私たちは、ウォーレルウォーター・テクノロジーズ、LLCが提供する財政支援に感謝したいと思います。商標または専有物の言及は、識別のみのためのものであり、米国農務省による製品の保証または保証を意味するものではありません。

密閉室のヘッドスペース中のガス状のClO 2の1の測定<ol><li>材料を得る：のClO 2ポーチ（その放出速度（6 cm 2の総表面積）のために選択されたポリマーフィルム中のClO 2スラリー（9.5％AI）0.5gの、正確なコンポーネントが独自のもの）、ガラスチャンバー（19.14 L）、及び切り替え可能ガスの入口と出口を有する蓋。 </li><li>両面テープを使用して蓋へのClO 2ポーチを取り付けます。 </li><li>ワセリンで蓋を密封することにより、チャンバを閉じます。 </li><li>チャンバへのClO 2ガス検知器の入口と出口を接続します。 注：これはガス循環システムであり、測定を行う際に何のガス損失が発生していません。 </li><li>インキュベートした後、入口および出口ガス流をオンにし、チャンバ中のClO 2濃度を測定0、1、2、3、4、24、26、28、及び48時間。 </li><li>気性で室内の温度と相対湿度（RH）を監視atureとRHデータロガー。 </li></ol> 2.フルーツ調製および保存<ol><li>地元の小売店からの新鮮なブドウのトマト（ ナスリコペルシカム VAR。cerasiforme）の15キロを取得します。果物は健康的であり、視覚的な欠陥を持っていないことを確認してください。 </li><li> 接種材料の調製 <ol><li>接種のために柑橘類果実の表面27からの大腸菌 （野生型） およびA. alternataの株を使用します。 </li><li>その後、1日27及び再培養1日の新たなプレート上の生物の35℃での大腸菌寒天 （ECA）で培養大腸菌 。 、BAC-ループとECAプレートをサンプリングLevineのエオシンメチレンブルー（EMB）寒天上で細菌を画線し、35℃で24時間インキュベートすることによって生物を確認します。反射金属緑色を入れた培養物は、大腸菌に対して陽性です。 </li><li> 25℃でポテトデキストロース寒天（PDA）上の文化のA. alternata; Cは、胞子が表示されるまで。 </li><li>推定された濃度は、マクファーランド等価濁度標準との比較を用いて、9ログCFU / mLに達するまで滅菌蒸留水50mlに、寒天プレートから大腸菌細胞を掻き取ります。最終接種の2 Lの合計を作るために、0.1％のTween-20を含有する滅菌水1,950ミリリットルを加えます。 </li><li>エック・エイガープレート上の希釈メッキによって細胞濃度を確認してください。培養培地からのA. alternata胞子をこすり、0.1％のTween-20を含有する滅菌蒸留水2Lにそれらを停止します。 注：最終的な大腸菌集団は7.5対数CFU / gであり、 およびA. alternata集団は5.5対数CFU / gでした。 </li></ol></li><li> 完全にオートクレーブ袋で覆われている10リットルのステンレス鋼パンにトマトの7キロを置きます。安全フード内袋とパンを置きます。穏やかに攪拌しながら、上から適用トリガー噴霧器を使用して、果実に接種溶液（2L）を適用手袋をはめた手で果物。 <ol><li> 5分後、滅菌シート上の単層でトマトを配置し、それらを2時間空気乾燥することを可能にします。 twent個の1ポンド（1.14〜L）有孔クラムシェルに各果実の約200gを入れました。 </li></ol></li><li>慎重に汚染されたホイルを折り畳むとスチールパンに入れます。手袋を外して、鍋に入れます。オートクレーブ袋をラップし、25分間121℃で、すべての汚染物資をオートクレーブ。 </li><li> 12のクラムシェルの蓋にのClO 2袋を取り付けます。コントロールとして、他の12のクラムシェルを使用してください。それぞれ全体のクラムシェルを秤量します。 14日間20℃で果物を保管してください。 </li><li>一日あたりの処理ごとに、3回の反復を表す、日3、7、10でサンプルを取り、そして14サンプル3つのクラムシェル。 </li></ol>クラムシェル中のClO 2濃度の3モニタリング<ol><li>クラムシェルの中心へのClO 2ガス検知器の入口と出口チューブを挿入し、WI両端の2cmの距離目、および3日目、7、10、および14上のClO 2測定を行います。 </li></ol>微生物集団と果実品質属性の4決意<ol><li> それぞれから5つの果実（約60 g）を攪拌をオービタルシェーカー上で滅菌リン酸カリウム緩衝液（0.01M、pH7.2）中の99ミリリットルと共に滅菌サンプリングバッグで1時間、100rpmで複製します。 <ol><li>バッファー洗浄のプレート連続希釈（1-、10-、および100倍）、50μLそれぞれ、ECAにスパイラルプレーターを使用して（A. alternataのため）、PDA（E. coliについて）。 </li><li> 3日間、25℃で24時間、35℃でECAプレート及びPDAプレートをインキュベートします。光学プレートリーダーを用いて微生物のコロニー数をお読みください。使用後に汚染された果物と接触し、すべての機器をサニタイズ。 </li></ol></li><li>製造業者のプロトコルを使用して果実の硬さ試験機で果実の硬さを測定します。前にテスターを校正各使用。各複製のための20果物を測定し、1mmだけ果実を圧縮するために必要な圧力の力、ニュートン（N）として結果を表す（Nに変換・M - 1）。 </li><li>開始時および保管時のフルーツと全体のクラムシェルを秤量し、初期重量と比較して減量を計算します。 </li></ol> 5.統計解析<ol><li>三重ですべての実験を複製します。分散分析（ANOVA）を使用してデータを分析します。ダンカンの多重範囲検定による平均分離を決定します。重要性は、p &lt;0.05で定義されています。 </li></ol>

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S., Caner, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+applications+of+'active+packaging+and+chlorine+dioxide'+for+extended+shelf+life+of+fresh+strawberries.">The applications of 'active packaging and chlorine dioxide' for extended shelf life of fresh strawberries.</a> Packag Technol Sci. 24 (3), 123-136 (2011).</li><li>Paniagua, A. C., East, A. R., Hindmarsh, J. P., Heyes, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Moisture+loss+is+the+major+cause+of+firmness+change+during+postharvest+storage+of+blueberry.">Moisture loss is the major cause of firmness change during postharvest storage of blueberry.</a> Postharvest Biol Technol. 79, 13-19 (2013).</li><li>Gomez-Lopez, V. M., Ragaert, P., Jeyachchandran, V., Debevere, J., Devlieghere, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shelf-life+of+minimally+processed+lettuce+and+cabbage+treated+with+gaseous+chlorine+dioxide+and+cysteine.">Shelf-life of minimally processed lettuce and cabbage treated with gaseous chlorine dioxide and cysteine.</a> Int J Food Microbiol. 121 (1), 74-83 (2008).</li><li>Mahovic, M. J., Tenney, J. D., Bartz, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+chlorine+dioxide+gas+for+control+of+bacterial+soft+rot+in+tomatoes.">Applications of chlorine dioxide gas for control of bacterial soft rot in tomatoes.</a> Plant Dis. 91 (10), 1316-1320 (2007).</li><li>Tan, H. K., Wheeler, W. B., Wei, C. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reaction+of+chlorine+dioxide+with+amino-acids+and+peptides+-+kinetics+and+mutagenicity+studies.">Reaction of chlorine dioxide with amino-acids and peptides - kinetics and mutagenicity studies.</a> Mutat Res. 188 (4), 259-266 (1987).</li></ol>

The authors declare that they have no competing financial interests.

二酸化塩素は、食品の腐敗を防止するための理想的な殺生物剤です。しかし、高価なジェネレータ又は非効率的な二部粉末混合を必要とする、高濃度及び非輸送性で不安定です。この研究は、食品の腐敗や食中毒の発生率を低減するための二酸化塩素の安定した、すぐに使用できる形式の適用を検討しました。現在使用されている他の二酸化塩素の応用技術とは対照的に、ここで使用される?...

新鮮な果物や野菜が豊富な食事は、冠状動脈性心臓病や癌1の特定の種類を含む多くの疾患のリスクを軽減するのを助けることができます。しかし、農産物2を汚染食べる消費者の間で病気や死さえを引き起こす可能性があり、新鮮な果物や野菜の消費に伴う、 大腸菌 、 サルモネラ菌 、およびリステリア菌などの食中毒微生物病原体の数が、あります。例えば、 大腸菌 O157：H7の発生は4、ブドウ、トマト、イチゴ及び3に関連している、およびA型肝炎の発生は、新鮮なブルーベリー5に関連しています。また、微生物汚染は収穫後の崩壊6を通じてかなりの製品損失を引き起こす可能性があります。 ルナリアのalternataは重要な植物病原性真菌トンであります<html>帽子は、植物7の380以上の宿主種における葉のスポットや他の病気を引き起こすことが知られています。 ルナリア黒点8、幹潰瘍病とトマト9の葉枯病の原因であることが示されています。したがって、安全で効果的な収穫後の汚染除去処理の両方の制御食品媒介病原体に必要とされ、新鮮な農産物で収穫後腐敗を防止します。 低・非剰余技術は、代替消毒剤のための新しいトレンドです。収穫後の殺菌剤の様々な腐敗生物を低減し、食中毒を予防するために使用されています。オゾン、強力な抗菌剤は、イチゴとブルーベリー10、11の品質と鮮度を保つことが示されています。しかし、オゾンは、果実の表面組織の酸化を引き起こし、変色や風味品質の劣化につながることができますS = &quot;外部参照&quot;&gt; 12。塩素は、ブルーベリーやリンゴ13のように、生鮮食品を消毒するために使用されています。有効ながら、塩素は、新鮮な果物15の消毒のために使用する場合は特に、発癌性副生成物14が得られ、窒素含有化合物又はアンモニアと反応することができます。 二酸化塩素（二酸化塩素2）、消毒剤の代替は、果物や野菜16の収穫後の治療のために、中国と米国の両方によって承認されました。 ClO 2は、遊離塩素17のそれよりも2.5倍高い酸化能を有する水溶性酸化剤です。 ClO 2は非常に低濃度で効果的で短い接触時間18です。 ClO 2は、消毒のために使用される濃度で毒性が低く、最小限の腐食性を有し、それは最も効果的な殺菌の一つとして認識されていますそして、設定19、20、21の様々な使用のための殺菌剤。 数多くの研究結果は、のClO 2は、食品媒介病原体と収穫後の腐敗16を制御することができることを示しています。 H7及びブルーベリーとイチゴ腐敗22,23を防止するために：例えば、のClO 2ガスは、 リステリア・モノサイトゲネス 、 サルモネラ 、 大腸菌 O157を不活性化するために使用されてきました。新鮮なフルーツの属性を維持しながらのClO 2ガスは、微生物汚染のリスクを低減し、そしてそれはイチゴ24の収穫後の減衰を制御するのに有効でした。しかし、歴史的サイトまたは非効率的な二部粉末混合に高価な発電機を必要とする、高濃度及び非輸送性で不安定です。 しかし、新しいのClO既製、制御放出製剤（ すなわち、それは発電機または成分の予備混合を必要としない）で2製品が予備実験25において食品腐敗生物及び病原体を制御するのに非常に有効であることが示されました。それは、安全で費用対効果の高い、非腐食性、容易に輸送、および環境に対する悪影響はないとのClO 2の制御放出形態です。以前の実験は、濾過材で包み、クラムシェル包装に入れ、この徐放性のClO 2粉末は著しく、新鮮なブルーベリーとイチゴの崩壊を減少ベリー水分損失を減少し、収穫後の記憶部25、26間に果実硬度を維持することを実証しました。最近では、制御放出のClO 2のパケットは、半透性ポリマー膜中のClO 2のスラリー状の封止によって開発されました。この作業の目的でした：1）両方の密閉容器中のClO 2つのガス放出特性を監視し、穿孔クラムシェルで、2）のClO 2ポーチは食品媒介病原体に対する容器及びグレープトマトの減衰で囲まれた制御放出の効果を調べ、そして3）グレープトマトのストレージ・品質上の制御放出のClO 2の効果を評価します。

<table><tbody><tr><td>Curoxin&lt;sup&gt;&amp;#174;&lt;/sup&gt; chlorine dioxide pouch</td><td>Worrell Water Technologies</td><td></td><td>Slurry, a.i. 9.5% in sealed semi-permeable polymer film</td></tr><tr><td>Grape tomato</td><td>Santa Sweets, Inc</td><td></td><td>Santa Sweets Authentic&amp;nbsp;</td></tr><tr><td>ClO&lt;sub&gt;2&lt;/sub&gt; gas detector</td><td>Analytical Technology, Inc., Collegeville, PA</td><td></td><td>PortaSens II&amp;nbsp;</td></tr><tr><td>Perforated clamshell</td><td>Packaging Plus LLC, Yakima, WA</td><td></td><td>OSU #1, 1 lb</td></tr><tr><td>&lt;em&gt;Escherichia coli&lt;/em&gt;&amp;nbsp;</td><td></td><td></td><td>Wild Type (WT) from fruit surface</td></tr><tr><td>Alternaria alternata</td><td></td><td></td><td>from fruit surface</td></tr><tr><td>&lt;em&gt;E. coli&lt;/em&gt; agar&amp;nbsp;</td><td>EC Broth, Oxoid, UK</td><td></td><td>EC Broth with 1.5% agar</td></tr><tr><td>Potato dextrose agar&amp;nbsp;</td><td>BD Difco, Sparks, MD</td><td></td><td></td></tr><tr><td>Levine eosin methylene blue agar</td><td>BD Difco, Sparks, MD</td><td></td><td></td></tr><tr><td>Trigger spray bottle&amp;nbsp;</td><td>Impact Products, LLC., Toledo, OH</td><td></td><td></td></tr><tr><td>Sterilized sampling bag&amp;nbsp;</td><td>Fisherbrand, Fisher Scientific, Pittsburgh, PA</td><td></td><td></td></tr><tr><td>Orbit shaker&amp;nbsp;</td><td>New Brunswick Scientific, New Brunswick, NJ</td><td></td><td>Innova 2100</td></tr><tr><td>IUL Instruments Neutec Eddy jet spiral plater inoculation plating system</td><td>Neutec Group Inc., Farmingdale, NY</td><td></td><td></td></tr><tr><td>EZ micro optical plate reader&amp;nbsp;</td><td>Synoptics, Ltd., Cambridge, UK</td><td></td><td>ProtoCOL</td></tr><tr><td>Fruit firmness tester&amp;nbsp;</td><td>Bioworks Inc, Wamego, KS</td><td></td><td>FirmTech 2&amp;nbsp;</td></tr><tr><td>Tinytag temperature and RH data logger</td><td>Gemini Data Loggers, West Sussex, UK</td><td></td><td></td></tr><tr><td>McFarland equivalence turbidity standard</td><td>Fisherbrand, Fisher Scientific, Pittsburgh, PA</td><td></td><td></td></tr></tbody></table>

controlled-release of chlorine dioxide in a perforated packaging system to extend the storage life and improve the safety of grape tomatoes

制御放出二酸化塩素（のClO 2）パウチ半透性ポリマーフィルムへのClO 2のスラリー状の封止によって開発されました。ポーチの放出特性を持つか、果物のない容器にモニターしました。パウチはグレープトマトを含む穿孔クラムシェルの内部に固定し、微生物集団、堅さ、および体重減少に対する効果は20°Cで14日間の保存期間中に評価しました。 3日以内に、クラムシェル中のClO 2濃度は3.5 ppmのに達し、その後10日目まで一定のままで、それは/のClO 2パウチは3.08対数CFUによって大腸菌の個体数を減少させる、強力な抗菌活性を示し、14日目によっては2ppmまで低下しました貯蔵の14日後の2.85対数CFU / GによってG及びアルテルナリアalternata集団 。 ClO 2処理は、軟化および体重減少を低減し、トマトの全体的な貯蔵寿命を延長しました。我々の結果ClO 2治療は貯蔵寿命を延長し、その品質を損なうことなく保存中のトマトの微生物の安全性を向上させるために有用であることを示唆しています。

ClO 2のリリースでは、最初の数時間にわたる直線状のパターンを示しました。濃度は、最初の4時間にわたって約2.38 ppmの/ hに増加しました。放出速度は、インキュベーションの24時間後に鈍化、及びのClO 2濃度は25.4 ppmで達しました。しかし、濃度はインキュベーションの24時間（ 図1）の後に安定である傾向がありました。 <p class="jove_c...

Watch this Scientific Journal Video about Controlled-release of Chlorine Dioxide in a Perforated Packaging System to Extend the Storage Life and Improve the Safety of Grape Tomatoes at JoVE.com

Controlled-release of Chlorine Dioxide in a Perforated Packaging System to Extend the Storage Life and Improve the Safety of Grape Tomatoes

Here, we describe a protocol for the application of a novel, slow-release ClO2 product that reduces spoilage and extends the shelf life of fresh fruit. The slow-release ClO2 product was added to standard commercial grape tomato packaging and tested against Escherichia coli and Alternaria alternata.

ストレージ寿命を延長し、ブドウトマトの安全性を向上させるための穴があいた包装システムにおける二酸化塩素の放出制御

A controlled-release chlorine dioxide (ClO2) pouch was developed by sealing a slurry form of ClO2 into semipermeable polymer film; the release properties ...

controlled-release-chlorine-dioxide-perforated-packaging-system-to

Universidad de Cartagena UDEC

Research

JoVE Journal

Environment

10.9K ビュー.  USDA, ARS, USHRL. Here, we describe a protocol for the application of a novel, slow-release ClO2 product that reduces spoilage and extends the shelf life of fresh fruit. The slow-release ClO2 product was added to standard commercial grape tomato packaging and tested against Escherichia coli and Alternaria alternata.

ビデオ: Controlled-release of Chlorine Dioxide in a Perforated Packaging System to Extend the Storage Life and Improve the Safety of Grape Tomatoes

Utilizing the Ethylene-releasing Compound, 2-Chloroethylphosphonic Acid, as a Tool to Study Ethylene Response in Bacteria

Ethylene (C2H4) is a gaseous phytohormone that is involved in numerous aspects of plant development, playing a dominant role in senescence and fruit ripening. Exogenous ethylene applied during early plant development triggers the triple response phenotype; a shorter and thicker hypocotyl with an exaggerated apical hook. Despite the intimate relationship between plants and bacteria, the effect of exogenous ethylene on bacteria has been greatly overlooked. This is partly due to the difficulty of controlling gaseous ethylene within the laboratory without specialized equipment. 2-Chloroethylphosphonic acid (CEPA) is a compound that decomposes into ethylene, chlorine, and phosphate in a 1:1:1:1 molar ratio when dissolved in an aqueous medium of pH 3.5 or greater. Here we describe the use of CEPA to produce in situ ethylene for the investigation of ethylene response in bacteria using the fruit-associated, cellulose-producing bacterium Komagataeibacter xylinus as a model organism. The protocols described herein include both the verification of ethylene production from CEPA via the Arabidopsis thaliana triple response assay and the effects of exogenous ethylene on K. xylinus cellulose production, pellicle properties and colonial morphology. These protocols can be adapted to examine the effect of ethylene on other microbes using appropriate growth media and phenotype analyses. The use of CEPA provides researchers with a simple and efficient alternative to pure ethylene gas for the routine determination of bacterial ethylene response.

The protocols outlined herein facilitate the convenient investigation of bacterial ethylene responses by utilizing 2-chloroethylphosphonic acid (CEPA). Ethylene is produced in situ through the decomposition of CEPA in an aqueous bacterial growth medium, circumventing the requirement for pure ethylene gas.

細菌でエチレン応答を研究するためのツールとして、エチレン放出化合物、2-クロロエチルホスホン酸を利用します

Ethylene (C2H4) is a gaseous phytohormone that is involved in numerous aspects of plant development, playing a dominant role in senescence and fruit ...

生化学

The Portable Chemical Sterilizer (PCS), D-FENS, and D-FEND ALL: Novel Chlorine Dioxide Decontamination Technologies for the Military

There is a stated Army need for a field-portable, non-steam sterilizer technology that can be used by Forward Surgical Teams, Dental Companies, Veterinary Service Support Detachments, Combat Support Hospitals, and Area Medical Laboratories to sterilize surgical instruments and to sterilize pathological specimens prior to disposal in operating rooms, emergency treatment areas, and intensive care units. The following ensemble of novel, &#8216;clean and green&#8217; chlorine dioxide technologies are versatile and flexible to adapt to meet a number of critical military needs for decontamination6,15. Specifically, the Portable Chemical Sterilizer (PCS) was invented to meet urgent battlefield needs and close critical capability gaps for energy-independence, lightweight portability, rapid mobility, and rugged durability in high intensity forward deployments3. As a revolutionary technological breakthrough in surgical sterilization technology, the PCS is a Modern Field Autoclave that relies on on-site, point-of-use, at-will generation of chlorine dioxide instead of steam. Two (2) PCS units sterilize 4 surgical trays in 1 hr, which is the equivalent throughput of one large steam autoclave (nicknamed &#8220;Bertha&#8221; in deployments because of its cumbersome size, bulky dimensions, and weight). However, the PCS operates using 100% less electricity (0 vs. 9 kW) and 98% less water (10 vs. 640 oz.), significantly reduces weight by 95% (20 vs. 450 lbs, a 4-man lift) and cube by 96% (2.1 vs. 60.2 ft3), and virtually eliminates the difficult challenges in forward deployments of repairs and maintaining reliable operation, lifting and transporting, and electrical power required for steam autoclaves.

The Portable Chemical Sterilizer PCS, D-FENS, and D-FEND ALL: Novel Chlorine Dioxide Decontamination Technologies for the Military

The Portable Chemical Sterilizer (PCS) is a revolutionary, energy-independent, almost waterless sterilization technology for Army medical units. The PCS generates chlorine dioxide from dry reagents mixed with water on-site, at-will, and at point-of-use (PoU) in a plastic suitcase. The Disinfectant-sprayer for Foods and ENvironmentally-friendly Sanitation (D-FENS) and the Disinfectant for ENvironmentally-friendly Decontamination, All-purpose (D-FEND ALL) produce aqueous chlorine dioxide in a collapsible spray bottle and other potential embodiments. These versatile decontamination technologies kill microbes in myriad diverse Dual-use applications for military and civilian consumers.

ポータブル化学殺菌（PCS）、D-FENSおよびD-FEND ALL：小説二酸化塩素除染技術の軍事用

There is a stated Army need for a field-portable, non-steam sterilizer technology that can be used by Forward Surgical Teams, Dental Companies, Veterinary ...

生物工学

Procedures of Laboratory Fumigation for Pest Control with Nitric Oxide Gas

Nitric oxide (NO) is a newly discovered fumigant for postharvest pest control. This paper provides detailed protocols for conducting NO fumigation on fresh products and procedures for residue analysis and product quality evaluation. An airtight fumigation chamber containing fresh fruit and vegetables is first flushed with nitrogen (N2) to establish an ultralow oxygen (ULO) environment followed by injection of NO. The fumigation chamber is then kept at a low temperature of 2 - 5 &#176;C for a specified time period necessary to kill a target pest to complete a fumigation treatment. At the end of a fumigation treatment, the fumigation chamber is flushed with N2 to dilute NO prior to opening the chamber to ambient air to prevent the reaction between NO and O2, which produces NO2 and may damage delicate fresh products. At different times after NO fumigation, NO2 in headspace and nitrate and nitrite in liquid samples were measured as residues. Product quality was evaluated after 2 weeks of post-treatment cold storage to determine effects of NO fumigation on product quality. Keeping&#160;O2 from reacting with NO is critical to NO fumigation and is an important part of the protocols. Measuring NO levels is challenging and a practical solution is provided. Possible protocol modifications are also suggested for measuring NO levels in the fumigation chambers as well as residues. NO fumigation has the potential to be a practical alternative to methyl bromide fumigation for postharvest pest control on fresh and stored products. This publication is intended to assist other researchers in conducting NO fumigation research for postharvest pest control and accelerating the development of NO fumigation for practical applications.

This paper describes nitric oxide (NO) fumigation protocols for postharvest pest control. Fumigation chambers are flushed with nitrogen (N2) to establish ultralow oxygen conditions before NO is injected. At the end, chambers are flushed with N2 to dilute NO before exposing products to ambient air to prevent exposure to NO2.

窒素酸化物ガスで害虫駆除のため研究所くん蒸処理の手順

Nitric oxide (NO) is a newly discovered fumigant for postharvest pest control. This paper provides detailed protocols for conducting NO fumigation on ...

環境学

Culturing and Maintaining Clostridium difficile in an Anaerobic Environment

Clostridium difficile is a Gram-positive, anaerobic, sporogenic bacterium that is primarily responsible for antibiotic associated diarrhea (AAD) and is a significant nosocomial pathogen. C. difficile is notoriously difficult to isolate and cultivate and is extremely sensitive to even low levels of oxygen in the environment. Here, methods for isolating C. difficile from fecal samples and subsequently culturing C. difficile for preparation of glycerol stocks for long-term storage are presented. Techniques for preparing and enumerating spore stocks in the laboratory for a variety of downstream applications including microscopy and animal studies are also described. These techniques necessitate an anaerobic chamber, which maintains a consistent anaerobic environment to ensure proper conditions for optimal C. difficile growth. We provide protocols for transferring materials in and out of the chamber without causing significant oxygen contamination along with suggestions for regular maintenance required to sustain the appropriate anaerobic environment for efficient and consistent C. difficile cultivation.

Culturing and Maintaining Clostridium difficile in an Anaerobic Environment

Clostridium difficile is a pathogenic bacterium that is a strict anaerobe and causes antibiotic associated diarrhea (AAD). Here, methods for isolating, culturing and maintaining C. difficile vegetative cells and spores are described. These techniques necessitate an anaerobic chamber, which requires regular maintenance to ensure proper conditions for optimal C. difficile cultivation.

培養およびメンテナンス<em&gt;クロストリジウム·ディフィシル</em&gt;嫌気性環境での

Clostridium  difficile is a Gram-positive, anaerobic, sporogenic bacterium  that is primarily responsible for antibiotic associated diarrhea (AAD) and is ...

免疫・感染学

An Efficient Clearing Protocol for the Study of Seed Development in Tomato (Solanum lycopersicum L.)

Tomato (Solanum lycopersicum L.) is one of the major cash crops worldwide. The tomato seed is an important model for studying genetics and developmental biology during plant reproduction. Visualization of finer embryonic structure within a tomato seed is often hampered by seed coat mucilage, multi-cell-layered integument, and a thick-walled endosperm, which needs to be resolved by laborious embedding-sectioning. A simpler alternative is to employ tissue clearing techniques that turn the seed almost transparent using chemical agents. Although conventional clearing procedures allow deep insight into smaller seeds with a thinner seed coat, clearing tomato seeds continues to be technically challenging, especially in the late developmental stages.
Presented here is a rapid and labor-saving clearing protocol to observe tomato seed development from 3 to 23 days after flowering when embryonic morphology is nearly complete. This method combines chloral hydrate-based clearing solution widely used in Arabidopsis with other modifications, including the omission of Formalin-Aceto-Alcohol (FAA) fixation, the addition of sodium hypochlorite treatment of seeds, removal of the softened seed coat mucilage, and washing and vacuum treatment. This method can be applied for efficient clearing of tomato seeds at different developmental stages and is useful in full monitoring of the developmental process of mutant seeds with good spatial resolution. This clearing protocol may also be applied to deep imaging of other commercially important species in the Solanaceae.

An Efficient Clearing Protocol for the Study of Seed Development in Tomato Solanum lycopersicum L.

The tomato seed is an important model for studying genetics and developmental biology during plant reproduction. This protocol is useful for clearing tomato seeds at different developmental stages to observe the finer embryonic structure.

トマトの種子発育の研究のための効率的な清算プロトコル(Solanum lycopersicum L.)

Tomato (Solanum lycopersicum L.) is one of the major cash crops worldwide. The tomato seed is an important model for studying genetics and developmental ...

発生生物学

High-Throughput Identification of Resistance to Pseudomonas syringae pv. Tomato in Tomato using Seedling Flood Assay

Tomato is an agronomically important crop that can be infected by Pseudomonas syringae, a Gram-negative bacterium, resulting in bacterial speck disease. The tomato-P. syringae pv. tomato pathosystem is widely used to dissect the genetic basis of plant innate responses and disease resistance. While disease was successfully managed for many decades through the introduction of the Pto/Prf gene cluster from Solanum pimpinellifolium into cultivated tomato, race 1 strains of P. syringae have evolved to overcome resistance conferred by the Pto/Prf gene cluster and occur worldwide.
Wild tomato species are important reservoirs of natural diversity in pathogen recognition, because they evolved in diverse environments with different pathogen pressures. In typical screens for disease resistance in wild tomato, adult plants are used, which can limit the number of plants that can be screened due to their extended growth time and greater growth space requirements. We developed a method to screen 10-day-old tomato seedlings for resistance, which minimizes plant growth time and growth chamber space, allows a rapid turnover of plants, and allows large sample sizes to be tested. Seedling outcomes of survival or death can be treated as discrete phenotypes or on a resistance scale defined by amount of new growth in surviving seedlings after flooding. This method has been optimized to screen 10-day-old tomato seedlings for resistance to two P. syringae strains and can easily be adapted to other P. syringae strains.

High-Throughput Identification of Resistance to Pseudomonas syringae pv. Tomato in Tomato using Seedling Flood Assay

The seedling flood assay facilitates rapid screening of wild tomato accessions for the resistance to the Pseudomonas syringae bacterium. This assay, used in conjunction with the seedling bacterial growth assay, can assist in further characterizing the underlying resistance to the bacterium, and can be used to screen mapping populations to determine the genetic basis of resistance.

シュードモナスシリンゲpvに対する耐性のハイスループット同定苗フラッドアッセイを用いたトマトのトマト

Tomato is an agronomically important crop that can be infected by Pseudomonas syringae, a Gram-negative bacterium, resulting in bacterial speck disease. ...

Isolation of Native Soil Microorganisms with Potential for Breaking Down Biodegradable Plastic Mulch Films Used in Agriculture

Fungi native to agricultural soils that colonized commercially available biodegradable mulch (BDM) films were isolated and assessed for potential to degrade plastics. Typically, when formulations of plastics are known and a source of the feedstock is available, powdered plastic can be suspended in agar-based media and degradation determined by visualization of clearing zones. However, this approach poorly mimics in situ degradation of BDMs. First, BDMs are not dispersed as small particles throughout the soil matrix. Secondly, BDMs are not sold commercially as pure polymers, but rather as films containing additives (e.g. fillers, plasticizers and dyes) that may affect microbial growth. The procedures described herein were used for isolates acquired from soil-buried mulch films. Fungal isolates acquired from excavated BDMs were tested individually for growth on pieces of new, disinfested BDMs laid atop defined medium containing no carbon source except agar. Isolates that grew on BDMs were further tested in liquid medium where BDMs were the sole added carbon source. After approximately ten weeks, fungal colonization and BDM degradation were assessed by scanning electron microscopy. Isolates were identified via analysis of ribosomal RNA gene sequences. This report describes methods for fungal isolation, but bacteria also were isolated using these methods by substituting media appropriate for bacteria. Our methodology should prove useful for studies investigating breakdown of intact plastic films or products for which plastic feedstocks are either unknown or not available. However our approach does not provide a quantitative method for comparing rates of BDM degradation.

Plastic films labeled "biodegradable" are commercially available for agricultural use as mulches. Tillage represents an attractive disposal method, but degradation under field conditions is poorly understood. The purpose of this study was to develop methods for isolating native soil fungi and bacteria that colonize plastic mulch films after field burial.

農業で使用する生分解性プラスチックマルチフィルムを分解するための可能性を持つネイティブ土壌微生物の分離

Fungi native to agricultural soils that colonized commercially available biodegradable mulch (BDM) films were isolated and assessed for potential to ...

生物学

Standardized Method for High-throughput Sterilization of Arabidopsis Seeds

Arabidopsis thaliana (Arabidopsis) seedlings often need to be grown on sterile media. This requires prior seed sterilization to prevent the growth of microbial contaminants present on the seed surface. Currently, Arabidopsis seeds are sterilized using two distinct sterilization techniques in conditions that differ slightly between labs and have not been standardized, often resulting in only partially effective sterilization or in excessive seed mortality. Most of these methods are also not easily scalable to a large number of seed lines of diverse genotypes. As technologies for high-throughput analysis of Arabidopsis continue to proliferate, standardized techniques for sterilizing large numbers of seeds of different genotypes are becoming essential for conducting these types of experiments. The response of a number of Arabidopsis lines to two different sterilization techniques was evaluated based on seed germination rate and the level of seed contamination with microbes and other pathogens. The treatments included different concentrations of sterilizing agents and times of exposure, combined to determine optimal conditions for Arabidopsis seed sterilization. Optimized protocols have been developed for two different sterilization methods: bleach (liquid-phase) and chlorine (Cl2) gas (vapor-phase), both resulting in high seed germination rates and minimal microbial contamination. The utility of these protocols was illustrated through the testing of both wild type and mutant seeds with a range of germination potentials. Our results show that seeds can be effectively sterilized using either method without excessive seed mortality, although detrimental effects of sterilization were observed for seeds with lower than optimal germination potential. In addition, an equation was developed to enable researchers to apply the standardized chlorine gas sterilization conditions to airtight containers of different sizes. The protocols described here allow easy, efficient, and inexpensive seed sterilization for a large number of Arabidopsis lines.

The objective of this study was to determine the effects of bleach and chlorine gas sterilization on seed germination of a range of Arabidopsis genotypes grown on sterile media. Optimized sterilization protocols have been developed to prevent the growth of microbial contaminants while providing satisfactory seed survival.

高スループット シロイヌナズナ種子の殺菌のための標準化された方法

Arabidopsis thaliana (Arabidopsis) seedlings often need to be grown on sterile media. This requires prior seed sterilization to prevent the growth of ...

直接注入装置を用いた植物への分子送達

Direct Infusion Device for Molecule Delivery in Plants

Testing the function of therapeutic compounds in plants is an important component of agricultural research. Foliar and soil-drench methods are routine but have drawbacks, including variable uptake and the environmental breakdown of tested molecules. Trunk injection of trees is well-established, but most methods for this require expensive, proprietary equipment. To screen various treatments for Huanglongbing, a simple, low-cost method to deliver these compounds to the vascular tissue of small greenhouse-grown citrus trees infected with the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas) or infested with the phloem-feeding CLas insect vector Diaphorina citri Kuwayama (D. citri) is needed.
To meet these screening requirements, a direct plant infusion (DPI) device was designed that connects to the plant&#39;s trunk. The device is made using a nylon-based 3D-printing system and easily obtainable auxiliary components. The compound uptake efficacy of this device was tested in citrus plants using the fluorescent marker 5,6-carboxyfluorescein-diacetate. Uniform compound distribution of the marker throughout the plants was routinely observed.
Furthermore, this device was used to deliver antimicrobial and insecticidal molecules to determine their effects on CLas and D. citri respectively. The aminoglycoside antibiotic streptomycin was delivered into CLas-infected citrus plants using the device, which resulted in a reduction in the CLas titer from 2 weeks to 4 weeks post treatment. Delivering the neonicotinoid insecticide imidacloprid into D. citri-infested citrus plants resulted in a significant increase in psyllid mortality after 7 days. These results suggest that this DPI device represents a useful system for delivering molecules into plants for&#160;testing and facilitate research and screening purposes.

著者スポットライト:植物における分子送達のための直接注入装置  

This manuscript describes a novel direct plant infusion device for screening the effectiveness of molecules against the bacteria (Candidatus Liberibacter asiaticus) or its insect vector (Diaphorina citri, Kuwayama) that, in combination, are associated with Huanglongbing citrus disease.

植物における分子送達のための直接注入装置

Testing the function of therapeutic compounds in plants is an important component of agricultural research. Foliar and soil-drench methods are routine but ...

ストレージ寿命を延長し、ブドウトマトの安全性を向上させるための穴があいた包装システムにおける二酸化塩素の放出制御

要約

さらに動画を探す

この動画の章

細菌でエチレン応答を研究するためのツールとして、エチレン放出化合物、2-クロロエチルホスホン酸を利用します

ポータブル化学殺菌（PCS）、D-FENSおよびD-FEND ALL：小説二酸化塩素除染技術の軍事用

窒素酸化物ガスで害虫駆除のため研究所くん蒸処理の手順

培養およびメンテナンス

トマトの種子発育の研究のための効率的な清算プロトコル(Solanum lycopersicum L.)

シュードモナスシリンゲpvに対する耐性のハイスループット同定苗フラッドアッセイを用いたトマトのトマト

農業で使用する生分解性プラスチックマルチフィルムを分解するための可能性を持つネイティブ土壌微生物の分離

高スループットシロイヌナズナ種子の殺菌のための標準化された方法

植物における分子送達のための直接注入装置

植物における分子送達のための直接注入装置

ストレージ寿命を延長し、ブドウトマトの安全性を向上させるための穴があいた包装システムにおける二酸化塩素の放出制御

要約

さらに動画を探す

この動画の章

細菌でエチレン応答を研究するためのツールとして、エチレン放出化合物、2-クロロエチルホスホン酸を利用します

ポータブル化学殺菌（PCS）、D-FENSおよびD-FEND ALL：小説二酸化塩素除染技術の軍事用

窒素酸化物ガスで害虫駆除のため研究所くん蒸処理の手順

培養およびメンテナンス

トマトの種子発育の研究のための効率的な清算プロトコル(Solanum lycopersicum L.)

シュードモナスシリンゲpvに対する耐性のハイスループット同定苗フラッドアッセイを用いたトマトのトマト

農業で使用する生分解性プラスチックマルチフィルムを分解するための可能性を持つネイティブ土壌微生物の分離

高スループット シロイヌナズナ種子の殺菌のための標準化された方法

植物における分子送達のための直接注入装置

植物における分子送達のための直接注入装置

高スループットシロイヌナズナ種子の殺菌のための標準化された方法