや構造がエンクロージャ25の適切な配置を可能にするように、他の作物と非作物の生息地でも使用することができます。 私たちの送風機・真空吸引サンプリング法は、非破壊的です。サンプリングネットで収集したほとんど全ての節足動物は、蚊やイトトンボなど、それらのソフトボディを含む、生き残りました。この方法の適用は、しかしながら、いくつかの制限および欠点を有します。ブロワー-VACが2人で操作する必要があります。フィールドに送風機-VACを運ぶことは、いくつかの妨害になりますので、この方法は過小評価することができますこのようなバッタなどの外乱の影響を受けやすい種。運動の前進方向に比較的乱されていない領域でのエンクロージャの高速かつ突然の配置は、この潜在的なバイアスを制限することができます。ブロア-VAC機の大きな音も障害を引き起こす可能性があり、そして住宅地における夜間のサンプリングはお勧めしません。この方法は、そのような捕食トンボ目以上蜂の寄生蜂のような移動性の高い飛翔昆虫を、サンプリングには適していません。任意のサンプリング方法と同様に、このようなスイープネットサンプリングや植物の破壊的な収穫のような他の方法、と送風機-VACの組み合わせは、節足動物群集26のより完全かつバランスのとれた評価を提供することができます。

作物における植食性や食虫性の節足動物の豊かさと多様性の繰り返し評価は、生物的防除の研究を含む、個体群動態と種間相互作用の生態研究のために必要とされます。ライスは、食虫性の節足動物1,2によって生物防除の可能性が高いと、主要な主食ですが、これは殺虫剤3によって破壊することができます。稲作における節足動物の多様性に高いこと、および節足動物の種は、様々な作物の地層（ 例えば、地面、茎、天蓋、花）を占める動き（ 例えば 、歩行、ジャンプ、飛行）のモードで異なることができると戦略を採餌（ 例えば 、固着昆虫、狩猟の捕食者と花訪問花粉媒介）4を吸引します 。 節足動物サンプリング技術、長所と短所がそれぞれの広い範囲があります。例えば、落とし穴トラップは地面住居節足動物をサンプリングするが、活動依存RELAを提供するために使用することができます的な人口は5,6を推定します 。スイープネットはキャノピー7-9に高速飛翔昆虫をサンプリングするが、節足動物の豊富さの相対的な推定値を与えるために使用することができます。ビートシート法は、植物に生息する節足動物群集をサンプリングするために使用されると節足動物の豊富さの絶対的な推定値を提供しますが、それは、このような水田10として浸水作物の分野で効果的に使用することはできませんすることができます。 フィールドの標準化された領域をカバーする筐体との組み合わせで行わ吸引サンプリングは、植物に生息する節足動物の密度の絶対的な推定値を提供します。この方法はまた、浸水イネにおいて使用することができます。サンプルは、後の処理および識別のために保存することができます。デイートリック真空（D-VAC）11は、第1 の商業的に開発された吸引サンプラーです。 D-VACSはまだ広く12-14使用されているが、それらは、比較的高価である限定された吸引力15を持っているとする彼らがハードになりますどの、比較的重いです浸水水田16で処理します。有田とHeong 16は、ガソリン駆動のリーフブロワー-VACを用いて吸引サンプラーを開発し、このプロトタイプは、さらにドミンゴとSchoenly 17により精製しました。 D-VACと比較して、送風機、真空吸引サンプラーの利点は、それがはるか​​に安く、取り扱いが容易であるということです。 ブロワー・真空吸引サンプリング方法は、多くの生態学の研究18-23で使用されてきたが、その変形およびアプリケーションのための指示が明確に記載されていません。ここでは、浸水水田における節足動物集団の吸引サンプリングのためのガソリン駆動のリーフブロワー-VACの修正とアプリケーションのビデオベース、詳細な説明を提示します。修正は、有田とHeong 16とドミンゴとSchoenly 17に触発されていますが、デザインがさらに構築および使用を容易にする、これらの元の出版物に比べて簡略化されています。

<table><tbody><tr><td colspan="4">&lt;strong&gt;Machine&lt;/strong&gt;</td></tr><tr><td>Leaf blower-vac</td><td>We used Oleo-Mac BV300, Made in Italy</td><td>Power: 1.0 kW, Displacement: 30.5 cc, Max air volume: 720 m&amp;sup3;/hr, Max air speed: 70 m/sec, Weight: 4.5 kg, Diameter of suction mouth: 113&amp;nbsp;mm</td><td>There are many different brands and models available. For comparable performance, the specifications concerning power and air speed should be similar to those presented here.</td></tr><tr><td colspan="4">&lt;strong&gt;Additional parts for modification&lt;/strong&gt;</td></tr><tr><td>PVC pipe 1</td><td></td><td>Outer &amp;oslash; of end connected to the machine: 112 mm, Inner &amp;oslash; of end connected to PVC pipe 2: 110 mm</td><td>This is the cover of a &amp;oslash; 110 mm PVC pipe</td></tr><tr><td>PVC pipe 2</td><td></td><td>Outer &amp;oslash;: 110 mm, Length: 10 cm</td><td>Normal outer &amp;oslash; 110 mm PVC pipe; to connect PVC pipe 1 and 3</td></tr><tr><td>PVC pipe 3</td><td></td><td>Inner &amp;oslash; of big end: 110 mm, Inner &amp;oslash; of small end: 50 mm</td><td>PVC &amp;oslash; 110 mm to &amp;oslash; 50&amp;nbsp;mm downpipe reducer</td></tr><tr><td>PVC pipe 4</td><td></td><td>Outer &amp;oslash;: 50 mm, Length: 5&amp;nbsp;cm</td><td>Normal &amp;oslash; 50 mm PVC pipe; to connect PVC pipe 3 and 5</td></tr><tr><td>PVC pipe 5</td><td></td><td>Inner &amp;oslash;: 50 mm and 32&amp;nbsp;mm, Outer &amp;oslash; of small part: 38 mm</td><td>PVC &amp;oslash; 50 mm to &amp;oslash; 32 mm downpipe reducer</td></tr><tr><td>Hose</td><td></td><td>Outer &amp;oslash;: 40 mm</td><td>Wire-fortified, flexible plastic hose&amp;nbsp;</td></tr><tr><td>Metal gauze</td><td></td><td>Mesh &amp;oslash;: 1 mm, &amp;oslash;: 60 mm</td><td>Prevent the sampling net from being sucked into the machine</td></tr><tr><td>PVC pipe 6</td><td></td><td>Outer &amp;oslash; of small end: 38&amp;nbsp;mm, Inner &amp;oslash; of big end: 63&amp;nbsp;mm</td><td>PVC &amp;oslash; 32 mm to &amp;oslash; 63 mm reducer</td></tr><tr><td>PVC pipe 7</td><td></td><td>Outer &amp;oslash;: 63 mm, Length: 25 cm</td><td>Normal outer &amp;oslash; 63&amp;nbsp;mm PVC pipe</td></tr><tr><td>U-PVC glue</td><td></td><td></td><td>U-PVC glue; to connect PVC parts</td></tr><tr><td>Metal clamp hoops (2)</td><td>Flexible between &amp;oslash; 35 mm - 51 mm</td><td>To connect the hose with the PVC pipes</td><td></td></tr><tr><td>Thread seal tape</td><td>Width: 18mm</td><td>Seal the hose-PVC connections</td><td></td></tr><tr><td>Screws (3)</td><td></td><td>Length: 25&amp;nbsp;mm</td><td>To connect PVC pipe 1 with the suction mouth of the machine</td></tr><tr><td colspan="4">&lt;strong&gt;Sampling net and enclosure&lt;/strong&gt;</td></tr><tr><td>Sampling net</td><td>Mesh size &amp;oslash;: 0.3 mm, Width of the mouth: 10&amp;nbsp;cm, Height: 30 cm</td><td>The sampling net has a conical shape.</td><td></td></tr><tr><td>Bucket</td><td></td><td>Bottom &amp;oslash;: 40 cm, Volume: 50 L</td><td>Cut the bottom</td></tr><tr><td>Nylon sleeve</td><td></td><td>Mesh size &amp;oslash;: 0.3 mm, Length: 1 m</td><td>To cover the bucket as enclosure</td></tr></tbody></table>

modification and application of a leaf blower-vac for field sampling of arthropods

水田は、節足動物の大きな多様性をホストするが、それらの個体群動態との相互作用を調査することは困難です。ここでは、イネの節足動物集団の吸引サンプリングのためのリーフブロワー-VACの変更、アプリケーションを記述します。筐体と組み合わせて使用​​した場合、このサンプリング装置のアプリケーションは、標準化されたサンプリング面積当たり番号などの節足動物の集団の絶対的な推定値を提供します。サンプリング効率は、サンプリング期間に大きく依存します。成熟した稲作では、0.13メートル2のエンクロージャ内の2分のサンプリングは、節足動物の人口の90％以上が得られます。デバイスはまた、水面や水田の土壌に生息する節足動物のサンプリングを可能にするが、それは、そのような捕食トンボ目以上蜂の寄生蜂のように高速飛翔昆虫を、サンプリングには適していません。修正された送風機-VACは、構築が簡単で、かつ安価と伝統的な吸引よりも扱いやすいsampliこのようなD-VACなどngのデバイス、。低コストは、発展途上国の研究者にも修正された送風機、真空にアクセスできるようになります。

<html> 295節足動物の合計が相対収量との関係を決定するために9月2015年に江西省、中国で熟成段階での稲作から8 3分ブロア-VACサンプルで収集した（節足動物の割合は、試料中の収集します）とサンプリング時間、各サンプルは、30秒毎の6つのサブサンプルに分けました。サンプルあたりの個体の平均数は（平均±SEM）36.9±4.1でした。 8節足動物注文の合計は、半翅目（28.8％）、クモ目（27.5％）および双翅目（17.6％）支配的である（ 図1）で、発見されました。相対収率は、3分後に採取節足動物の数の百分率として表し、0.9 30秒後、2および2.5分で、それぞれ（ 図2）±1.9及び97.3％で±5.1±52.9パーセント、92.4パーセントでした。 <html> 5 / 54655fig1.jpg &quot;/&gt; 江西省、中国の南昌市の郊外の節足動物のサンプルの（オーダーレベルでの） 図1 種の組成物。8サンプルは、3分ごとの送風機、掃除機で撮影しました。エラーバーは平均の標準誤差を表す。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54655/54655fig1large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図2" src="/files/ftp_upload/54655/54655fig2.jpg" /> 30秒毎のサブサンプルに基づいて、合計サンプリング時間の関数として図2の累積相対収率。3分のサンプリング期間にわたって集め節足動物の総数は、100％に設定されています。いいえ節足動物は、最後の送風機-VACのサンプルの後に目視検査の際に発見されませんでした。エラーバーは、8個の3分の試料の平均の標準誤差を表します。= &quot;https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/54655/54655fig2large.jpg&quot;ターゲット= &quot;_空白&quot;&gt;この図の拡大版をご覧になるにはこちらをクリックしてください。

Watch this Scientific Journal Video about Modification and Application of a Leaf Blower-vac for Field Sampling of Arthropods at JoVE.com

Modification and Application of a Leaf Blower-vac for Field Sampling of Arthropods

作物における節足動物の豊富さの評価は、人口動態や種間の相互作用を調査するために重要です。ここでは、イネの節足動物の吸引サンプリングのためのリーフブロワー-VACの変更、アプリケーションを記述します。

節足動物のフィールドサンプリングのためのリーフブロワー-VACの変更と応用

Rice fields host a large diversity of arthropods, but investigating their population dynamics and interactions is challenging. Here we describe the ...

modification-application-leaf-blower-vac-for-field-sampling

Universidad de Cartagena UDEC

Research

JoVE Journal

Environment

8.7K ビュー.  Wageningen University. 作物における節足動物の豊富さの評価は、人口動態や種間の相互作用を調査するために重要です。ここでは、イネの節足動物の吸引サンプリングのためのリーフブロワー-VACの変更、アプリケーションを記述します。 

ビデオ: Modification and Application of a Leaf Blower-vac for Field Sampling of Arthropods

Integrated Field Lysimetry and Porewater Sampling for Evaluation of Chemical Mobility in Soils and Established Vegetation

Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals is often not well understood. Here we demonstrate an integrated field lysimetry and porewater sampling method for evaluating the mobility of chemicals applied to soils and established vegetation. Lysimeters, open columns made of metal or plastic, are driven into bareground or vegetated soils. Porewater samplers, which are commercially available and use vacuum to collect percolating soil water, are installed at predetermined depths within the lysimeters. At prearranged times following chemical application to experimental plots, porewater is collected, and lysimeters, containing soil and vegetation, are exhumed. By analyzing chemical concentrations in the lysimeter soil, vegetation, and porewater, downward leaching rates, soil retention capacities, and plant uptake for the chemical of interest may be quantified. Because field lysimetry and porewater sampling are conducted under natural environmental conditions and with minimal soil disturbance, derived results project real-case scenarios and provide valuable information for chemical management. As chemicals are increasingly applied to land worldwide, the described techniques may be utilized to determine whether applied chemicals pose adverse effects to human health or the environment.

Field lysimetry and porewater sampling allow researchers to evaluate the fate of chemicals applied to soils and established vegetation. The goal of this protocol is to demonstrate how to install required instrumentation and collect samples for chemical analysis during integrated field lysimetry and porewater sampling experiments.

統合フィールドLysimetryと間隙水土壌中の化学物質の移動性の評価のためのサンプリングと設立植生

Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals ...

環境学

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

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

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

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

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

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

Protocols for Quantifying Transferable Pesticide Residues in Turfgrass Systems

Plant canopies in established turfgrass systems can intercept an appreciable amount of sprayed pesticides, which can be transferred through various routes onto humans. For this reason, transferable pesticide residue experiments are required for registration and re-registration by the United States Environmental Protection Agency (USEPA). Although such experiments are required, limited specificity is required pertaining to experimental approach. Experimental approaches used to assess pesticide transfer to humans including hand wiping with cotton gloves, modified California roller (moving a roller of known mass over cotton cloth) and soccer ball roll (ball wrapped with sorbent strip) over three treated turfgrass species (creeping bentgrass, hybrid bermudagrass and tall fescue maintained at 0.4, 5 and 9 cm, respectively) are presented. The modified California roller is the most extensively utilized approach to date, and is best suited for use at low mowing heights due to its reproducibility and large sampling area. The soccer ball roll is a less aggressive transfer approach; however, it mimics a very common occurrence in the most popular international sport, and has many implications for nondietary pesticide exposure from hand-to-mouth contact. Additionally, this approach may be adjusted for other athletic activities with limited modification. Hand wiping is the best approach to transfer pesticides at higher mowing heights, as roller-based approaches can lay blades over; however, it is more subjective due to more variable sampling pressure. Utility of these methods across turfgrass species is presented, and additional considerations to conduct transferable pesticide residue research in turfgrass systems are discussed.

Transferable pesticide residue experiments in turfgrass systems are integral components of human risk exposure assessments. Experimental approaches to measure transferable residues should be adjusted to the human interaction of interest and turfgrass system dynamics. Three transferable pesticide residue protocols are presented and the suitability across three turfgrass systems is discussed.

芝草システムズに譲渡残留農薬を定量化するためのプロトコル

Plant canopies in established turfgrass systems can intercept an appreciable amount of sprayed pesticides, which can be transferred through various routes ...

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 ...

Assessing the Particulate Matter Removal Abilities of Tree Leaves

Based on the conventional cleaning methods (water cleaning (WC) + brush cleaning (BC)), this study evaluated the influence of ultrasonic cleaning (UC) on collecting various sized particulate matter (PM) retained on leaf surfaces. We further characterized the retention efficiency of leaves to various sized PM, which will help to assess the abilities of urban trees to remove PM from ambient air quantitatively. 
Taking three broadleaf tree species (Ginkgo biloba, Sophora japonica, and Salix babylonica) and two needleleaf tree species (Pinus tabuliformis and Sabina chinensis) as the research objects, leaf samples were collected 4 days (short PM retention period) and 14 days (long PM retention period) after the latest rainfall. PM retained on the leaf surfaces was collected by means of WC, BC, and UC in sequence. Then, retention efficiencies of leaves (AEleaf) to three types of the various sized PM, including easily removable PM (ERP), difficult-to-remove PM (DRP), and totally removable PM (TRP), were calculated. Only around 23%-45% of the total PM retained on leaves could be cleaned off and collected by WC. When the leaves were cleaned through WC+BC, the underestimation of the PM retention capacity of different tree species was in the range of 29%-46% for various sized PM. Almost all PM retained on leaves could be removed if UC was supplemented to WC+BC. 
In conclusion, if the UC was complemented after the conventional cleaning methods, more PM on leaf surfaces could be eluted and collected. The procedure developed in this study can be used for assessing the PM removal abilities of different tree species.

The ultrasonic cleaning method was applied to elute the particulate matter (PM) retained on leaf surfaces after PM was eluted by the conventional cleaning methods (water cleaning only or water cleaning plus brush cleaning). The methodology can help to improve the estimation accuracy for PM retention capacity of leaves.

葉の木の粒子状物質の除去能力を評価します。

Based on the conventional cleaning methods (water cleaning (WC) + brush cleaning (BC)), this study evaluated the influence of ultrasonic cleaning (UC) on ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

組成と異なる環境条件下におけるバイオエアロゾルの分布解析

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...

A Method for Quantifying Foliage-Dwelling Arthropods

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density requires a method with high detection probability and a known sampling area. While most described methods provide a qualitative or semi-quantitative estimate adequate for describing species presence, richness, and diversity, few provide an adequately consistent detection probability and known or consistent sampling areas to provide an index or estimate with adequate precision to detect differences in abundance across environmental, spatial, or temporal variables. We describe how to quantify leaf-dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate and quantify them. As we demonstrate, this method can be used at a landscape scale to quantify leaf-dwelling arthropods with adequate precision to test for and describe how spatial, temporal, environmental, and ecological variables influence arthropod richness and abundance. This method allowed us to detect differences in density, richness, and diversity of leaf-dwelling arthropods among 5 genera of trees commonly found in southeastern deciduous forests.

We describe how to quantify leaf dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate for quantification.

葉住まいの節足動物を定量化する方法

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density ...

Quantifying Corticolous Arthropods Using Sticky Traps

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimate of density requires a method with high detection probability and a consistent sampling area. We used manufactured sticky traps to compare abundance, total length (a surrogate for biomass), richness, and Shannon diversity of corticolous arthropods among the boles of 5 tree species. Efficacy of this method was adequate to detect variation in corticolous arthropods among tree species and provide a standard error of the mean that was &#60;20% of the mean for all estimates with sample sizes from 7 to 15 individual trees of each species. Our results indicate, even with these moderate sample sizes, the level of precision of arthropod community metrics produced with this approach is adequate to address most ecological questions regarding temporal and spatial variation in corticolous arthropods. Results from this method differ from other quantitative approaches such as chemical knockdown, visual inspection, and funnel traps in that they provide an indication of corticolous arthropod activity over a relatively long-term, better including temporary bole residents, flying arthropods that temporarily land on the tree bole and crawling arthropods that use the tree bole as a travel route from the ground to higher forest foliage. Furthermore, we believe that commercially manufactured sticky traps provide more precise estimates and are logistically simpler than the previously described method of directly applying a sticky material to tree bark or applying a sticky material to tape or other type of backing and applying that to the tree bark.

We describe a semi-quantitative approach of measuring characteristics of corticolous (bark-dwelling) arthropod communities. We placed commercially manufactured sticky traps on tree boles to estimate abundance, total length (a surrogate to biomass), richness, and Shannon diversity for comparison among tree species.

スティッキートラップを用いたコルチコラス節足動物の定量

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimate of density ...

Low-Cost Automated Flight Intercept Trap for the Temporal Sub-Sampling of Flying Insects Attracted to Artificial Light at Night

Sampling methods are selected depending on the targeted species or the spatial and temporal requirements of the study. However, most methods for passive sampling of flying insects have a poor temporal resolution because it is time-consuming, costly and/or logistically difficult to perform. Effective sampling of flying insects attracted to artificial light at night (ALAN) requires sampling at user-defined time points (nighttime only) across well-replicated sites resulting in major time and labor-intensive survey effort or expensive automated technologies. Described here is a low-cost automated intercept trap that requires no specialist equipment or skills to construct and operate, making it a viable option for studies that require temporal sub-sampling across multiple sites. The trap can be used to address a wide range of other ecological questions that require a greater temporal and spatial scale than is feasible with previous trap technology.

To study the impacts of artificial light at night (ALAN) on nocturnal flying insects, sampling needs to be confined to nighttime. The protocol describes a low-cost automated flight intercept trap that allows researchers to sample at user-defined periods with increased replication.

夜間に人工光に引き寄せられた飛行昆虫の時間的サブサンプリングのための低コスト自動飛行傍受トラップ

Sampling methods are selected depending on the targeted species or the spatial and temporal requirements of the study. However, most methods for passive ...

Visualizing Field Data Collection Procedures of Exposure and Biomarker Assessments for the Household Air Pollution Intervention Network Trial in India

Here, we present a visual representation of standard procedures to collect population-level data on personal exposures to household air pollution (HAP) from two different study sites in a resource-constrained setting of Tamil Nadu, India. Particulate matter PM2.5 (particles smaller than 2.5 microns in aerodynamic diameter), carbon monoxide (CO), and black carbon (BC) were measured in pregnant mothers (M), other adult women (OAW), and children (C) at various times over a 4 year period. In addition, stove usage monitoring (SUMs) with data-logging thermometers and ambient measurements of air pollution were carried out. Furthermore, the feasibility of collecting biological samples (urine and dried blood spots [DBSs]) from study participants at the field sites was successfully demonstrated. Based on findings from this and earlier studies, the methods used here have enhanced the data quality and avoided issues with household air pollution and biological sample collection in resource-constrained situations. The procedures established may be a valuable educational tool and resource for researchers conducting similar air pollution and health studies in India and other low- and middle-income countries (LMICs).

We detail the consistent, high-quality procedures used throughout air and biological sampling processes at Indian field sites during a large randomized controlled trial. Insights gathered from the oversight of applications of innovative technologies, adapted for exposure assessment in rural regions, enable better field data collection practices with more reliable outcomes.

インドにおける家庭大気汚染介入ネットワーク試験のための曝露およびバイオマーカー評価のフィールドデータ収集手順の視覚化

Here, we present a visual representation of standard procedures to collect population-level data on personal exposures to household air pollution (HAP) ...